Qatar Foundation Annual Research Conference Proceedings Volume 2014 Issue 1

Background: Undetected repeated foot loading and temperature changes during walking in diabetic patients with peripheral neuropathy significantly increases risk of foot ulceration. Early detection of inflammation due to foot temperature and pressure changes from walking has shown to be a potentially effective strategy in prevention of foot ulcers. Therefore, a clinically feasible assessment of these changes is vital to measure pre-ulcerative inflammation and predict DFUs. The current study is aimed to validate effectiveness of an innovative fiber optics embedded smart textile for simultaneous measurement of plantar pressure and temperature. Methods: The study recruited 21 diabetic patients (Age: 57.8±7.9 years, BMI: 31.6±8.0 kg/m2, VPT: 26.8±15 volt, 68% diagnosed with peripheral neuropathy) at high risk for foot ulcers from Wound and Diabetic Foot Center at Hamad Medical Co. Doha. A series of sensors were juxtaposed on the length of an optical fiber integrated in a comfortable sock (Novinoor LLC, IL) for measuring temperature and pressure changes under anatomical regions of the heel, midfoot, 1st and 5th metatarsal heads, and big toe based on changes in wavelength of light. A thermal image was acquired after five-minute temperature acclimation before walking for validating temperature changes. Participants walked 200 steps to induce thermal stress and follow up thermal image was acquired. Participants also wore F-scan insoles (TekScan®Inc) during walk for pressure validation. Results: All the recruited patients perceived the device as comfortable. A significant correlation was observed between both pressure (r=0.67, p<0.05) and temperature (r=0.55, p<0.05) measurements between the sock and the reference systems under different anatomical regions of interest. Conclusion: This study demonstrates the proof of concept for an innovative smart textile in simultaneous assessment of the key parameters associated with risk of foot ulcer in patients with diabetes. Given the correlation values, additional studies will larger sample size is required to further validate and address whether the technology can predict and better manage diabetic foot ulcers. Funding source: NPRP 4-1026-3-277

Obesity is a growing health concern in Qatar because of the increased risk for type 2 diabetes, hypertension and other vascular disorders. One of the early adverse vascular events in obese individuals is an abnormal endothelial function which might alter the mechanism of vasodilatation. In particular, it is thought that loss of nitric oxide (NO)-mediated vasodilatation might be compensated by dilatation mediated by endothelium-derived hyperpolarizing factor (EDHF) in these individuals. It is however not clear whether such a switich in the contributions of these mediators would depend on the degree of loss of vascular endothelial function. Given that micro vessels in subcutaneous (SC) and omental (OM) fat depots of obese individuals might suffer different degrees of endothelial dysfunction [1], this study investigated whether the roles of NO and EDHF as mediators of vasodilatation of these vessels are altered in morbidly obese Qataris. Small arteries were isolated from SC and OM adipose tissues collected from consented morbidly obese Qatari patients (n=18) undergoing bariatric surgery at Hamad general hospital Doha. The vessels were studied by wire myography. Relaxation curves were generated for actetylcholine (Ach, the classical endothelium-dependent relaxant) in the absence or presence of Nω-Nitro-L-arginine methyl ester (L-NAME, 100 µM, inhibitor of nitric oxide synthase), apamin (0.5 µM) + charybdotoxin (0.1 µM) -combined blocker of EDHF, or BaCl2 (30 µM, blocker of inward rectifier potassium Kir channel ) on initial tone built with noradrenaline (1-5 µM). Curves were also generated for the NO donor and endothelium-independent relaxant, sodium nitroprusside (SNP). The patients had body mass index (BMI) of 46±2 Kg.m-2 and fasting plasma insulin of 17±2 mU/L [Mean±SEM]). Vessels from the SC depot were generally more responsive to Ach compared to OM vessels. Ach curves for both SC and OM vessels were significantly shifted to the right by L-NAME (n=5-7, p<0.05) and more so by apamine+charybdotoxin (n=6-7, p<0.01). The Emax for SC vessels dropped by ̴ 37 % in the presence of L-NAME and by ̴ 64 % in the presence of apamin+charybdptoxin. For OM vessels, the the reductions were ̴ 27 and ̴ 40 % respectively. BaCl2 caused a rightward shift in Ach response in both vessel types (n=4-5, p<0.05) although significant reduction in Emax was only recorded in OM vessels (p<0.05). When Ach and SNP responses were compared for each vessel type, only the OM vessels showed significant endothelial dysfunction (p˂0.01). The results show that eventhough NO still plays a significant part in the endothelium-dependent dilator mechanisms of these vessels from morbidly obese Qataris, EDHF appears to play a greater role, particularly, in the SC vessels. The results also show that the increased role for EDHF is not due to loss of endothelial function since the substantially greater role in SC vessels was observed inspite of the relatively better endothelial function recorded in them compared with OM vessels. Furthermore, Kir channel involvement appears to be depot specific and largely in the OM vessels. 1. Farb MG et al., 2012 Arterioscler Thromb Vasc Biol. 32(2):467-73

Background & Objectives: Epilepsy is a neurological disorder that is associated with repeated episodes of seizures. In epilepsy, the normal pattern of neural activity is disturbed, causing the patient to experience various symptoms ranging from staring blanking for a few seconds to long periods of vigorous convulsions and unconsciousness. In many patients, the injuries they endure are a direct result of the confusion, loss of muscle control, and unconsciousness caused by the seizure. These injuries include fractures, head injuries, and burns. In an attempt to mitigate such risks, extensive research has been dedicated to developing a device that can detect or predict the onset of seizure episodes. The clinical behavior of an epileptic seizure is preceded and then accompanied by electroencephalographic alterations. As a result, electroencephalography (EEG) is the most common tool to measure these alterations. EEG measures the voltage fluctuations resulting from ionic current flows within the neurons of the brain. Research has demonstrated that epileptic seizures are caused by disturbances in the electrical activity between neurons in the brain. In the healthy brain, neurons fire in an asynchronous manner, relaying messages from one neural network to the other. However, in the epileptic brain, the complex interactions between neuronal networks are characterized by the evolution of synchronization between them. Excessive neuronal synchronization leads to a hyper-synchronous state that triggers the onset of a seizure. Methods: There are many ways to measure the synchrony between two or more continuous time series of brain activity, ranging from linear approaches such as the cross correlation and the spectral coherence function as well as nonlinear measures like mutual information, transfer entropy, Granger causality, or nonlinear interdependence. These measures yield low values for independent time series and high values for correlated time series. In this research, we present a novel patient-specific epileptic seizure onset detector using scalp EEG. The proposed detector employs a novel neural synchronization measure to compute the level of EEG channel synchronization at a particular time instant. Training a support vector machine, in the classification stage of the detector, on the calculated level of neural synchronization from a patient's pre-ictal, ictal, and post-ictal EEG, the detector is able to identify the electrographic onset of an imminent seizure. Results: The proposed detector successfully identifies EEG epochs that are highly synchronized as seizure epochs, while pre-ictal and post-ictal epochs are shown to have a significantly lower level of synchrony. The performance evaluation results of the proposed detector are closely aligned with the electrographic seizure onset time that an expert has visually determined. Conclusion: The quality of life of epileptic patients that suffer from intractable seizures can be enhanced by equipping them with a device that can alert them ahead of time of an imminent seizure. The dramatic increase of synchronization between neuron firing during a seizure is clearly detected using the novel synchronization measure that we propose. The proposed seizure onset detector clearly identifies times of increased neural synchrony and sends an alarm at the earliest abnormal electrographic changes of a patient.

Oxygen is essential for the survival of all aerobic organisms but it is also, paradoxically, a toxic, mutagenic gas. Molecular oxygen is highly reactive and its partial reduction generates reactive oxygen species (ROS). Under normal physiological conditions, ROS are produced in low amounts as a result of active aerobic metabolism and have an important role as signaling molecules. At higher concentrations, ROS are cytotoxic and damage macromolecules. Proteins are major targets of ROS, resulting in a change in the structure and/or function of the oxidized proteins. Dietary antioxidants have been shown to modulate the extent of protein oxidation in plasma. However, the significance of dietary antioxidants in preventing vital oxidative changes of proteins still remains unknown.Therefore, in the present work we have evaluated the antioxidant significance of cucumber and zucchini extracts in the prevention of oxidative modification of proteins in human keratinocytes. Vegetables were collected from the farm in Qatar and their juice was extracted by a heavy-duty juicer, with a stone mill-like screw, using a low speed of 80 RPMs resulting in minimal heat build up and oxidation of samples. Copper sulphate, a well known inducer of lipid peroxidation, was used to alter redox homeostasis in human keratinocytes. Cells were treated with copper sulphate (LD22.5) and vegetable extracts were used to prevent oxidative damage of proteins. After 24 hours, the ability of extracts to inhibit copper induced cell death was evaluated by MTT assay and the proteins were isolated for redox proteomics analysis. Proteins were then separated by 1-D gel electrophoresis, digested with trypsin and peptides were detected by Orbitrap LC-MS/MS. Afterwards, the MS/MS data were searched using Proteome Discoverer 1.4 against Homo sapiens database incorporating the comprehensive workflow that included several dynamic modifications of peptides, like N-terminal carboxymethyl, C-terminal oxidation and terminal independant oxidation and trioxidation of peptides. The results obtained showed that cucumber and zucchini extracts were able to inhibit copper induced cell death, LD0 and LD12 respectively. Furthermore, the results obtained revealed several peptide modifications that were inhibited by the vegetable extracts and associated with inhibition of copper induced cell death. Finally, future identification and quantification of redox-based changes within the proteome both in redox signaling and under oxidative stress conditions and their inhibition by dietary antioxidants represents new and exciting field that will have a significant impact on a number of pathophysiological conditions and aging

Although prostate cancer can be clinically managed in its early phases, the inability to control the more aggressive late-stage disease has prompted the search for novel therapies. We hypothesize that strategies targeting polyamine homeostasis may be effective against prostate cancer. The prostate has the highest level of polyamine biosynthesis of any tissue, and it is the only tissue in which polyamines are purposely synthesized for export. In particular, large amounts of polyamines are excreted by the prostate into semen. Thus, we reasoned that polyamine homeostasis may be altered in the prostate relative to other tissues and that tumors derived from this gland may exhibit atypical regulatory responses in order to synthesize large amounts of polyamines in this tissue. Intracellular levels of polyamines are homeostatically maintained by effector systems controlling the biosynthesis, uptake and export of these molecules. By acetylating polyamines, the enzyme spermidine/spermine N1-acetyltransferase (SSAT) controls polyamine inactivation and export. Specifically, SSAT catalyzes the transfer of acetyl groups from acetyl-coenzyme A (acetyl-CoA) onto intracellular polyamines and thereby marks them for export and/or catabolism, which reduces their positive charge, blocks biological activity, promotes degradation, and facilitates excretion. We showed that in prostate tumors, overexpression of SSAT leads to futile metabolic cycling (polyamine flux). Furthermore, our and others studies have shown that overexpression of SSAT in transgenic mice (SSAT-tg) increased energy expenditure, which produced a polyamine-based cycle, reduced the levels of acetyl-CoA, increased glycolytic enzyme expression and decreased lactate production. These proof of principle studies provided evidence for linking the polyamine flux to the Warburg effect. Additional studies in prostate tumor cell lines have shown that the polyamine flux increases the glucose uptake, thereby creating the possibility of improved positron emission tomography (PET) with 18-fluorodeoxyglucose (FDG) because this imaging is not currently possible in prostate cancers. To further study the effect of polyamine flux on tumor growth in vivo, our transgenic mice were crossed with TRAMP mice (TRansgenic Adenocarcinoma of Mouse Prostate) to produce a bigenic mouse line (TRAMP/SSAT). These mice showed an increased in polyamine flux and importantly a reduction in prostate tumor burden. This novel approach for treating cancer by targeting a key regulator of cancer metabolism may provide a better, more efficient treatment for prostate cancer and improved PET imaging.

Insulin resistance and beta cell dysfunction are core defects in type 2 diabetes (T2DM). Impaired fasting glucose (IFG) and impaired glucose tolerance (IGT) are intermediate states in the transition in glucose tolerance from normal to T2DM, both of which are associated with increased conversion rate to T2DM. Understanding the metabolic abnormalities that lead to the development of IFG and IGT will help develop strategies to prevent the development of T2DM. in the present study, we have examined the metabolic abnormalities responsible for the development of IFG and IGT in Arab individuals. 43 subjects with NGT (n=17), isolated IFG (N=17) and isolated IGT (n=9) received 75-gram OGTT and plasma glucose, insulin and C-peptide concentrations were measured at baseline and every 15 minutes for 3 hours after the glucose drink. Insulin secretion was measured with the incremental area under the plasma insulin and C-peptide concentration curves and insulin sensitivity was measured with the Matsuda Index. Beta cell function was measured with the Disposition Index as the product of insulin secretion and insulin sensitivity indices. IGT subjects manifested severe insulin resistance compared to IFG and NGT subjects. The Matsuda Index was 3.7±0.4, 7.2±1.0 and 8.4±1.1, respectively (p<0.01 for IGT versus both IFG and NGT). Beta cell function measured with the Disposition Index from 0-180 minutes was markedly reduced in IGT subjects compared to IFG and NGT individuals, 6.5±1.2, 34.0±5.6 and 32.9±4.9, respectively, (p<0.05 for IGT versus IFG and NGT. However, insulin secretion during the first 30 minutes during the OGTT was reduced in both IFG and IGT compared to NGT, 68.1±8.1, 52.6±7.4, and 100.7±10.9, respectively, (p<0.01 for both IFG and IGT. Collectively, these results indicate that similar to other ethnic groups, distinct metabolic defect characterize IFG and IGT in Arabic individuals Acknowledgment: We would like to thank Sanaa Mansy and Evette Rofaeil for their excellent care of the study participants. Supported by Qatar Foundation grant NPRP 4-247-3-076 to MAG and MZ

Qatar experience in Standard Breast Cancer Screening Authers: Dr Salha Bujassoum ,Dr Hekmet Bugrein , Dr Reena ALaSSAM , Dr Mufid Elmistiri Abstract Qatar has one of the highest age-adjusted breast cancer incidences in the Arab world. Although this is much lower than the incidence in the West. Breast cancer incidence in Qatar was 45 per 100,000 in 2003-2007.These higher incidence rates in Qatar are mainly due to the growing population. The prevalent age group, between both Qatari and non-Qatari patients, was 40-50 years old. This suggests that the age-specific incidence of breast cancer in Qatari women is shifting more to a pattern usually not seen in Western nations where median age at diagnosis is 61 years of age, moreover the diagnosis is often at advanced stages of breast cancer. These factors, together with reconfirmed evidence of mortality benefit from breast cancer screening trials, led to establishment the first hospital based mammographic breast cancer screening program in Qatar. It uses a distributed model of mammography service. The program accepts eligible asymptomatic women at age between 45 - 69 years and was launched in 2008. It adopted international standards of breast screening practice and breast cancer detection. Methods A retrospective study was conducted to describe the breast cancer screening program in Qatar and to overview the detection rates, positive predictive values and sensitivity and specificity of mammogram. Results Total number of screened women was 4264 with an increasing participation, year by year. Out of these, Qatari patient's accounts for 1145, and non Qatari for 3119. The age group of breast cancer detected cases from screening program (43-51). Total breast biopsies were 82 core, of which 45 were positive of breast carcinomas, (37) invasive ductal carcinoma, (8) noninvasive ductal carcinoma. The Invasive cancer detection rate was 8.2 % The positive predictive value (PPV) was 46%. Sensitivity value has improved from 51% in 2008 to 70% in 2012 as well as specificity value that has increased from 77% in 2008 to 83% in 2012. Conclusion Public acceptance of the breast cancer screening program in Qatar gradually increased and detection rates were acceptable in this part of the world. We have a unique population of multinationals that merits tailored screening tools.

The eye is the clearest and most accessible organ to observe anatomical and physiological characteristics of microvessels, the smallest blood vessels of the circulatory system. The microcirculatory bed of the retina shares similar anatomical and physiological characteristics with the cerebral and coronary circulations. Therefore, subtle changes in the retinal blood vessels can mirror early cardio- and cerebrovascular events long before their clinical manifestation (e.g. hypertension). Retinal images contain information to develop quantitative diagnostic algorithms for morphology, physiology, and pathology. Analysis of retinal images allows quantification of many vascular features: the Central Retinal Arteriolar and Venular Equivalents (CRAE & CRVE), morphological features (branching, tortuosity and geometric pattern of the vascular bed), but also small changes in the diameter of microvessels in response to disease or external factors. Particularly CREA and CRVE have been associated with hypertension, coronary heart disease, stroke, and type II diabetes as well as with acute responses to environmental (e.g. air pollution exposure) and lifestyle (e.g. smoking and physical inactivity) risk factors. We demonstrated in a longitudinal panel study with 52 healthy individuals aged 22 to 62 years that short-term changes in particulate air pollution (PM10-concentrations ranging from 9.16 µg/m³ to 126.58 µg/m³) caused a significant narrowing of the CRAE by 0.69 µm (95% CI= -1.13 µm to -0.26 µg; p = 0.0032) on the day following peak exposure. This narrowing was equivalent to 1.5 years of aging. The associations were independent of individual characteristics such as gender, age and BMI. The results indicate that microcirculatory blood vessels can be affected by short-term changes in air pollution, and may contribute to the development or aggravation of cardiovascular disease, perhaps even induce events. To allow for a more automated analysis of retinal images, we developed stand-alone software which results in a faster, objective, and more precise grading of large numbers of images. VITO embeds this tool in a data portal, which includes batch processing and quality control. We are exploring if handheld retinal cameras can replace heavier and more expensive tabletop cameras. If these instruments are interchangeable, then we can work towards a truly mobile health solution. The unobtrusive nature of retinal imaging has high potential for facilitating long-term, recurrent monitoring of the population in epidemiological settings, in line with the P4-Medicine paradigm (Preventive, Personalized, Participatory & Predictive). Also for specific vulnerable subpopulations such as elderly, pregnant women, and school children, the non-invasiveness and ease of routine allows for wide deployment in large-scale, even population-wide longitudinal studies. Retinal imaging can also contribute to evidence-based population health programs. It addresses several pressing health challenges in Qatar and Belgium (cardiovascular diseases, obesity and diabetes); identifies risk factors and their effect in vulnerable groups (e.g. the potential adverse health effects of exposure to indoor and/or outdoor air quality) and provides a non-invasive and personalized method to follow-up the effect of disease prevention and healthy behavior campaigns. VITO is actively seeking for partners and projects in this research area.

A gradual shift from GHz to THz range in the past few years has celebrated the invention of many new technologies in the field of communications, computers, electronics and medicine. Nano-networks are the new era technologies defined as electronic components and their interconnection within a single chip on a nano-scale [1]. The concept of Network on Chip (NoC) is exploited on a scale ranging from micrometers to nanometers. This term is also referred to as the network-like interconnection of nano-materials such as carbon nanotubes arrays [2,3]. The research done in the past has broadly explained the same concept of nano-networking with different terminologies and slightly varying definitions depending on the mechanisms of networking and communication. In this study, the main focus is on propagation investigation at THz frequencies in human tissues, such as blood, fat, and skin, which is conducted with the aim to fix the early paradigms for a pioneering investigation in this field. The path loss values obtained from numerical simulation have been compared to an analytical model in order to verify the feasibility of the numerical analysis which can be subsequently extended in order to take into account more realistic parameters such as the antenna, or inhomogeneity of the materials. The study also explores the methods and techniques to characterize biological medium necessary for Nano-network communication. Time Domain Spectroscopy has immensely contributed in the field of medical imaging, analysis of protein and other biomolecules, finger print analysis of chemicals and food quality control [4]. The aim is to utilise this technique to study the dielectric properties of various biological samples. Subsequent sections of the chapter will discuss various biological entities such as skin & its layers, blood, hydration shell and sweat ducts. As one begins to understand the human body anatomy from wireless communication perspective, one can realizes its overall complexity and random nature. The main objective of this work is the potentiality of nano-scale EM communication exploiting the THz regime of the electromagnetic spectrum. The nano-scale EM communication becomes an obvious choice due to the fact that molecular information for a human body is indeed sensitive in the THz regime. Additionally, this part of the spectrum is much safer to interact at a cellular scale that its counterparts in the microwave or gigahertz range. The proposed in-vivo body nano-networks ensures their stability without disturbing the harmony of in-built molecular structure of the body. Moreover, in most of the cases medicine fails to understand the root cause of the problem but once a monitoring network is established, one can extract various unknowns and treat them effectively.

Rationale: Type 1 diabetes (T1D) is characterised by autoimmune destruction of pancreatic β-cells. Enterovirus (EV) infections have been frequently linked to T1D, but a causal relationship has not been well-established. MicroRNAs (MiRNAs) function as post-transcriptional regulators of gene expression. However, their role in virus-induced β-cell death has not been investigated. We hypothesize that EV infection of β-cells alters both cellular and circulating miRNAs abundance, thereby inducing specific gene expression patterns which result in β-cell death and T1D pathogenesis. Objective: To investigate the association between EV infection and miRNA dysregulation in human islets infected with EVs, and to develop circulating miRNA signatures to detect and distinguish EV-infection in T1D patients. Methods and Results: (a) Primary human islets were infected with clinical EV strains, including Coxsackievirus (CV)A9, CVB2, CVB5, isolated from stool specimens of children at seroconversion to islet autoimmunity. Cells/cell culture supernatant were collected at D0-to-D14 post-viral infection to measure viral replication, cytopathic effect (CPE) and cellular miRNAs; (b) Plasma samples from children with T1D and/or EV infection were tested for circulating miRNAs among different between-group comparisons (EVpositive/T1D; EVnegative/control; EVpositive/control and EVnegative/T1D). RNA was extracted using (a) miRVana kit and (b) Trizol-based method. All samples were tested using global miRNA profiling (TaqMan OpenArray, QuantStudio12K Flex). In addition, 11 individual miRNA were tested (Quantitative realtime-PCR, TaqMan). 'miRBase', 'TargetScan' and 'miRWalk' miRNA databases were used to identify validated and predicted miRNA gene targets. DAVID Functional Annotation tool was used for gene-enrichment analysis of the putative target genes. All EV clinical isolates replicated and demonstrated CPE in human islets. Differential expression of individual miRNAs was demonstrated in the infected islets vs no-virus control (NVC) islets. The expression of miRNA-627, miRNA-302a, miRNA-190b, miRNA-497, miRNA-888, miRNA-124* and miRNA-340* were significantly higher in CVB2-infected islets vs NVC (> 10 fold difference). In addition, subsets of circulating miRNAs were found to be differentially expressed between the study groups. Of 434 miRNAs detected in at least one plasma sample, 11 miRNAs were significantly different between the 4 groups and between T1D vs controls (miR-539, miR-532, miR-886-5p, miR-125a-5p, miR-340, miR-574-3p, miR-28-3p, miR-150, miR-339-3p, miR-151-3p) (p-value <0.05). MiRs miR-376a, miR-629, miR-140, miR-345, miR-146b, miR-222, miR-146a were also significantly different between T1D vs Controls. miR-139-5p was significantly altered across the 4 groups, and between EV-positive vs EV-negative groups alongside miR-136*, miR-744*, miR-15a*, miR-148a, miR-379, miR-410, miR-223*, miR-93*, miR-342-3p, miR-885-5p, miR-520c-3p. Moreover, 13 miRNAs were detected in all samples and groups in the initial study, however only miR-151-3p was significantly differentially expressed. Gene-enrichment analysis revealed apoptotic and inflammatory cytokine signaling to be major biological pathways that are regulated by the miRNAs detected. Conclusions: Viral infection of human islets leads to dysregulation of multiple miRNAs that may have a critical role in immune modulation, thereby contributing to β-cell death in T1D. Such EV-associated T1D may also be detected in altered circulating miRNAs. Measuring circulating and cell-associated miRNAs will greatly further our understanding of virus-induced T1D, which could have broad applicability as a disease biomarker.

Background Bacterial superantigens (SAGs) are potent T cell stimulatory molecules and comprise a large family of disease-associated proteins. Superantigens bind to APCs on the outside of the MHC class II molecule and to T cells via the external face of the T cell receptor (TCR). This enables them to activate up to 20% of resting T cells, whilst conventional antigen presentation results in the activation of 0.001- 0.0001% of the T cell population. These biological properties of superantigens make them very attractive for use in immunotherapy. Objectives 1.Codon optimization, for maximum expression in E. coli and synthesis of four superantigens genes 2.Cloning, Overexpression, purification and molecular characterization of four superantigens Methods The four superantigens codon-optimized genes were produced by Geneart GmbH Company (Regensburg, Germany). Additionally, we introduced several new restriction enzyme sites into the gene, e.g. NdeI and HindIII sites were placed at the 5' and 3' ends of the gene, respectively to facilitate the cloning. Each gene was excised from pGA/sAg by digestion with NdeI and HindIII and inserted into similarly digested pET28a DNA using T4 DNA ligase. The newly formed construct of each gene was transformed into competent cells of E. coli BL21 (DE3) RIL. The expression of each superantigen was done by induction using IPTG. The induced cells were harvested by centrifugation and the total proteins were produced by sonication. The expression analysis was performed using SDS-PAGE. Novel SAGs (TSST, SPE, SEA and SEB) were tested in vitro for their superantigenicity and anticancer activity using peripheral blood mononuclear cells (PBMC) isolated from healthy volunteers and prepared by Ficoll-Paque gradient centrifugation.Superantigenicity was assessed by monitoring the SAG-induced release of PBMCs cytokines. PBMC were treated in the presence or absence of SAGs for 24 h and the release of IL-1β, IL-8, IL-10, TNFα and IFNγ measured by Luminex assay. Furthermore, 3 different colon cancer cell lines were treated with SAGs in the presence or absence of PBMC for 48 h. Inhibition of cancer cell proliferation was measured by MTT assay. Results In this study, four synthetic genes coding for four different superantigens, SEA, SEB, TSST-1 and SPEA were codon-optimised and synthesized for maximum expression in E. coli using the vector pET28a. Our work showed that in all cases the recombinant superantigens were expressed to 40% of the total host protein. We managed to optimize the condition to obtain 20% of the recombinant superantigens in a soluble form. The purification of the soluble His-tagged recombinant superantigens have been achieved in a single step by Ni2+ charged column chromatography. Superantigenicity assay by measuring the released interferon gama showed that the four recombinant superantigens are active with superantigenicity functions. Conclusion The successful cloning and expression of four codon optimised synthetic superantigens genes will provide the recombinant protein which will be modified for production of safer superantigen for cancer immunotherapy.

Background: Healthcare in Qatar is undergoing a period of major reform, driven by a strong economy and vision for a world-class healthcare system. One area identified as a potential contributor to developing a world-class healthcare system is interprofessional education (IPE), with the goal of facilitating collaborative practice among healthcare workers. Several key steps have been taken towards developing IPE in Qatar, such as the formation of the Qatar Interprofessional Health Council (QIHC), the development of an IPE program for undergraduate healthcare students, the development of a set of shared core competencies, the receipt of substantial buy-in from leaders across the healthcare system, and recent approval of funding to develop a post-licensure healthcare IPE program. In order to improve IPE in Qatar, it is important to better understand the facilitators and barriers to interprofessional collaboration in Qatar. This study seeks to do so by qualitatively exploring facilitators and barriers to interprofessional collaboration in Qatar from the perspective of healthcare professionals. This research can assist in improving healthcare in Qatar by increasing understanding about how healthcare workers give meaning to interprofessional education and collaboration. Objectives: The purpose of this paper-presentation is to report on finding from a qualitative study that explored different facilitators and barriers of interprofessional practice in Qatar. Method: 10 healthcare professionals who work in Qatar were interviewed using semi-structured, open-ended interviews organized by the theoretical approach of phenomenology. The interviews focused on exploring barriers, facilitators, and what is working well in terms of interprofessional practice for healthcare professionals in Qatar. Findings and Implications: Different factors associated with interprofessional collaborations will be discussed. In doing so, this research adds to the literature on IPE by shedding light on interprofessional collaboration and education in the Middle East. Furthermore, this study identifies barriers for healthcare workers to work collaboratively in healthcare settings in Qatar. Addressing such barriers, and building on what is working well, will facilitate Qatar in reaching one of the Qatar National Vision 2030 goals of improving Qatar's health and wellness.

Background & Objectives The state of Qatar has witnessed significant lifestyle changes due to rapid urbanization, the introduction of labour-saving devices and the availability of high-caloric density food. This has impacted on the daily lifestyle and health habits of young adults leading to significant increases in non-communicable diseases (WHO, 2014). This study explored the risk factors associated with such diseases amongst young adults in Qatar. Methods A representative sample of 732 males and females (aged 18-25 years) from Qatar University took part in this cross-sectional, mixed-method design study. Physical Activity (PA) and dietary habits were assessed using a validated questionnaire. Total energy expenditure per week was calculated based on the metabolic equivalent values of each activity reported by the participant (Al-Nakeeb et al., 2012). Body Mass Index (BMI) was calculated according to the International Obesity Task Force criteria and using the age and gender-specific BMI classification established by Cole et al. (2000). Results The percentage of overweight/obesity in males and females was 39.5% and 38.5% respectively. It was evident that there was a significant increase in the percentage of students classified as overweight/obese from year 1 to year 4. Meanwhile, there was a decline in the level of PA and an increase in sedentary time during that period. Whilst health was reported to be the main reason for participation in PA/sport, lack of available time was singled out as the main barrier to engagement in an active lifestyle. Ironically, students reported more than 4 hours of TV/DVD viewing and internet use per day. Conclusions The adoption of healthier lifestyles amongst the Qatari population, including an increase in PA and a reduction in overweight/obesity are major objectives cited in Qatar Vision (2030). This study has revealed a high prevalence of overweight/obesity amongst male and female university students with regressive trends in their lifestyle and health habits. The findings reveal a worrying picture of young people's lifestyle that ought to be a cause for concern for policy makers and health professionals. Undoubtedly, there is an urgent need to seriously consider putting in place intervention strategies concerning behaviour modification and the built environment in order to reverse these trends. Such strategies could have major implications on the health and well-being of young people at this critical age developmentally and on the future welfare of the wider community in the long run. References *WHO (2014) World Health Statistics - 2014. WHO Press: Switzerland. *Al-Nakeeb, Y., Lyons, M., Collins, P., Al-Nuaim, A., Al-Hazzaa, H., Duncan, M. and Nevill, A. (2012) Obesity, physical activity and sedentary behavior amongst British and Saudi youth: A cross-cultural study. Intl J Environ Res Public Health, 9, 1409-1506. *Cole T.J. et al. (2000) Establishing a standard definition for child overweight and obesity worldwide: International survey. Br. Med. J., 320 (7244): 1240-1243. *General Secretariat for Development Planning - Qatar National Development Strategy 2011-2016, Towards Qatar National Vision 2030. Qatar: Gulf Publishing and Printing Company, Doha.

Protein misfolding and amyloid formation is an underlying pathological hallmark in a number of prevalent diseases of protein aggregation, including Parkinson's disease, Alzheimer's disease and Type 2 diabetes (T2D). The expansion in the prevalence of T2D, including in Qatar, where a high percentage of the population is affected by diabetes, poses considerable risks to individuals, the healthcare system and the economy. Epidemiological studies reveal that up to 95% of all patients with T2D are shown to have pancreatic amyloid deposits, as detected in post-mortem studies. Most importantly, the severity of the disease appears to correlate with the degree of the deposition of IAPP aggregates. The misfolding of IAPP followed by the aggregation has been shown to be highly cytotoxic and to play a key role in the death of β-cells in T2D. Thus, the modulation of the aggregation process, promoting the proper folding of IAPP, may be considered as an attractive avenue for a therapeutic intervention. Indeed, molecular chaperones have been shown to suppress the misfolding and to prevent amyloid formation. The mechanism of how Hsc70 inhibits hIAPP fibril formation is complex and is not yet fully understood. It remains unclear how specific domains of Hsc70 function independently or in cohort to produce the observed inhibition of fibril formation of hIAPP. To address these questions, we used the power of in vitro methods to dissect out the relative contribution of the different Hsc70 structural domains, by investigating the effect of a series of deletion mutants of Hsc70 on hIAPP fibril formation. We also investigated in further detail the mode and mechanism of interaction between Hsc70 and hIAPP. The results indicate that Hsc70 can bind to hIAPP and delay fibril formation even in the absence of the ATPase domain, but interaction of hIAPP with the substrate- binding domain is strongly influenced by the C-terminal lid region. Determining the molecular interplay between hIAPP and molecular chaperone Hsc70 will provide the basis for designing modulators of IAPP aggregation.

Background: Sickle Cell Anaemia (SCA) is a genetically-inherited blood disorder caused by the occurrence of a point mutation in the bases coding for the sixth amino-acid of the β-chain of haemoglobin. The presence of Fetal Haemoglobin (HbF) in blood is known to show a number of beneficial effects in improving the conditions of SCA. Hence, clinical symptoms of SCA arise only after HbF levels drop. HbF is synthesized by the HBG1 (gamma-globin) gene. From birth there is a gradual shift from fetal to adult haemoglobin triggered by the downregulation of the HBG1 gene and the upregulation of the HBB gene. Rationale: Hydroyxurea has been widely evaluated in the treatment of SCA for its ability to induce synthesis of HbF by reactivating the expression of the HBG1 gene. It functions by repressing the activity of various transcription factors involved in the downregulation of the HBG1 gene during transition to adult haemoglobin. However, hydroxyurea has proven to be potentially toxic in a number of cases and may also carry a possibility of long term carcinogenicity. Therefore it is administered under restriction and may not be used in the treatment of pregnant women and children. In this study, an attempt is made to identify and characterise the altered gene expression pattern caused by hydroxyurea treatment through Differential Display Reverse Transcriptase PCR (DDRT-PCR) and Real Time PCR (Q-PCR). Further, the phenomenon depicting the repression of the transcription factors leading to the reactivation of HBG1 gene will be reproduced using techniques of RNA interference (RNAi) and antisense. Objective: To reproduce the altered genetic conditions produced by Hydroxyurea treatment for the in-vitro induction of HbF using molecular genetic techniques. Methods: Blood is obtained from SCA patients at diagnosis and after treatment with hydroxyurea. Erythroid Progenitor cells are purified from the collected blood using the EasySep™ Human progenitor cell enrichment kit. Cells are then cultured in a suitable medium and maintained for further analysis. Treatment with hydroxyurea is performed at varying doses at different intervals of time. The evaluation of gene expression is carried out through the generation of c-DNA through a reverse transcriptase PCR cycle followed by Q-PCR for identification and quantification. Results & Conclusion: Buffy coats containing mononuclear cells were purified from peripheral blood obtained from SCA patients through density-gradient centrifugation. Further, erythroid progenitor cells were successfully isolated and maintained in culture for variable periods of time. Currently, mRNA expression analysis is ongoing through the use of reverse transcriptase PCR protocols. If proven successful, this technique holds a promising future scope; as molecular genetic techniques could be used to reproduce the effects of hydroxyurea without the actual administration of the drug. Thus, the toxicity issues of the drug are avoided while also making it available to all classes of patients including pregnant women and children.

Background: Strokes account for over 8% of deaths in men and 12% of deaths in women in the UK and the total cost of Stroke to the National Health Service within the UK is estimated to be over £7 billion per year. Carotid artery disease can cause stroke, transient ischaemic attack (TIA) and amaurosis fugax. Surgical intervention to remove the carotid plaque is likely to be helpful in symptomatic patients with > 50% stenosis of the carotid artery. The aims of this study were to (i) investigate referral pattern of carotid artery disease for duplex vascular ultrasound in a local hospital in UK; (ii) investigate whether confocal microscopy can be used to obtain 3D images of the micro-structure of atherosclerotic carotid plaques; (iii) construct a blood flow model with a view to investigate how forces resulting from fluid flow interact with structural stability of carotid atherosclerotic plaque Methods: A one-year audit for the total number of carotid artery vascular ultrasound scans referred to the Medical Physics Department by all specialists was carried out between (January- December). Those with > 50% stenosis (using St Mary's criteria) were identified, including those with symptoms who underwent carotid endarterectomy (CEA) . Carotid plaques were collected from routine carotid endarterectomy surgery and examined using flow-modelling, bright-field and Laser-Scanning Confocal-Microscopy (LSCM) to generate 3D image datasets and visualisations of surgically removed carotid plaques. Results: Over a one-year period, the total number of carotid artery referrals by all specialists for ultrasound scanning at the Medical Physics Department was 1353; 63.3% (n=856) of whom were referred from the TIA Clinics by stroke specialists. Out of this total referral, there were 139 patients (10.3%) with > 50% carotid artery stenosis, 57 of whom were symptomatic and underwent surgical intervention (CEA). 3D imaging of carotid plaques, using LSCM showed that most specimens were predominately composed of lipid material, comprising necrotic core of amorphous debris and cholesterol clefts, with varying degrees of fibrous tissue present in all plaques. Regions of actual fibrous cap disruption and some ulceration were also seen. Fraying of the fibrous cap was notable with fibrous cap erosion and exposure of underlying necrotic core to lumen. Evidence of carotid plaque vulnerability as demonstrated by reduced fibrous cap thickness and large lipid-necrotic core with evidence of cracking was also seen in the 3D visualization. A blood flow simulation model shows how blood velocity changes could occur associated with reduction in lumen diameter caused by the plaque. The degree of carotid artery stenosis measurements obtained from these flow models were consistent with, and comparable to the degree of stenosis measurement recorded on the pre-operative vascular ultrasound reports of patients from whom these plaques were taken. Conclusions: The visualization of the internal 3-dimensional microstructure and geometry of the plaques could help in: (i) investigating key features that affect plaque structural stability; (ii) comparing 3D microstructure of the plaque with clinical imaging assessment and blood flow investigations; and (iii) developing markers to identify patients requiring clinical intervention.

Background & Objectives: Integration of Clinical Decision Support Systems (CDSS) in mobile and smart environments helps to improve the quality of life of people with health problems. CDSS are used to derive clinical conclusions from patient data, in order to automate and help the process of diagnosing and treating the patient. One way that CDSS can be implemented is to formalize a clinical guideline (document detailing best practices for diagnosing and treating patients) and use it on a knowledge base containing the patient's data. An interesting perspective is to use CDSS in a smart home (SH) setting, where data for the remote CDSS can be obtained from SH services and mobile devices using ambient and wearable sensors. However, in order to maintain minimum quality of service for CDSS decision support, the CDSS decision process must be deployed locally as a SH service an on mobile devices. An ideal example domain for this integration scenario is the diagnosis of sleep apnea. Sleep Apnea has several symptoms that include recurrent awakening, loud snoring, choking episodes, non-restorative sleep and daytime sleepiness. Usually, an individual with sleep apnea is not aware of having difficulty breathing, and is often recognized by others witnessing the individual during sleep apnea episode or is suspected because of the observed symptoms. Sensors could detect such episodes automatically without the need for a human intervention, or recognitions of said symptoms. The objectives are to illustrate the feasibility of CDSS as SH service and on mobile devices by using the Sleep Apnea CDSS. Methods: The sleep apnea CDSS decision process uses Semantic Web tools and rule-based reasoning, in order to formalize the current Canadian guideline for the recognition of sleep apnea. A total of 9 rules were derived. A patient dataset comprises health factors related to sleep apnea, including clinically relevant personal information, clinical measures and observations, and symptoms specific to sleep apnea. To validate the decision process of a CDSS integrated with smart homes, we implemented the Sleep Apnea CDSS decision process on an Android smartphone (Samsung Galaxy SIII). For this validation, we assume that we have received data from the patient diary application, the SH services or local smartphone monitoring services. We generated datasets containing clinical data (measurements), whereby fact values were created based on ranges encompassing both clinically normal situations as well as abnormal situations. We have 7 dataset configurations (1 to 7 days of data), with 10 generated datasets by configuration (70 datasets). Results: The validation shows promising results for using CDSS on mobile phone: loading data and rules (140-425 ms), executing rules (45-82 ms) and memory usage for the reasoning process (174-350 KB). Conclusions: The results show that a guideline compliant CDSS system can be implemented on currently available mobile devices. While this validation was limited in scope, as we did not tackle the precise derivation of sensor data into the used clinical facts, it shows nonetheless that with such set of inferred clinical facts, interesting and clinically relevant problems can be tackled.

Back ground Antibody Directed Enzyme Prodrug Therapy (ADEPT) is a technique which has been used in cancer treatment. This therapy consists of two steps the aim of which is to convert a prodrug to a powerful cytotoxic drug only in the vicinity of the tumor. The technique requires a bacterial enzyme, glucarpidase (former name: carboxypeptidase G2, CPG2). The use of glucarpidase in ADEPT and in detoxification of the cytotoxic methotrexate, MTX is needed to be done in several cycles. This however, hampered by the induced human antibody response to the glucarpidase. The technique therefore will benefit from novel glucarpidase which might avoid the human immune system. Objectives 1.Collections and screening soil samples for novel glucarpidase producing bacteria 2.Characterisation of the isolated bacteria 3.Cloning, overexpression, purification and the molecular characterisation of the novel glucarpidase Methods Isolation of novel glucarpidase producer(s) Over one hundred soil samples were collected from different fields in Qatar where folate rich fruit and vegetables are routinely cultivated. Screening of these samples was performed using folate as the sole carbon and/or nitrogen source. Several molecular techniques were used to characterize the isolated strains. Cloning of the novel glucarpidase gene The gene was cloned by production of genomic library of the novel strain chromosomal DNA and PCR techniques. The gene was subcloned in the overexpression vector pET28a and transformed into E. coli (DE3). The novel gene was expressed by induction using IPTG. The recombinant protein was purified using Ni2+ column. The activity assay was carried out using methotrexate as substrate. Results We successfully isolated two new glucarpidase producing strains, Stenotrophomonas Sp. AA1, and Pseudomonas Sp. AA2. The CPG2 gene from Stenotrophomonas Sp. was cloned and expressed in E. coli using pET28a vector. The SDS analysis showed that the gene was expressed in the soluble form to about 30% of the total protein. The glucarpidase gene was cloned so as to place a polyhistidine tag on the N-terminus of the enzyme. A one-step purification protocol was usually sufficient to obtain essentially pure protein for detailed enzyme assay and characterization. Previous studies indicate that the native form of glucarpidase is Zn2+-dependent. Our data indicate that the recombinant enzyme showed no activity towards the substrate methotraxate in absence of Zn2+ ions. Full activity was restored to the zinc-free enzyme by the addition of Zn2+ to the assay mixture. Therefore, the formation of active enzyme was shown to be dependent on Zn2+. Conclusion We successfully isolated two new glucarpidase producers. We managed to clone express and purify the new glucarpidase gene from one strain. Molecular studies on the novel glucarpidase have been carried out. The novel glucarpidase will pave the way for further application in the cancer drug detoxification and Antibody Directed Enzyme Prodrug Therapy techniques.

Background hypertrophic Cardiomayopathy is an inherited heart muscle disease with considerable heterogeneity at genetic and phenotypic levels and poor correlation between genotype and phenotype. Next generation sequencing could help in addressing this problem. Subjects and Methods The present study involved 144 unrelated consecutive index HCM patients enrolled in the BA HCM National Program in Egypt subjected to large scale high throughput targeted next generation of coding and exon flanking regions of over 100 genes involved in inherited cardiac conditions (ICC) including genes with a known role in HCM (Illumina HiSeq) Results Putative pathogenic variants were detected in 67 samples (67/144, 46%), mostly found in sarcomeric genes in the order of: MYBPC3 (38.8%), followed by MYH7 (33%), MYL3 (9%), TNNT2 (4.5%), TNNI3 (in 2 samples). TNNC1, MYLK2 and TPM1, ACTN2, TCAP, PLN, ACTN2, MYH6 showed a potentially pathogenic in 1 patient, each. Validation was performed by Sanger sequencing. Twenty six variants were novel (26/144, 18%) and were not found in any of the control series and by in silico analysis were scored as potentially pathogenic, hence were considered as variants of unknown significance and should be tested for coseggregation. An approximate of 10% (14/144) of the samples revealed likely pathogenic variants in genes not commonly known to be linked to HCM. Complex heterozygosity involving sarcomeric and non sarcomeric genes particularly ion channel genes was observed in almost a quarter of the samples in the present cohort (35/144, 24%), and homozygousity was found in nine patients(9/67, 9%). This could provide an explanation to the heterogeneity of the phenotype observed amongst different members within the same family and among unrelated patients having the same sarcomeric mutation. The studied cohort involved six HCM phenocopies 3 had mutations in PTPN11 and two cases had novel possibly pathogenic variants in RAF1. One Fabry case with a reported pathogenic mutation in GLA gene. Conclusion High throughput NGS has enabled detection of possibly/likely pathogenic variants in over 60% of samples studied. The detection of homozygous patients and complex heterozygosity in almost a quarter of the HCM patients provides a possible explanation to the phenotype heterogeneity observed in HCM.

Background Obesity is an inflammatory disease associated with immune cell defects. The objective of this study is to characterize the adipose tissue macrophage (ATM) phenotype and function in human omental adipose tissue and peripheral blood in relation to obesity and its changes after gastric sleeve surgery for weight loss. Methods and subjects: Adipose tissue was obtained from morbid obese subjects with BMI > 40 kgm2 subjects undergoing the surgery in the Metabolic/Surgical department at Hammed Medical Hospital (HMC)-Qatar. Metabolic markers were measured in fasting serum and immune blood cells of the peripheral blood and ATMs were characterized by flow cytometry For immunophenotyping of T- cell populations of CD4+ cells, the circulating CD 45+ was counted using (BD LSRFortessa TM Cell Analyzer) and was utilized for WBCs subpopulation. CD4+ T-cell subpopulations were defined as naïve (CD45RA+andCD27+), central memory T cells (CD45RO+andCD27+), and effector/peripheral memory (CD45RO+and CD27-) and natural T- regulatory cell (CD4+CD25+ Fox3+). ATM was analyzed for CD11c and CD206. Results The mean and SD age of the study subjects were 31.67 (9.80) years, and females were more frequent 73.3%. Follow up of the study subjects after surgery after 3 month revealed that BMI was significantly reduced by 8%. Counting of the CD4+ T- cells subpopulations by flowcytometery on isolated PMBCs revealed that gastric sleeve significantly reduced the naïve cell number (CD45RA+andCD27+), central memory cells (CD45RO+and CD27+) and natural T- regulatory cell (CD4+CD25+ Fox3+), but not effector memory cells (CD45RO+and CD27-) among our study subjects. ATM shows marked distribution of CD11c+ CD206+. Based on the HOMA insulin resistance calculation, 67.0% were insulin resistance (IR) and had higher mean and SD of glucose in mmol/L (6.74 ±0.93) than non-IR subjects (5.58±1.24] with p value=0.106 but had significantly higher mean and SD of insulin level µU/ml in IR subjects (24.75±4.08) than non-IR subjects (13.75±3.77] with p value=0.006. A significant reduction of circuiting Il-6 and MCP-1 (markers of M1 polarization) was observed after the intervention by 33% and 13.0 % (p=0.021 and p=0019), respectively, while Il-10 a marker of M2 polarization significantly increased by 12.0% after surgery with p =0.050. Conclusion Weight loss intervention to morbid obese subjects by gastric sleeve with energy restriction was accompanied by a significant change in immunophenotyping polarization from M1 to M2 with a shift from pro-inflammatory to anti-inflammatory pattern.

Personalized medicine uses information about an individual's genes, proteins, environment, and phenotype data to prevent, diagnose, and treat diseases. In addition, the innovative bio-markers discovery as the key of personalized medicine across multiple tumor types has unlocked new information about cancer biology by providing critical insights to biological, pathogenic and pharmacologic responses to treatment. In this decade, the finalization of the human genome project, when a complete sequence was published for the first time, created the potential to identify a large set of single nucleotide polymorphisms (SNPs) across the entire genome. Consequently, this has opened the door to possibilities for great improvements in diagnosis and therapeutics. In addition, the availability of massive amounts of Genomic Wide Association Study (GWAS) data has necessitated the development of new data mining and machine learning methods for quality control, imputation and analysis issues including multiple testing, predictive modeling for chronic diseases, and to discover variants that could lead to a particular trait/disease. Currently, personalized medicine faces multiple issues when trying to predict complex diseases such as cardiovascular, cancer, and asthma…etc. Yet, disease prediction still based on SNPs and few environmental factors, while complex diseases are usually affected by gene-to-gene interactions and many environmental factors which have great impact and significance on the predicted outcomes. Therefore, the current challenge is to develop a unique personalized medicine system as an approach to discover that some tumors have unique pathologic and molecular characteristics that may warrant different treatment strategies. This research is based on the announcement of Qatar national genome project (to map the genome of the entire population of Qatar for delivering personalized medicine). The goal is develop a genomics data hub and establish an advanced big data analytic with modern data mining predictive modeling with high performance computing for memory-intensive genomic analysis/variant and data-intensive clinical analytic using petabytes of phenotype and Omics databases. Therefore, by understanding specific differences in tumor biology, researchers are identifying bio-markers for many tumor types, which are helping them to develop treatments targeting these underlying disease pathways. With these targeted therapies, clinicians can develop a more specific treatment strategy for some individuals that are potentially more effective based on the individual's tumor characteristics. The experimental and simulated genome-wide SNP data provided by the Genetic Analysis Workshop 16 and 17 will be utilized to investigate the new machine learning technique. This data afforded an opportunity to analyze the applicability and benefit of current machine learning methods, namely, penalized regression, ensemble learning methods, and network analyses resulted in several new findings while known and simulated genetic risk variants were also identified. The integrated strategies of both phenotype and Omics databases, implementation, and the learning processes are briefly proposed. The motivation of this research is to identify and discuss those GWAS challenges that will require breakthrough and innovative big data analytic and advanced predictive modeling frameworks to handle massive GWAS data towards personalized medicine at bedside. The ultimate goal is to deliver the right treatments to the right patients at the right time.

Abstract AIM: To generate soluble single chain variable fragments (ScFv) of monoclonal antibody recognizing the human breast cancer cell line (MCF7). METHODS: mRNA was isolated from the hybridoma cell line producing C3A8 monoclonal antibody and the cDNAs encoding variable domains of heavy and light chains (VH and VL) of the antibody were amplified Next, the ScFv DNA was ligated into the phagemid vector pCANTAB 5E and later transformed into E.coli TG1. Following transformation, the cells were then infected with M13KO7 helper. Then two biopanning step was carried ScFv-phages fusions antibodies were selected by ELISA. One selected clone was used to infect E.coli HB2151. The soluble ScFvs were then identified by Western blot, and their antigen-binding activity was assayed by ELISA. The scFv gene was then cloned into nova-blue host strain for cloning purposes and then into origami DE host strain for further characterization. The purified single-chain antibody expressed in origami DE3 was purified using Immobilized Metal Affinity Chromatography (IMAC). The purified scFv protein was characterized using western blot, flow Cytometery and immunofluorescence tests. Bioinformatics tools were also used and databases to gain specific functional insights into scFv anti-MCF-7. Finally, the VH and VL chains models were evaluated. RESULTS: The VH, VL and ScFv DNAs were about 340 bp, 320 bp and 750 bp respectively. The strongest positive clone (B7) was used to proceed for larger scale antibody production. The DNA sequencing results show that the ScFv gene has a gene bank similarity of 99%. Further, immuno-fluorescence test clearly proved that the scFv recognized the MCF-7 antigen epitopes which is localized in MCF-7 nuclear. Also, 53% of the cells numbers were bound to scFv protein as measured by flow cytometery analysis. Finally, the predicted structures of heavy and light chains were connected with peptide linker to build the full scFv protein structure. CONCLUSION: The soluble ScFv of is successfully produced, which not only provides a possible novel targeting vehicle for in vivo and in vitro study on associated cancers, but also offers the antibody a stable genetic source.

Description: Clinical nursing practice is on the cusp of significant and unrelenting change amid globalization, austerity measures, and technological advancements as the world moves out of the industrial age into the knowledge age. With advances in technology, theory, and research, the potential changes to future nursing practice are unlimited. Issues such as telehealth, nanotechnology, and globalization are but a few of the major trends influencing healthcare and nursing practice. As these trends become reality and the need for nurses to transition their practice to fit with the changing world, understanding how nurses change their practice becomes crucial. Aim: My purpose in this presentation is to argue the need for a grounded theory study to better understand the process of nursing practice change. Understanding the process nurses undertake to change their practice can help to identify strategies to facilitate change in a safe and effective method. The connection between practice change and factors that influence that change is not a new concept. The literature is replete with discussion about barriers and facilitators to Evidence-Based Practice (EBP), Research Utilization (RU), Quality Improvement (QI), Theory-Based Practice (TBP), yet the success of these approaches at influencing nursing practice change has been less than optimal. I believe one factor contributing to this lack of success is our deficiency in understanding the process nurses undertake to any of these approaches into her/his practice.

Abstract: Over the past few years, tissue engineering in dentistry has achieved relevant results. Several methods have been described to enhance tissue engineering and low intensity pulsed ultrasound (LIPUS) has shown to play an important role in tissue formation and regeneration. (LIPUS) can accelerate bone fracture healing and osteogenic differentiation. The aim of this study was to evaluate the effect of local use of autologous osteogenically induced gingival stem cells (OIGSCs) and LIPUS on the healing or periodontal defects in beagle dogs. We hypothesized that the use of OIGSCs and LIPUS can enhance regeneration of periodontal defect. Eight adolescent beagle dogs were used and periodontal defects were established in their third and fourth premolars. GSCs were isolated from connective tissue of interdental papilla of each dog, characterized by flowcytometry for stem cell markers and were differentiated using osteogenic medium to produce osteogenically induced gingival cells (OIGCs). Premolars were randomly divided into 3 groups. 1) Negative control collagen only 2) OIGSCs loaded in collagen scaffold 3) and OIGSCs loaded in collagen scaffold in addition to LIPUS application. LIPUS was applied to the right side of each animal for 20 minutes using a 2.5 cm transducer. The animals were sedated prior to each LIPUS application using Isoflurane inhalation anesthesia. Clinical assessments include pocket depth (PD) and attachment level (AL) and the depths of boney defect were used to evaluate the outcome of tissue regeneration. Results: There was statistically significant difference between OIGSCs and control group (P = 0.036) and between OIGSCs+ LIPUS and control group (P= 0.023) in furcation healing. However, there was no statistically significant difference in periodontal defect healing between the OIGSCs and OIGSCs +LIPUS groups with regards to PD, AL and boney defect depth. Conclusion: OIGSCs produced furcation regeneration with or without the use of LIPUS.