Qatar Foundation Annual Research Conference Proceedings Volume 2014 Issue 1

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.