Qatar Foundation Annual Research Conference Proceedings Volume 2014 Issue 1

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

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

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

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

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

Diabetes is highly prevalent in Qatar and about 1/3 of patients are not aware of their diseases. Screening for undiagnosed diabetes is essential for effective management and prevention of diabetes and its complications. The effectiveness and cost analysis of several diabetes screening programs have been the subject of intensive investigation. Point of care (POC) measurement of capillary blood glucose (CBG) is very simple and can be applied widely, however, random CBG values are hard to interpret unless a high cut-off point is utilized, such as >200 mg/dL. A high cut off point would miss a large number of diabetics and pre-diabetics. POC CBG measurement of fasting and post-prandial values is difficult to organize in a community sitting. In Qatar, and all Muslim countries, most people observe fasting. If POC CBG is applied between 12:00 and sunset, then all values are at least 9 h after the morning meal and are considered fasting values. Similarly, POC CBG measured after evening prayer (Taraweeh) is equivalent to a late post-prandial value (~3 h after evening meal). Here we piloted a study to evaluate the usefulness of POC CBG to screen for diabetes in Ramadan after Juma'a prayer (~9 h of fast) and after Taraweeh prayer (3-4 h after evening meal - Iftar) in the large Mosq in Doha. We also administered a questionnaire to learn about known diabetes, co-morbidities and family history. A total of 2177 individuals were screened in 2 days, 75% were men and 25% were women representative of 40 different nationalities with most from Egypt (743, 38%), India (348, 23%) and Qatar (146, 7%). The distribution of CBG values were not statistically different between afternoon and evening values and were pooled for this analysis. 57% of all values were normal (<100 mg/dL). 27% had pre-diabetic CBG values (100-124 mg/dL). The remaining, 17% had diabetic values, 12% were previously known diabetic and 5% can be considered as newly diagnosed diabetes. Analysis of age, family history, ethnic background, comorbidities revealed the following facts: known DM were older, had more comorbidities (hypertension, kidney and heart diseases, and smoking). Presence of at least one diabetic parent existed in 37% of normal individual and 53% of known diabetics. Presence of two DM parents existed in only 8% of normal individual and 21% of known diabetics. There was a significant correlation between body weight and CBG values across the whole cohort when known diabetic values are excluded. In conclusion, this pilot study shows that Ramadan is an opportunity for screening for diabetes that can be applied on a wide community efficiently. *This study was financially supported by Action on Diabetes.

Extracellular signal regulated kinase (ERK1/2) is a member of the mitogen-activated protein kinase pathway (MAPK). ERK1/2 has a wide variety of functions including cell proliferation, differentiation, and migration to name a few. Thus alteration in the ERK1/2 pathway can result in different pathologies such as cancer, cardiovascular diseases and diabetes. ERK MAP kinase is activated through binding of extracellular growth factors such as PDGF and EGF to their respective receptors. Calreticulin (CRT) is an endoplasmic reticulum (ER) chaperone that aids in the protein folding and maturation. It also plays a role in the ER quality control process. Our lab previously illustrated that CRT knockout cells compensate for ER stress by activating ER associated protein degradation. We also demonstrated that loss of CRT increased cellular resistance to apoptosis due to activation of Akt pathway. Activation of ERK signaling pathway was shown to protect against ER-induced cell death. To date little is known about the role of CRT in the ERK pathway. Thus the objective of our study was to examine changes in ERK1/2 kinase and the growth factor receptors in CRT knockout cells. Western blot analysis with anti-ERK1/2 antibody illustrated no significant change in the ERK1/2 protein level in crt−/− as compared to wt cells. However, the basal p-ERK1/2 was significantly higher in the crt−/− as compared to wt cells after overnight starvation. Interestingly, FBS stimulation resulted in increased phosphorylation of ERK1/2 to the same level in both cell lines. On the other hand 10 min EGF stimulation resulted in a significantly higher p-ERK1/2 in CRT knockout cells as compared to the wt cells. Overall, these data illustrate that loss of CRT function results in increased ERK phosphorylation and might contribute to the observed increased resistance to apoptosis and cell proliferation.

Functional annotation of genes and their protein products is an essential step in the course of genome analysis. Experimental functional analysis techniques such as microarray or yeast two-hybrid systems simply can not handle the quantity of sequences made available by next-generation sequence technologies, and thus annotation of gene products is primarily predicted applying computational tools. A variety of computational methods are now available applying different methodologies, amongst others, homology-, sequence-, structure- or network-based methods. Nonetheless, so far there is no method that predicts the function of a group of genes and their products; for instance genes that are expressed during the course of a disease or cellular stress. We developed a computational pipeline that fuzes different network data sources, namely protein-protein interaction, gene ontology, phylogenetic, gene expression and pathway information, in order to predict the group function(s) of genes. The main steps of the pipeline are, first, the network integration of the different input sources, second, the clustering of the involved genes according to their similarity, and, third, the (re-)assignment of genes/proteins with unknown function. These steps are repeated until the algorithm converges into one or more final clusters/groups, which are additionally mapped onto KEGG pathways in order to biologically identify and interpret higher-level systemic/organismic functions. We successfully applied the pipeline to different groups of genes over-expressed in diseases of major interest in Qatar, such as type-2 diabetes, breast cancer and pancreatic cancer.

Mesenchymal stem cells (MSC) are self-renewing multipotent cells which hold great potential in reconstructive medicine and tissue engineering. They have the ability to differentiate into cells of the mesoderm lineage and have been shown to be beneficial for the treatment of a variety of diseases. MSC can be derived from multiple adult tissues but have only limited expansion capacity in cell culture. Highly proliferative ESC-derived MSC can be an alternative source for MSC but currently no standardized protocol exists which meets clinical standards. We further developed and improved a protocol (Raynaud et al., 2013) to differentiate human embryonic stem cells (ESC) into highly-proliferative MSC. ESC-derived MSCs were tested for their characteristic surface markers by flow cytometry and the differentiation capabilities typical for MSCs (bone, fat) were verified. To characterize the differentiation process in-depth we performed comprehensive large-scale proteomic and phosphoproteomic profiling experiments using quantitative high resolution mass spectrometric analysis based on reductive dimethylation (Boersema et al., 2009). Experiments were designed as triplex comprising an internal standard, labeled with light isotopes, and time points, labeled either with medium or heavy isotopes. Differentiation was followed over a time course of 30 days including sampling days 0, 1, 2, 5, 15, and 30. Samples for proteomic analysis were fractionated by in-solution isoelectric focusing prior to mass spectrometry. Phosphoproteomic profiling was performed according to the TiSH protocol (Engholm-Keller et al., 2012) including phosphopeptide enrichment by titanium dioxide combined with sequential immobilized metal affinity chromatography. ESC-derived MSC were compared to adult tissue-derived MSC (bone marrow MSC) as well as to their origin (ESC). In total, 8615 proteins were identified with 5800 proteins on average quantified per sample. A total of 4064 proteins were quantified in all samples at all stages and were subjected to stringent statistical analysis. For phosphoproteomics, we identified and quantified more than 8000 phosphosites on 4000 proteins with around 4600 phosphosites on 1800 proteins per sample. A large overlap (75%) was observed for quantified proteins between proteomic and phosphoproteomic workflows. To further enhance the analytical depth, data was integrated with transctriptome profiling derived from next-generation RNA sequencing which enabled us to quantify the expression of over 14000 genes. To identify important regulators of differentiation, we performed clustering according to expression patterns. The established differentiation protocol is highly reproducible and robust, may be adapted for clinical use and thus be of great value for the stem cell research community. The comprehensive analysis of the differentiation process will improve understanding of MSC biology and therefore directly benefit MSC-based therapies. References Boersema, P.J., et al. (2009). Multiplex peptide stable isotope dimethyl labeling for quantitative proteomics. Nat Protoc. 4, 484-494. Engholm-Keller, K., et al. (2012). TiSH--a robust and sensitive global phosphoproteomics strategy employing a combination of TiO2, SIMAC, and HILIC. J. Proteomics 75, 5749-5761. Raynaud, C.M., et al. (2013). Human Embryonic Stem Cell Derived Mesenchymal Progenitors Express Cardiac Markers but Do Not Form Contractile Cardiomyocytes. PLoS ONE 8.

Introduction and objective: Adhesion can be defined as fibrous bands of scar-like tissue that appear between two surfaces inside the body. Tissue adhesion formation is one of the common problems in postoperative lower abdomen or uterus in reproductive organs; it can lead to severe complications such as pain and infertility. The objective of this study was to evaluate the effect of standardised extract of Eurycoma longifolia (SEEL) in reducing adhesion formation in the uterus of estradiol valerate (EV)-treated rats induced by coitus. Method: Adult (12-14 weeks-old) female rats with normal Oestrous cycle (OC) were randomly divided into two groups (n = 8 for each group). Each rat (in both groups) was given of EV 2.0 mg/rat. One month after the EV injection, each rat in Group 1 was given SEEL at a dose of 50 mg/kg/d for four weeks. In Group 2, distilled water was orally administered to each rat. After 14 days of SEEL or distilled water treatment, each rat in both groups was allowed to mate with a proven fertile male rat. The duration of the mating was 10-14 days. Results: In the group treated with EV+ SEEL plus mating with a fertile male, the animals looked healthy when compared to the EV+mating group. However, neither implantation nor foetuses were observed during the macroscopical examination in either group. Adhesion on the uterus was not noted in the EV+ SEEL + mating animals. Adhesion formation in the EV+ SEEL+ mating animals was significantly reduced by 12.5% compared with that in the EV+mating group, which had 75%. Conclusion: the present study demonstrates that TAF 273 has been found to show a reduction in adhesion formation.

About 70% of human breast cancers express and are dependent for growth on estrogen receptor α (ERα), and therefore are sensitive to antiestrogen therapies. However, progression to an advanced more aggressive phenotype is associated with acquisition of resistance to antiestrogens and/or invasive potential. In the present study, we highlight the role of the serine/threonine-protein kinase D1 (PKD1) in ERα-positive breast cancers. Growth of ERα-positive MCF-7 and MDA-MB-415 human breast cancer cells was assayed in adherent or anchorage-independent conditions in cells overexpressing or depleted for PKD1. PKD1 induces cell growth through both an ERα-dependent manner, by increasing ERα expression and cell sensitivity to 17β­estradiol, and an ERα-independent manner, by reducing cell dependence to estrogens and conferring partial resistance to antiestrogen ICI 182,780. PKD1 knockdown in MDA-MB-415 cells strongly reduced estrogen-dependent and independent invasion. Quantification of PKD1 mRNA levels in 38 cancerous and non-cancerous breast cell lines and in 152 ERα-positive breast tumors from patients treated with adjuvant tamoxifen showed an association between PKD1 and ERα expression in 76.3% (29/38) of the breast cell lines tested and a strong correlation between PKD1 expression and invasiveness (p<0.0001). In tamoxifen-treated patients, tumors with high PKD1 mRNA levels (n=77, 50.66%) were significantly associated with less metastasis-free survival than tumors with low PKD1 mRNA expression (n=75, 49.34%) (p=0.031). Moreover, PKD1 mRNA levels are strongly positively associated with EGFR and vimentin levels (p<0.0000001). Thus, our study defines PKD1 as a novel attractive prognostic factor and a potential therapeutic target in breast cancer.

I. BACKGROUND & O BJECTIVES Recent advances in sensing, communication and actuation are leading to the next generation of Telemedicine when integrated with Wireless Body Area Networks (WBANs). They have a great potential in fostering the provision of next-generation Ubiquitous Healthcare (U-Health). However, deploying new technologies in healthcare applications without considering security makes patient privacy vulnerable, especially when we deal with highly sensitive physiological data of a patient such as heart-rate, position, temperature etc. Because traditional security mechanisms have been designed for the systems with sufficient resources, they cannot be applied directly to the extremely resource constrained WBANs devices. Our main objective is to provide a lightweight security mechanism suitable for WBANs and propose a fully functional secure healthcare platform to monitor remotely the patients' health status. II. METHODS The proposed healthcare system (Fig. 1) integrates heterogeneous devices and wearable medical sensors. It informs the healthcare professionals by sending Patient Health Information (PHI) from Body Sensors to Body Router (BR). BR uses wireless local area network to communicate with Gateway in indoor environment, and it communicates with Server through 3G/4G in outdoor environment. Gateway and Server request Hardware Address Resolution Process (HARP) for security. To guarantee the system security, we propose (i) a new lightweight encryption scheme based on stream cipher, where the encryption key is changed after each round of transmission to provide strong confidentiality (ii) an authentication mechanism is provided through HARP to authenticate BRs with their unique ID assigned before deployment and to guarantee that only authorized healthcare professionals can access to patients' data thanks to their biometrical information. III. RESULTS In order to investigate the feasibility of the proposed secure healthcare system, all components have been implemented in an experimental testbed. Zolertia is used as a wearable BS, which is a MSP430 microcontroller based sensor node equipped with CC2420 RF transceiver for wireless communication. Cubox is used as a BR, which is a low power ARM architecture. PCs are used as Gateway/Server and HARP. Firstly, the security tests of the encryption scheme have been provided to check the randomness quality. The results were significantly higher (from 0.342178 to 0.974321) than the indicated threshold value (0.01) recommended by National Institute of Standards and Technology (NIST). Secondly, the end-to-end delay was around 97ms with encryption scheme and 94ms without encryption. Thus, the communication overhead incurred due to the encryption algorithm for each round is nearly 3ms. IV. CONCLUSION Experimental results have shown that our U-Health solution is feasible in real-world scenarios. The proposed encryption scheme passed all randomness tests recommended by NIST and the security functionality brings only 3ms communication overhead in terms of end-to-end delay per transmission.

Combined clinical, neuroimaging, genomics, and molecular genetics research efforts between neuropediatrics clinic at HMC, international clinical collaborators, and neurogentics lab at WCMCQ has successfully enabled a prominent achievement in providing tools and defined strategy for diagnostics, and primary prevention, carrier detection and prenatal intervention, of a wide variety of neurogenetics diseases encountered in Gulf and Arab patients. Whole genome sequencing, Sanger sequencing, in-vitro studies were our main experimental tools to prove the gene defect and to derive the well characterized diagnostic tool. Examples of diseases that are identified in our collection of families and now possible to screen and diagnose on gene basis include AR-Leukodystrophy with subcortical cysts [VDK], AR-LGMD-sarcoglycans, AR-HSP-related genes, AR-hyperexplexia and other interesting neurogentics disorders. The presentation will show a summary of clinical, neuroimaging, novel genes/ mutations and the preventive measures that have been actually undertaken in those diseases.

We develop a mobile application for Community Health Workers (CHWs) to collect health data for monitoring children's growth and development with a custom mobile application. The application is designed to be delay tolerant and optimized for low-resource settings. The evaluation is carried out in an urban and a rural location in Rwanda. The preliminary results show that CHWs are very successful in electronic data collection for tasks they already routinely perform, such as measuring weight gain and Middle Upper Arm Circumference (MUAC), which is a UNICEF protocol for detecting malnutrition. They had no problem adjusting to the electronic system since we followed the same format as the paper forms. We consider how mobile data collection improves both data availability and accuracy. The considerations that can relate to the advantages of electronic data versus the present paper-based system are: timeliness, accuracy and consistency. There is no doubt that electronic data is timelier than paper reports since the present system are submitted once a month. For data accuracy, we compare the accuracy of the present paper system with the Electronic Health Record approach. Our application has a built in error correction because it issues a warning when data is outside of the normal range of ±2sd. It does not prevent CHW to enter the data but it warns them of a possibility of an error, and improves the internal validity of data. By comparing the values that are input with a model based on previous measurements, it is possible to ensure self-consistency and detect errors. We have data from the paper records for the six months before we started the study at the same two locations. We were not able to locate and follow the same children using both systems because the paper data is fragmented and incomplete. However, we have many records available and the quantity of data will facilitate comparison of the two systems and estimates of statistical significance of the results. In addition we build a distribution model based on a large data set from the Rwandan Ministry of Health concerning the weight of children under five. We test how the electronic and paper data is described by this distribution over successive measurements and use the results to develop improved methods for checking the accuracy of data input by CHW in real-time.

Increasing evidence suggests that the c-Abl protein tyrosine kinase could play a role in the pathogenesis of Parkinson's disease (PD) and other neurodegenerative disorders. c-Abl has been shown to regulate the degradation of two proteins implicated in the pathogenesis of PD, parkin and alpha-synuclein (alpha-syn). The inhibition of parkin's neuroprotective functions is regulated by c-Abl-mediated phosphorylation of parkin. However, the molecular mechanisms by which c-Abl activity regulates -syn toxicity and clearance remain unknown. Herein, using NMR spectroscopy, mass spectrometry, in vitro enzymatic assays and cell-based studies, we established that alpha-syn is a bona fide substrate for c-Abl. In vitro studies demonstrate that c-Abl directly interacts with alpha-syn and catalyzes its phosphorylation mainly on tyrosine 39 (pY39), and to a lesser extent on tyrosine Y125 (pY125). Analysis of human brain tissues showed for the first time that pY39 alpha-syn is detected in the brains of healthy individuals and those with PD. Interestingly, whereas Nilotinib, a specific inhibitor of c-Abl kinase activity, induces alpha-syn protein degradation via the autophagy and proteasome pathways, the overexpression of alpha-syn in the midbrain of rats enhances c-Abl expression. Together, these results suggest that changes in c-Abl expression and/or activation play important roles in regulating alpha-syn clearance and contribute to the pathogenesis of PD. Thus, targeting c-Abl kinase activity represents a promising therapeutic strategy for the treatment of PD and related synucleinopathies. This work was funded by ERC, NIH (#R37-AG019391 and #R24AA012725) and the Swiss cancer league (KLS-3132-02-2013, KLS-3132-02-2013). Our work was recently published in Hum Mol Genet. 2014 Jun 1;23(11):2858-79.

Background and Objectives Impaired cholesterol and fat metabolism contributes to obesity, type 2 diabetes and atherogenic cardiovascular disease; major chronic conditions that are increasingly prevalent in Qatar. There is thus an urgent need for new treatment modalities to combat the rise in these diseases. Understanding how cholesterol/lipid homeostasis is achieved and maintained is among the very first critical steps towards developing new strategies for better treatments. Despite the intricate underlying mechanisms, many regulatory factors have been found to be involved in the metabolic regulation of lipids. Recent studies found that microRNAs (miR), short 22-nucleotide RNA molecules that control gene expression by silencing target mRNAs at the translational level, hold promise as therapeutic targets since they contribute to the etiology of many pathological conditions. We found that microRNA 33a (miR-33a) embedded within the intronic sequence of SREBP-2 gene, the master regulator of cholesterol metabolism, acts as a key modulator of intracellular cholesterol trafficking pathways. Here, we report our new findings on the 5p and 3p strands of miR-33a (miR-33a-5p and miR-33a-3p) in regulating cholesterol homeostasis. Methods Using computational algorithms we identified potential target genes of both strands of miR-33a that are involved in cholesterol metabolism. We then used luciferase reporter assays, wherein the 3'-UTR of the target gene was fused to a luciferase reporter gene, to validate the predicted targets of miR-33a. Positive associations between the microRNAs and their targets were further confirmed by mutating the binding sites on the target gene 3'-UTR and performing luciferase assays with the mutant constructs. Overexpression and knockdown of the microRNAs in human liver cell lines were carried out with mimics and inhibitors, respectively, and their effect on target gene expression was studied by RT-qPCR and western blotting. Cholesterol uptake and efflux assays in hepatocytes were exploited to better assess the functional roles of miR-33a. Results Intriguingly, we found that in addition to our pervious discovery of miR-33a-5p reducing cholesterol efflux from hepatocytes and macrophages, miR-33a-3p promotes intracellular cholesterol uptake. While miR-33a-5p inhibits ABCA1, a cholesterol efflux pump instrumental in raising plasma HDL-cholesterol levels and reverse cholesterol transport, miR-33a-3p activates LDL-receptor (LDLR) by repressing both Idol and PCSK9, two major negative regulators of LDLR. Accordingly, in an SREBP-2 active condition antisense-mediated inhibition of miR-33a-3p, but not miR-33a-5p, led to decreased LDLR expression and LDL uptake in human hepatocytes. Conclusions Mutations in the LDLR gene are well known to be highly associated with atherosclerosis and familial hypercholesterolemia. Our data unravels a novel regulatory circuit by which LDLR, which is transcriptionally activated by SREBP-2, is further protected from degradation at the post-translational level by miR-33a-3p. This indicates that both the 5p and 3p strands of miR-33a are mutually exclusive in elevating intracellular cholesterol levels along with their SREBP-2 host gene. These findings provide impetus for further characterization and development of new therapeutic interventions in cardiovascular disease.

Thymidylate kinases (TMKs) play a central role in the production of nucleotide precursors that are required for the replication of DNA. Consequently, this enzyme is a potential drug target for the discovery of anti-bacterial, anti-fungal, and anti-parasitic drugs. In addition, TMKs are also involved in the activation of prodrugs. In particular, the anti-HIV drug AZT is activated by human TMK (huTMK) and the low efficiency of huTMK towards AZT is a significant problem in the use of AZT in the treatment of HIV. Finally, nucleotide precursors are required in large amounts by cancerous cells, thus the inhibition of huTMK by chemotherapeutic agents may enhanced the arsenal of drugs that are used to treat cancer. Although there has been some effort to develop inhibitors of TMKs, these efforts have been hampered by the difficulty in performing high throughput screening using compound libraries. In addition, the characterization of TMK-drug complexes has been limited to X-ray diffraction studies which provide static information about the enzyme-drug complex. There have been no attempts to apply high-resolution multi-nuclear NMR techniques to determine the fundamental dynamic properties of these enzymes and how the structure and dynamics of the enzyme are altered by the binding of substrates or inhibitors. As a preliminary step in characterizing these enzymes by NMR we have over-expressed TMKs from yeast, human, and two pathogens - Plasmodium falciparum and Candida albicans. Expression of these TMKs was optimized by the design of synthetic genes for expression in bacteria. In the case of the human enzyme, we are able to routinely produce 250 mg of the enzyme/L of culture. Preliminary NMR spectra of the yeast, human, and plasmodium enzyme show that the protein is a homo-dimer in solution, as anticipated from X-ray studies. The amide and methyl spectra are well resolved, indicating that resonance assignment by traditional TROSY based methods will be feasible for both the amides and the methyl resonances. In particular the high sensitivity and dispersion of the methyl spectra will facilitate characterization of the dynamic properties of these enzymes by carbon and deuterium relaxation. Ligand induced changes in the dynamics and structure of huTMK in solution will be characterized using NMR methods. These studies will provide additional insights into the inability to huTMK to effectively activate AZT. The entropic component of the thermodynamics of substrate binding to TMK from the parasite that causes malaria will also be characterized by determining dynamic changes by NMR methods. The development of NMR methods to study these enzymes also provides a method for high throughput screening of compound libraries by detecting chemical shift changes in the NMR spectral of the enzyme due to binding of a potential lead compound.

Identification of post-translationally modified α-Synuclein protein in biofluids of Parkinson's disease patients using a targeted and quantitative mass spectrometry approach. Céline Vocat1, Bruno Fauvet1, Michel Prudent4, Adrien W. Schmid3, Hilal A. Lashuel1&2. 1 Laboratory of Molecular and Chemical Biology of Neurodegeneration, Brain Mind Institute, Ecole Polytechnique Fédérale de Lausanne (EPFL), 1015 Lausanne, Switzerland. 2. Qatar Biomedical Research Institute, 5825 Doha, Qatar. 3. Proteomics Core Facility, Ecole Polytechnique Fédérale Lausanne (EPFL), Switzerland. 4. Service Régional Vaudois de Transfusion Sanguine, Unité de Recherche et Développement, Switzerland. Parkinson's disease (PD) is a movement disorder characterized by the progressive loss of dopaminergic neurons and the presence of intracellular protein inclusions (Lewy Bodies) found in the brain of affected patients. Protein aggregation and post-translational modifications (PTMs), such as the site specific phosphorylation of alpha-Synuclein (α-Syn) protein have been reported to be strongly linked to PD pathogenesis. Therefore, pathologically modified α-Syn species represent a primary target for the diagnosis and treatment of PD. In this work, we aimed at conducting a comprehensive study, using multiple mass spectrometry and proteomics based approaches, to assess the chemical heterogeneity of α-Syn and to identify and map the pattern of α-Syn PTMs in plasma and red blood cells from PD and dementia with Lewy bodies (DLB) patients compared to healthy, age-matched control subjects. More specifically, we focused on the pattern of PTMs in the blood in order to identify if these modifications correlate with α-Syn PTM's observed in the brain and cerebrospinal fluid (CSF) during disease progression. The use of full-length, heavy isotope-labelled (15N) α-Syn protein and peptide standards with site-specific modifications, which mirror the key pathological PTMs of α-Syn found in PD, with targeted proteomics and selected reaction monitoring (SRM) mass spectrometry have enabled us to specifically identify and monitor single or multiple site-specific phosphorylations, N-terminal acetylation, truncations and splice variants of α-Syn. We have developed a multiplexed SRM assay which allows us to monitor several PTMs during a single analytical run. The identification of a specific isoform or PTMs pattern that correlate with PD or DLB could provide novel insights into the mechanism of the disease development, contribute to the identification of novel therapeutic targets and most importantly, could provide a diagnostic marker to detect and monitor the progression of PD and related synucleinopathies.

It is well established that intermittent exercises are very specific for performance in field and combat sports. However, few studies have examined the effect of sinusoidal oscillation in exercise intensity could maintain or ameliorates energetic coast. The aim of this work was to investigate if the variation of exercise allures (constant speed (CT-sp) vs. sinusoidal speed (SIN-sp) on physiological responses during submaximal exercise. Ten male footballers (182.6 ± 6.2 cm and 79.6 ± 6.4 kg) were volunteered to participate to this study. After measuring maximal aerobic velocity (MAV) and corresponding maximal oxygen uptake (VO2max) during the University of Montreal incremental test', subjects performed, in a randomized order, six test sessions of 10 min at different intensities (65, 75 and 85% VMA) in either CT-sp (a constant distance of 12.5 m between cones) or SIN-sp with an amplitude of 3 km.h-1 (alternating distances of 9.85 m and 15.15 m in each speed). Heart rate (HR), blood lactate concentration [La] and oxygen uptake (VO2) were determined during each test session. Results showed that HR, [La] and VO2 were higher during SIN-sp than CT-sp in the different exercise intensities. In addition, multiple linear regression was performed as below: Y = a *X1+ b * X2+ C with X2 corresponding to exercise allure (EA) (Vcte vs Vsin) as independent variable (taking the value of 0 or 1 ) to study the relationships: VO2 / VO2max = a * HR / HR max + b * EA + C, [La] / [Lamax] = a * HR / HR max + b * TE + C and [La] / [Lamax ] VO2 = a * / b * VO2max + TE + C. The statistical analysis shows that only the VO2/ VO2max and HR / HRmax were significant (P <0.001). The results of this study raise the question of the effectiveness of sinusoidal training allure on cardiorespiratory and metabolic parameters in football players.

Background & Objective: Cerebral aneurysms are one of the prevalent and devastating cerebrovascular diseases of adult population worldwide. The resulting effect is subarachnoid hemorrhage, intra- cerebral hematoma and other complications leading to a high mortality rate. When the aneurysm is fusiform, having wide neck or is large in shape, deploying stent in the parent artery to bypass aneurysm is considered as the most suitable treatment. The stent graft is designed to seal tightly with your artery above and below the aneurysm. The graft is stronger than the weakened artery and it allows your blood to pass through it without pushing on the bulge. So that blood cannot flow through the aneurysm to cause any future complication including rupture. Therefore, separation of parent artery from the aneurysm is immensely desired. This paper presents a method to separate parent artery from the aneurysm. Method: It has been challenging to distinguish the parent artery from the aneurysm geometry using a computer algorithm [1]. To date, only a few approaches to accomplish this task have been proposed. In our method, an initial surface mesh of the parent artery with aneurysm is first generated. Then the following steps are subsequently performed to separate the artery from the aneurysm. Step 1. The user specifies foreground and background on the mesh by placing centerline (Figure (a)) on the parent artery and aneurysm; it is also useful for generating Voronoi diagram (Figure (b) and (c)). Step 2. A feature preserving harmonic field based on the user specification is generated (Figure (d)). The resultant harmonic (i.e., "intensity") field over the artery geometry contains large variations not only at these concave and high curvature regions but also at the borders between the normal parent artery and the aneurysm. Since the parent artery centerline is nominally influenced by the presence of an aneurysm, the parent centerline is reconstructed; deviation is recorded and used while finalizing the average isoline or cutting boundary. Step 4. The isolines are generated with the help of Voronoi diagram (from which the average isoline is extracted); see Figure (e). We consider the isolines of the resultant harmonic field as the potential cutting boundary of the parent artery from the aneurysm. Along an isoline, the field variation is minimum. Step 5. A graph-based technique [2] is applied on the harmonic field to segment the parent artery from the aneurysm utilizing the average isoline and distance metric, where we define the energy function according to the harmonic field on the mesh. Results & Conclusion: For testing the method, we collected CTA slices with average thickness of 0.29mm, pixel spacing of 0.29mm x 0.29mm, and matrix size 512x512 on five subjects at the Hamad Medical Corporation using the Siemens Axiom Artis Interventional suite. The average time required by MATLAB R14 to perform segmentation is 2 m for one subject by a 2 GB RAM and core2duo processor (without optimization). Experimental results have shown satisfactory results for meshes with either simple or complicated model.

Background: Diabetic nephropathy (DN) is a chronic and serious complication associated with diabetes. The standardization of an in vitro model to best represent DN is very challenging due to the chronic nature of the condition. Therefore, two different renal tubule cell lines - Madin-Darby canine kidney cells (MDCK) and Normal rat kidney cells (NRK-52E) - were used to investigate the effects of high glucose on kidney cells. Objective: To determine the effects of high glucose concentrations on cell viability (using MTT assay), oxidative stress (using dichlorofluorescein (DCF) staining), and expression of proteins activated in DN such as aldose reductase and glucose-regulated protein-78 (GRP78), an endoplasmic reticulum chaperone (using western blotting). Results: MDCK cells showed a subtle decrease in viability when exposed to high glucose concentrations (30 mM and 1% FBS) for 48 h. Furthermore, there was a slight increase in aldose reductase expression after 48 h of high glucose exposure, however; the GRP78 levels remained unchanged. NRK-52E cells showed more consistent decrease in viability after 48 and 72 h of high glucose exposure (30 mM and 1% FBS). In addition, the DCF staining also demonstrated an increase in oxidative stress after 24 h of high glucose exposure. Furthermore, a 30% increase in aldose reductase expression has been observed after 48 h of high glucose exposure. Conclusion: Although the 48 h high glucose exposure in MDCK cells can be used as a model for in vitro DN, the results are less reproducible, whereas NRK-52E cells seem to be a better and more reliable cell line to mimic the features of DN in vitro. Key words: Diabetic nephropathy; In vitro; Kidney; Oxidative Stress; ER Stress.

Background. Insulin resistance and the prevalence of diabetes are high in the Middle Eastern population and in people of South Asian origin. Because of epidemic proportions that diseases have reached in these populations. It is important to find early markers along with preventative interventions. Recent data indicates that the metabolic defect in the pre-diabetic condition relates more strongly to post-prandial deficiency than to the fasted state. Women have lower levels of risk factors for metabolic disease when assessed in the post-absorptive state. However, very few reports have investigated these in the post-prandial state, especially amongst an Arab population. Objectives. This study investigated systemic cardiovascular risk factors both in the fasted and post-prandial state in a healthy, non-diabetic female local population. Methods. A cohort of young female, non-diabetic subjects representative of a general Qatari population was recruited. The study was approved by the national ethical committee and all subjects gave written informed consent. Subjects attended after an overnight fast and blood samples were taken prior to and 30 and 120 minutes after ingesting a liquid mixed meal. Anthropometric measures included age, height, weight, blood pressure and pulse. Bioimpedance was used to measure body fat (%, mass and distribution) andBasel Metabolic Rate {BMR},(Tanita MC-980). Glucose (hexokinase, Roche), lipids (Roche), insulin and proinsulin (Mercodia),Glucose like peptide1 {GLP-1},(Millipore) and adipokines (R & D Systems) were all determined. Data were analysed by SPSS version 22.0 for windows. Parametric tests were used for normally distributed data and non-parametric analysis for skewed dataare shown in the text as Mean {SD} or Median {Interquertal range }. Results. The subjects were young (Age 29.8 {4.9} years), non-obese (BMI 25.6 {4.7} kg/m2}, normotensive (systolic BP 109 {10} and diastolic BP 72 {6} mmHg) and normolipidaemic (Total-cholesterol 3.5 {0.7}, LDL-cholesterol 2.0 {0.4}, HDL-cholesterol 1.3 {0.3}, triglycerides 0.6 {0.2} mmol/L). Their total body fat was relatively high (34 {6.5} %), which was reflected by elevated levels of systemic leptin (30.8 {17.5-53.0} ng/ml) and lower adiponectin (8.1 {6.0-11.3} ?g/ml). Despite no apparent fasting or post-prandial hyperglycaemic and HOMA-IR levels being normal, post-prandial hyperinsulinaemia and hyperproinsulinaemia were significant (see Table). Proinsulin also constituted 19% of total insulin-like molecules. Discussion. Young normal-weight Qatari women with no apparent metabolic disease are hyperinsulinaemic and hyperproinsulinaemic in the fed state. Thus, elevated post-prandial levels of insulin-like molecules may be an early, sensitive marker for the metabolic defect that precedes cardiometabolic disease in this population. Table. Fasting and post-prandial concentrations of insulin-like molecules VariablesFasting30 minutes120 minutes Glucose (mmol/L)4.6 (0.3)5.0 (0.7)4.3 (0.5) Insulin (mIU/L)5.1 (4.0-6.3)50.5 (31.5-57.9)28.8 (23.2-37.4) Proinsulin (pmol/L)8.1 (5.7-11.4)22.3 (14.0-38.7)34.3 (22.0-49.6) GLP-1 (pmol/L)2.0 (1.8-2.4)8.4 (6.0-11.3)7.0 (3.3-9.6) Data ar shown as mean (SD) or median (interquartile range).

ABSTRACT Introduction Over the last 30 years the incidences of cancer and diabetes have been increasing progressively and there is both epidemiologic and experimental data linking diabetes and various cancer outcomes. Previous studies has shown that physical activity, height, and obesity; measured by weight, waist circumference, waist-to-hip ratio and body mass index were associated with the risk of diabetes and common cancer outcomes such as breast, colorectal and prostate cancer. However, there is little data on whether these modifiable risk factors are predictive of these malignancies among diabetics. The identification of factors that modify the risk of cancer among diabetics could lead to better surveillance or intervention to reduce cancer incidence among those at highest risk. Aim The aim of this study was to observe the relationship between anthropometric measures, including weight, height, and adiposity and physical activity with the risk of breast, colorectal and prostate cancer among diabetic individuals within the European Prospective Investigation into Cancer and Nutrition cohort. Methods Data from 27,365 diabetics from nine European countries between the ages of 20 to 85 years and a mean follow-up of 11.1 years from the European Prospective Investigation into Cancer and Nutrition (EPIC) was used. Of the 27,365 diabetics, 546 developed breast cancer, 363 developed prostate cancer and 308 developed colorectal cancer. Hazard ratios (HR) and 95% confidence interval (CI) were estimated using a Cox proportional hazard model, stratified by age in one year increments, gender and centre and adjusted for smoking status, alcohol consumption, education, BMI and physical activity. The HR was used to examine the association between anthropometric measures at recruitment and PA estimated from questionnaires with breast, prostate and colorectal cancer. Results There were no significant associations between height, weight, waist circumference waist-to-hip ratio, BMI and physical activity with breast, prostate and colorectal cancer when comparing the highest and lowest quartiles. However, there was a significant association between height and breast cancer when comparing the third quartile to the first (Q3 vs Q1, HR: 3.34, 95%CI: 1.09-10.22). Moreover, there was a suggestive inverse association between physical activity with breast and prostate cancer (P-trend 0.085 and P-trend 0.07, respectively). There was also a suggestive positive association between abdominal obesity and colorectal cancer (P-trend 0.04 for waist circumference and P-trend 0.08 for waist-to-hip ratio). Conclusion In this study of diabetic patients nested within the EPIC cohort, there was little evidence for an association between anthropometric measures and physical activity with breast, prostate and colorectal cancer. However, there was a suggestive association between physical activity with breast and prostate cancer and a suggestive association between abdominal obesity and colorectal cancer. Due to the limited number of cases in this study, further investigations between the associations of these modifiable factors with these cancers among diabetics are required.

This project focuses on developing a complete telemetry system in response to the growing demand for efficient, mobile and inexpensive system for detecting cardiovascular abnormalities. Our team has already developed two algorithms. The first algorithm detects various critical points on Electrocardiograph (ECG) waveforms such as: P-wave, QRS complex, T-wave and ST elevation. Based on these points, the second algorithm detects Myocardial Infractions (MI) in a patient. Currently, we are developing a telemetry system that comprises of an Arduino microcontroller and a smartphone. The 12-lead ECG signals are collected from a patient or an ECG simulator. The signals are then sent to an electronic circuit for amplification and filtration. The amplified and filtered signals are collected in the Arduino microcontroller. The microcontroller sends the signals, via Bluetooth, to an Android smartphone application developed by our team. The ECG data is sent from the smartphone to a webserver using 3G or Wi-Fi connection. The data is securely processed and analyzed in the webserver using the algorithms developed by our team. The processed ECG waveforms and the results of its analysis are sent back and displayed on the smartphone application. The analyzed results display whether the patient has a likelihood of having an MI. Furthermore, the analyzed results are stored in a secure database for future reference. Using the real-time analyzed ECG waveforms, the integrated system is designed to provide early warning of cardiac risk, thus saving valuable time and effort in patients' treatment. Along with that, this system will be efficient in terms of cyber security, cost management and reliability.

Background: Breast Cancer is one of the leading causes of cancer related mortality in women, both in Qatar and the world. Despite the available treatments the incidence of breast cancer is increasing. This highlights the need for new approaches for cancer. One of the fields that is gaining attention nowadays is herbal medicine. Herbs are known to have bioactive compounds that affect many diseases one of which is cancer. Origanum syriacum is an herb that is frequently used in Mediterranean region. Recently, it has been established that O. syriacum possess anti-proliferative activity in non-invasive breast cancer. Although it has some medicinal values, it remains poorly investigated. Here we tested the anti-tumor activity of O. syriacum extract (OSE) on the aggressive human breast cancer cell line, MDA-MB-231. Methods: The extract was prepared by dissolving the leaves of Origanum syriacum in water and drying it using rotarvapor. MDA-MB-231 cell viability was tested by MTT assay as well as trypan blue exclusion in the presence or absence of increasing concentrations of OSE. Scratch assay as well as Boyden-chamber were used to determine effect of OSE on migratory capacity. Furthermore, the ability of MDA-MB-231 to adhere to fibronectin was investigated using adhesion assay. Phosphorylated ERK1/2 was measured using Western blotting. Results: OSE reduced proliferation of MDA-MB-231 cells in a concentration and time dependent manner. The optimum concentration was determined according to the significance of decrease in viability. Also, in the presence of OSE, there was a decrease in migration of cells. Furthermore, a dose-dependent inhibition of adhesion was seen in MDA when treated with OSE. Moreover, preliminary results indicate that OSE decreased ERK1/2 phosphorylation in MDA-MB-231 cells. Conclusion: O. syriacum may be considered a supplementary drug for patients with malignant breast cancer. Further studies should be conducted to elucidate the molecular mechanism of the anti-cancer property exerted by OSE.