Qatar Foundation Annual Research Forum Volume 2013 Issue 1

Patient engagement in their care process, vis-à-vis self-management programs is an important element of the patient's longitudinal care plan, where the patient is encouraged and expected to achieve self-efficacy in the self-management of the disease through a regime of educational and behavioral modification strategies. To ensure the effectiveness of self-management programs, it is important that the proposed self-management interventions are (a) personalized to the unique needs and constraints of the patient; (b) based on sound theoretical health models; (c) based on validated health and behavior assessment tools to determine the patient's physical and behavioral dispositions; and (d) readily accessible to the patient through a ubiquitous medium, such as smart phones or the web. In this paper, we present a novel personalized self-management framework that delivers personalized health educational interventions to empower, educate and engage patients/individuals through self-observation, barrier identification, goal setting and action planning to achieve behavioral self-efficacy and self-regulation so that individuals can self-manage their condition. Our personalized self-management framework is guided by Social Cognition Theory, whereby have ensured that self-management programs for chronic disease management not just focus on changing the patient's awareness of the disease, rather they focus on enhancing the ability of the patient to make the right choices to achieve effective disease management. We present a three-stage personalized self-management framework that comprises: Stage 1: A high-level characterization of an individual with respect to a specific health outcome using validated assessment tools; Stage 2: A behavioral categorization of the individual based on his/her levels of self-efficacy, motivation and self-regulation, etc.; Stage 3: Use the personalized profile of the individual to tailor generic educational and self-management to develop a personalized self-management program that comprises personalized strategies to counter the challenges and barriers faced by the individual to achieving positive self-efficacy and self-regulation which in turn will lead to positive health outcomes. Our personalized self-management framework features (a) a novel self-management oriented individual profiling mechanism that takes into account both the health and psychosocial characteristics of an individual to generate his/her holistic profile; (b) a semantic web based knowledge model that captures the theoretical foundations of the SCT in terms of a Self-Management Program Personalization (SPP) ontology; (c) a semantic web based personalization tool that uses a logic-based execution engine that reasons over the SPP ontology, based on an individual's profile, to generate personalized self-management interventions; and (d) a mobile messaging platform to deliver the personalized self-management interventions and to monitor the patient's compliance using smart phones. We take a semantic web approach in designing the personalization approach whereby we have developed the SPP ontology to (a) model the theoretical framework of SCT in terms of SCT concepts; (b) model health assessment tools; (c) model the personalization rules that integrate the health and SCT models with the educational messages to generate a personalized self-management program. We have demonstrated the novel integration of health models, educational content and behavior change strategies to design self-management programs for cardiac risk factors, where the program is delivered through a mobile app.

Background: People with diabetes are at risk of developing foot ulcers (DFUs), which accounts for significant morbidity and mortality. Damage caused from repeated foot loading and temperature changes during walking may go undetected by patients due to loss of lower-extremity sensations (peripheral neuropathy). Clinical assessment on the other hand needs considerable time. Therefore, a feasible assessment of pressure and temperature is vital to measure pre-ulcerative inflammation and predict DFUs. Current study is aimed to validate effectiveness of an innovative smart textile based on fiber optics which allows measuring simultaneously plantar pressure, plantar temperature, and lower extremity joint angles. Methods: The proposed technology is based on optical fiber glass that propagates of light. Multiple sensors were juxtaposed on the length of the fiber which was integrated in a comfortable sock -SmartSox (Novinoor LLC, IL, USA) - Figure 1. Based on changes in wavelength of light, mechanical changes in environment including changes in temperature, pressure and joint angles are measured. The sensors were integrated in anatomical regions of interest including heel, mid-foot, 1st and 5th metatarsal heads, and under and over big toe. The later sensor allows measuring hallux range of motion in addition to temperature and pressure. To validate the accuracy of the designed prototype, 21 volunteers diagnosed with diabetes were recruited. Subjects were asked to walk a predetermined route of 200 steps in their normal or prescribed shoes. Plantar foot thermal images were taken by a thermal camera system twice: once at baseline after a five-minute temperature acclimation period and once after each walking trial. To determine the number of steps taken, subjects were asked to wear a gait analyzer system (LEGSys™, Biosensics LLC, MA, USA) in addition to SmartSox. To validate the accuracy of plantar pressure, a computerized pressure insole, F-scan (TekScan® Inc, MA, USA) was used as a gold standard. Results: Twenty-one patients with diabetes were recruited (Age: 57.8±7.9 years, BMI: 31.6±8.0 kg/m2, VPT: 26.8±15 volt, 68% diagnosed with peripheral neuropathy). All subjects perceived the SmartSox as comfortable and no problems were observed during walking assessment. Some fibers were however broken during wearing the socks in particular for patients who worn removable cast walker. A significant correlation was observed between the pressure profile measured by SmartSox and F-Scan under different anatomical regions of interest (r>0.6, p<0.05). A similar agreement between SmartSox and thermal camera was observed for changes in plantar temperature during walking. Conclusions: This study demonstrates the proofs of concept of an innovative smart textile for assessing simultaneously the key parameters associated with risk of foot ulcer in patients with diabetes. Fragility of the current prototype is considered as one of its major weakness for routine clinical assessment and thus further improvement in design of fiber and sock is needed. Additional study is required to address whether the measured parameters can be used to predict and better management of diabetic foot ulcer.

Modulation of calcium homeostasis plays a key role in regulating fundamental cellular processes, including gene transcription, cell proliferation, differentiation, and apoptosis. Store-operated calcium entry (SOCE) is the major pathway in non-excitable cells for extracellular Ca2+ influx across the plasma membrane and is regulated by the content of the intracellular Ca2+ stores. Following store depletion, the Ca2+ sensor in ER, stromal interaction molecule 1 (STIM1) clusters in ER regions close to the plasma membrane and recruits Orai1, which is a Ca2+ selective channel resulting in SOCE. Despite the significant knowledge in understanding STIM1 subcellular distribution dynamics, little is known about the trafficking of Orai1. Our laboratory previously showed that in Xenopus leavis oocytes Orai1 shuttles between plasma membrane and endosomal compartments, and that it internalizes during meiosis. However, the subcellular distribution and trafficking of Orai1 in mammalian cells is not fully understood. In this study we show that at steady state around 46% of Orai1 is on the surface of the plasma membrane of CHO cells, while the remaining 54% localizes intracellularly, suggesting that Orai1 constitutively recycles between the two compartments. Our time lapse imaging shows continuous shuttling of Orai1 to and from the PM with an exocytosis rate T1/2 of around 5 minutes. We also measured the endocytic rate of Orai1 at 5% min-1. To identify the domain within Orai1 required for its trafficking, we generated deletions of either the N- or C-terminus of Orai1. Deletion of N-terminus does not affect Orai1 trafficking to the cell membrane in CHO cells. In contrast deletion of the whole C-terminus (257-301) significantly altered Orai1 trafficking to the cell membrane. Deletion of amino acids 285-301 of Orai1 C-terminus does not have any impact on Orai1 trafficking to PM, suggesting that the information necessary for Orai1 trafficking to PM is located in the segment 257-285 of Orai1 C-terminus. In this study we determined, for the first time, the percentage of Orai1 on PM of mammalian cells and showed through time lapse-imaging that it constitutively recycles between PM and intracellular compartments. Our results also suggest, for the first time, that the amino acid region (257-285) of Orai1 C-terminus is essential for its plasma membrane trafficking.

Gastric bypass surgery is the most effective treatment for obesity, achieving both sustained weight loss and improved insulin sensitivity. However, following Roux-en-Y gastric bypass surgery (RYGB) some patients develop debilitating nausea and vomiting (N/V) persisting for years. The aim of this study was to determine if the N/V following RYGB is due to elevated systemic GLP-1 levels and whether GLP-1 directly mediates secretion of adipokines, such as leptin. 42 female non-diabetic subjects were studied in the fasting and post-prandial state and divided into 5 groups according to BMI and presence of N/V post-operatively. The effect of GLP-1 on adipose tissue leptin secretion in vitro was measured. Subjects with N/V post-RYGB surgery had significantly elevated fasting GLP-1 levels compared to post-operative subjects without N/V symptoms, and to morbidly obese (MO), obese and lean subjects not undergoing surgery. However, weight loss, as well as systemic levels of glucose, insulin and post-prandial GLP-1 was similar in all post-operative subjects. Despite similar BMI (P = 0.86) and fasting adiponectin levels, leptin was significantly lower in subjects with N/V compared to those without N/V (P = 0.04). Leptin secretion from subcutaneous adipose tissue in vitro was significantly inhibited by GLP-1 (0.1-1.0 nM; n = 6). Persistent N/V following RYGB surgery is associated with elevated fasting GLP-1 and lower leptin levels, perhaps as a consequence of GLP-1 mediated inhibition of leptin secretion from adipose tissue. GLP-1 antagonists may alleviate these symptoms, but, adverse effects on weight maintenance and insulin sensitivity need to be considered.

Epithelial ovarian carcinoma (EOC) is an aggressive neoplasm that mainly metastasizes to organs of the peritoneal cavity. This event is mediated by molecular mechanisms that remain elusive. Cell adhesion molecules play a key role in tumor invasion, metastasis and drug resistance. The conventional in vitro two-dimensional cell culture models are not sufficient to explain the exact mechanism behind tumor invasion, migration and anoikis resistance events. The objective of the present study is to analyze the role of cell adhesion molecules in tumor invasion metastasis and drug resistance using multiple phenotyping approach. Five ovarian cancer cell lines PA1, SW626, CAOV3, SKOV3 and OVCAR3 were selected for the study. Cell line phenotyping was conducted using cultured cells in an anchorage independent method that utilized an ultra low attachment plate for mimicking anoikis resistance in vitro. Second type phenotyping was done for sorting highly invasive phenotype by selecting cells that can pass through Boyden chamber (8 micron pore size upper chamber membrane). Development of drug-resistant cell lines were achieved by growing cells in culture media containing standard chemotherapeutic agents such as Taxol and Carboplatin. Cells were selected according to their ability to attach different ECM and cell adhesion molecule coated chambers. A real time PCR array of 29 genes involved in cell adhesion, drug resistance and EMT (epithelial-mesenchymal transition) were also analyzed and gene expression analysis conducted. The invasive potential of PA1 (teratocarcinoma) seems to be higher than other cell lines followed by SKOV3 cells. PA1 cells form embryoid bodies while culturing in in anchorage independent condition and exhibit an elevated level of expression of myc and TGF beta. Cell viability assay also shows that PA1 is the most sensitive cell line against carboplatin and taxol. Gene expression levels of cell adhesion molecules were altered among the phenotypes. Anoikis resistant cells show altered levels of expression in EPCAM, Collagen 6, CD24, vimentin and N-cadherin. These results suggest the cell lines are heterogeneous in nature and three dimensional culture model mimics the in vivo tumor model for anoikis resistance and EMT. To conclude, this study shows selection of various phenotypes of heterogeneous cancer cell lines can help decipher the role of cell adhesion molecules in ovarian cancer invasion and drug resistance. These experiments will give an overall view of the role of cell adhesion molecules in different ovarian cancer types in vitro.

Asperger syndrome is one of the Autism Spectrum Disorders (ASD's), characterized by significant difficulties in social interaction and nonverbal communication, alongside restricted and repetitive patterns of behavior and interests. ASD's have a strong and complex genetic basis that cannot be distinguished by the clinical presentation. A South African family with an affected father and three affected sons all with characteristics of Asperger syndrome including repetitive routine physical gestures and Sensory Processing Disorder was studied for the possible identification of the responsible genetic factor(s). Due to the significant proof of heritability and the extreme heterogeneity of Asperger syndrome, next generation sequencing was performed on all members of this family to extrapolate monoallelic variations in the affected father that have been inherited by all three of the affected sons probably through an autosomal dominant pattern of inheritance. These variations, validated by Sanger sequencing, were analyzed, prioritizing significant changes in the encoded protein. Variants that segregate with the affected individuals include one deletion in C17orf80, an unidentified protein expressed in the brain, (c.1745_1748delGTAA), three missense mutations that change highly conserved amino acids in PARK2, which codes for a component of a multiprotein E3 ubiquitin ligase complex that targets proteins for degradation also known to cause juvenile Parkinson disease (c.110 C>T, p.Pro37Leu), FAT1, member of a large cadherin family required for cell-cell association and actin organization, (c.2563 C>A p.Gly855Arg), and OR4C6, an olfactory receptor protein coding gene, (c.293 A>C p.Gln98Pro) and one nonsense mutation introducing a premature stop codon in HYAL4, a hyaluronidase that intracellularly degrades hyaluronan, one of the major glycosaminoglycans in the extracellular matrix (c.628 C>T p.Arg210STOP). The direct link of these previously reported variants to Asperger syndrome is yet unknown, however some of these genes such as PARK2, HYAL4 and FAT1 have previously been reported to be in involved in brain function and development, indicating a possible role in the onset of Asperger syndrome.

This study evaluated the effectiveness of paravertebral block as alternative anesthetic technique for extracorporeal shock wave lithotripsy (ESWL) procedure. Fifty patients with renal stones, aged 20 to 60 years, were randomly allocated into two groups; twenty five patients in group P; received unilateral paravertebral block from T8 through L1 with injection of 5ml 0.5%bupivacaine and 25 patients in group L; received local infiltration by bupivacaine 0.25%(2mg/kg) into the 30 cm2 area after localizing the stones site, 10 minutes before the session. 10 mm visual analogue scale (VAS) was used to evaluate pain every 10 minutes during the session. At the end of the procedure, total doses of rescue analgesia, the number of shockwaves, their power and the total duration of shockwave treatment were recorded. After completion of the procedure the patient was assessed for pain and nausea in the post anesthesia Care Unit (PACU) using the Visual Analog Scale. Patient's satisfaction and time needed to discharge patients to home also were recorded. Time to do the anesthetic technique was significantly higher (p<0.001) in group-P than group-L, it was 12.7±2.3 minutes versus 6.9±1.9 minutes respectively, intraoperative rescue analgesia by fentanyl was lesser (P<0.001) in group-P than group-L, 26.7 ± 6.32mcg versus 78.6±5.41mcg, respectively, also time interval between ends of the procedure till discharge to home was significantly higher (P<0.001) in group-P than group-L, it was 99±17 minuets versus 133±31minuits respectively. VAS was not significant difference between both groups either intraoperative or postoperative in first hour. Patient's satisfaction was significantly higher (P<0.05) in group-P than group-L, it was 8.8±1.1 versus 6.1±0.6, respectively. Adverse events were lesser, but not significant in group-P than in group-L. Two patients (8%) in group-L and one patient (4%) in the group-P experienced episodes of postoperative nausea and vomiting (PONV). Paravertebral block is effective alternative anesthesia for outpatient lithotripsy; multiple level paravertebral blocks provide an optimal anesthetic condition, with acceptable adverse events for ESWL. And providing proper analgesia during the procedure and in first hour after finishing of the procedure, early discharge to home and providing better patient's satisfactions.

Abstract: Background: While promising, cationic lipid-mediated gene delivery can still benefit from improvements in lipid design and lipid-DNA (lipoplex) formulation. The putative mechanism of cellular lipoplex uptake is believed to occur by endocytosis, where the key influential factors are lipoplex size and morphology; lamellar and inverted hexagonal. Lamellar lipoplexes offer superior protection to the DNA cargo, while the inverted hexagonal phase best facilitates endosomal escape. Ideally, the initial lipoplex packaging would have the lamellar phase upon uptake, followed by a phase transition to hexagonal, facilitating cargo release into the cytosol. The cationic lipid structure defines its molecular packing parameter, S, which in turn controls the lipid phase transition. A molar weighted average packing parameter (Smix) for the overall cationic and neutral co-lipid mixture within a lipoplex formulation is predictive of a lamellar (S&lt;1) or hexagonal (S&gt;1) phase lipoplex. Objectives: Pyridinium-based cationic lipids represent a class of non-viral vectors that have shown promise in gene delivery. The objective of this study was to test the influence of lipid shape on lipoplex phase structure through small-angle x-ray diffraction (SAXD), and correlate shape with transfection efficiency. Lipoplexes co-formulated with pyridinium lipid, (16:0)(11:1) having a calculated shape parameter, S, of 1.08 and the commercial cationic vector, EPC (S=0.94) were predicted to undergo a packing transition from lamellar to hexagonal as the ratio of (16:0)(11:1) / EPC is increased. A fixed amount of neutral co-lipid was employed (1,2-dioleoyl-sn-glycero-3-phosphoethanolamine, DOPE: S=1.01; or cholesterol, Chol: S=1.20). Given that cholesterol is a higher-S lipid than DOPE, the lipoplex phase transition from lamellar to hexagonal was anticipated to occur at a lower ratio of (16:0)(11:1) to EPC. Methods: Liposomes were prepared from pyridinium-based cationic lipids in combination with EPC and co-lipid, DOPE or cholesterol. Lipoplexes were then formulated by incubating the liposomes with plasmid DNA at various N/P (+/-) molar charge ratios, and subsequently characterized by gel retardation, DNAse I degradation, biocompatibility and β-galactosidase (β-gal) transfection assays using Chinese Hamster Ovarian (CHO-K1) cells. Lastly, lipoplexes at N/P molar charge ratio 3 (only) were analyzed by SAXS at the synchrotron in Grenoble, France. Results: The SAXD results revealed that the (16:0)(11:1)/EPC/DOPE-DNA lipoplex formulations underwent a lamellar to hexagonal packing transition when the (16:0)(11:1)/EPC molar ratio was increased from 1:2 to 1:1, where the Smix increased from 1.01 to 1.04. However, (16:0)(11:1)/EPC/Chol-DNA lipoplex formulations underwent a lamellar to hexagonal transition when the (16:0)(11:1)/EPC molar ratio increased from 1:1 to 2:1; when Smix increased from 1.11 to 1.13. For both DOPE and Chol containing lipoplexes, the greatest transfection was found at (16:0)(11:1) to EPC ratios below 2:1, at an N/P molar charge ratio of 3. Conclusion: The lamellar to hexagonal packing transition, as determined by SAXD, for the lipid-DNA lipoplexes composed of the (16:0)(11:1)/EPC mixture occurred as predicted by our Smix calculations when DOPE was employed as co-lipid. The same was not observed when cholesterol was employed co-lipid. Finally, superior lipoplex transfection correlated with lamellar packing.

Introduction: Mild Brain Injury or concussion is frequently observed during contact sports such as football and soccer. If it remains undetected, a repeat concussion can lead to long term consequences because of the vulnerable brain tissue damage. Balance Error Scoring System (BESS) test is a widely used test to detect concussion. It requires the athlete with suspected concussion to maintain his/her position in three different stances, that are, both legs, single leg and tandem, and has a total score ranging from 0 to 30. The recommended way of performing this test is to do it barefoot in clinical or semi-clinical settings. Such conditions are however difficult to achieve during an ongoing match, and athletes would like to be near the field with their cleats on - situations often found on soccer fields in eastern countries. Objective: Therefore, we aimed to assess the reliability of BESS test in different field conditions. This research is in line with Qatar National Research Strategy 2012 pillars, H.E.1.9 (prevention of brain Injury) and H.E 1.10 (control of sports injuries). Methods: This study was conducted under the auspices of McGill Sports Emergency Medicine Clinic. Athletes from soccer and football teams were approached on the field during practice games. After informed consents, they performed BESS test in three conditions, that were, barefoot, on turf with cleats and on hard surface with cleats. Each athlete was rated by three observers independently of each other. We computed mean difference in total BESS scores with 95% confidence intervals (95%CI). Comparison of total BESS scores under different conditions as well as inter-observer reliability was assessed using intraclass correlation coefficient (ICC).. Results: We recruited 49 athletes from football (n=39) and soccer (n=10) teams in this study. Thirty nine of them were male, 10 were females. Average age was 21.1 years (standard deviation [SD]=1.9). We found that total BESS scores were significantly different (P&lt;0.001) between barefoot and the two conditions with cleats-on: 2.2 (95%CI=1.6, 2.8) for turf and 2.0 (95%CI=1.4, 2.6) for hard surface. Concordances of barefoot with turf (ICC=0.47, P=0.02) and hard surface (ICC=0.51, P=0.01) conditions were moderate. A moderate to high inter-observer reliability (0.60≥ICC≤0.75) was observed for BESS test under three conditions. Conclusion: These findings show that BESS test has a fair reliability under different conditions, and may be useful in screening concussion on the field. However, cut-off of BESS should be reduced by 1 to 2 points if it is applied on the field with cleats.