Qatar Foundation Annual Research Conference Proceedings Volume 2014 Issue 1
- تاريخ المؤتمر: 18-19 Nov 2014
- الموقع: Qatar National Convention Center (QNCC), Doha, Qatar
- رقم المجلد: 2014
- المنشور: ١٨ نوفمبر ٢٠١٤
201 - 250 of 480 نتائج
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Lower Extremity Blood Flow Change From A Low Impact Virtual Reality Training In Diabetes
Background: Inadequate blood flow can increase risk of foot ulceration in diabetics. Exercise training can improve lower extremity blood flow in diabetes, however, the intensity of exercise should be taken into account for this population is they are at high risk for developing foot ulcers. Aim: The aim of the present study is to explore the immediate effect of a tailored low impact virtual reality exercise training on lower extremity blood flow in patients with diabetic peripheral neuropathy. We hypothesize that tailored exercise specifically designed for diabetes will induce sufficient muscle activation in order to improve blood flow. Methods: Eight patients (Age: 64.3±10.2 years, BMI 32.6±6.3) with diagnosed DPN (HbA1c: 7.3±1.2) participated in a single 45 minute session for low-impact virtual reality based balance training exercise. Changes in blood flow were quantified by Ankle Brachial Index (ABI) recorded pre- and post-training. Systolic blood pressures were determined at the brachial, posterior tibial, and dorsalis pedis arteries using a 10 MHz handheld Doppler. Results: Non-parametric Wilcoxon Signed Rank test revealed a strong but non-significant reduction in ABI from single session virtual reality training. The values dropped from 1.28 to 1.22 (4.7% reduction, p = 0.173). Conclusion: The initial results show that a tailored low impact exercises for diabetes can improve blood flow, which may reduce the risk for diabetic foot ulceration. Future studies should confirm these findings in a larger pool of patients with 8-12 week exercise training in order to explore the long term effect on exercise on blood flow. Funding Source: QNRF NPRP 4-1025-3-276
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Women Versus Men With Depression Among Patients Admitted With Cardiac Events: The Impact Of Age And Socioeconomic Factors-Findings From A Middle-Eastern Contemporary Prospective Study
Introduction: Depression is associated with cardiovascular diseases. Early detection and intervention for depression among cardiovascular (CV) patients can reduce morbidity and mortality rates. Understanding age and gender differences is necessary to adequately address the complex nature of depression as co-morbidity among Arab CV patients in the Middle East region. Objectives: To evaluate the prevalence of depression, to find ways to manage depression among male and female Arab CV patients. Methods: A cross-sectional survey was conducted with 686 (69%) male and 314 (31%) females Arab CV patients between January, 2013 and March, 2014 at the Heart Hospital in Qatar. Inclusion criteria were ≥20 years of age, agreeing to participate in the study (98% response rate), and having final confirmation of acute cardiac conditions. Face-to-face interviews were conducted using structured survey questionnaires which included an Arabic demographic questionnaire and the Arabic version of the Beck Depression Inventory 2nd Edition. Results: 80% of the patients had no depressive symptoms, 20% of the patients suffered `mild mood disturbance` and clinical depression. Almost twice as many females (28%) than males (17%) were assessed having depression. Chi-square tests indicated that age and socioeconomic factors such as nationality, marital status, monthly income, employment, occupation, financial stress and support were significantly related to gender and depression (p<0.001 for all). Conclusion: Routine systematic screening for depression is recommended for all CV patients, especially for older women group. Although health care services in Qatar are heavily subsidized, socioeconomic factors influence male and female CV patients' mental health differently, thus their CV conditions and outcomes. To effectively manage depression among Arab CV patients, health care providers should be aware and integrate gender differences approach into their clinical practice.
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Validation Of An Optical Fiber Based Smart Textile: A Clinical Tool For Predicting Diabetic Foot Ulceration
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
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The Metabolic Syndrome Prevalence And Comparison Of The Different Subtypes Of Obesity Among Qatari Men And Women: The Qatar Biobank Project.
Background: Qatar is facing an epidemic of metabolic diseases such as obesity, and insulin resistant due to the rapid lifestyle and nutrition transition in the last few decades. Previous epidemiological studies have identified high prevalence of the metabolic syndrome (MetS) and obesity among Qataris. However, these studies are limited. Although obesity is an established risk factor for the MetS, studies have identified a subtype of obese individuals with healthy metabolic profile. To our knowledge, there were no identified studies that looked into the different subgroups of obesity among Qataris. Aims: This study aims to estimate the prevalence of the MetS. Moreover, to identify the various obesity subtypes and assess the difference between them regarding selected determinants. Methods: This research was conducted among 499 Qatari men and women aged ≥18 years old of the Qatar Biobank pilot phase cohort. The MetS prevalence was identified using the National Cholesterol Education Program's Adult Treatment Panel III (NCEP ATPIII) and the International Diabetes Federation (IDF) criteria. Four different subgroups of obesity were identified; these were Metabolic Healthy Normal Weight (MHN), Metabolic Abnormal Normal Weight (MAN), Metabolic Healthy Obese (MHO), and Metabolic Abnormal Obese (MAO). The classification was based on the body mass index (BMI) and the absence or presence of at least one metabolic risk factor of the MetS-NCEP ATPIII definition. Comparisons between the groups were conducted in relation to demographic, anthropometrics, lifestyle and biochemical characteristics using General linear models with post hoc Bonferroni correction. All models were adjusted for age and sex. Results: The MetS prevalence was 24.6% based on the NCEP-ATPIII definition and 27.1% based on the IDF definition. Among the studied cohort, 23.1% were classified as MHN, 13.2% were MAN, 13.6% were MHO and 50.1% were MAO. Compared to MHN, MHO subjects were characterized with significantly lower concentration of high-density lipoprotein (HDL), and two-fold higher insulin and c-peptide. On the other hand, MAO subjects were characterized with significantly lower physical activity and HDL levels compared to MHN, moreover, higher alkaline phosphatase, triglyceride, diabetes and blood pressure variables were found. When comparison between MHO and MAO was made, the latter had significantly lower HDL, and significantly higher triglycerides, waist circumference, waist-hip ratio, blood pressure and diabetes variables. Conclusions: The estimated MetS prevalence was 24.6% (NCEP-ATPIII) and 27.1% (IDF). About half of the studied individuals were classified as MAO. Although MHO and MAO were both characterized with a BMI ≥30 kg/m2, MHO subjects had significant smaller waist circumferences and waist-hip ratio. Additionally, MHO subjects had significantly healthier triglycerides, HDL, blood pressure and diabetes variables profiles. Due to the different characteristics, diverse therapeutics and interventions should be made for each subgroup of obesity. Keywords: Metabolic Syndrome, Obesity, healthy obese, Qatar Biobank.
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EDHF Contribution To Microvascular Dilatation Is Not Linked To Endothelial Dysfunction In Morbidly Obese Qataris
المؤلفون: Nelson Orie, Aysha Bakhamis, Moataz Bashah, Mohammed Alsayrafi and Vidya Mohamed AliObesity 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
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Rhythmic Succession Of Molecular Functions
المؤلفون: Abeer Fadda and Andrey PtitsynPeriodicity in cellular processes is arguably the most underestimated phenomenon in molecular biology. We previously demonstrated the circadian oscillation of baseline expression in eukaryotic genes showing that cycling is a basic property of all genes and not a function of only a subset (10-15%). We further explore the periodicity of cellular processes in this study by asking the question of whether the circadian cascade of gene expression is reflected by a cascade of molecular functions. We do this by reanalyzing a set of time-course gene expression data from heart tissue of wild type and CLOCK-mutant mice. The analysis of sets of oscillating genes that peak at different times during the day, demonstrate that some molecular functions and processes are more active at certain times than others. Comparison between wild type and CLOCK-mutant tissue samples shows the difference not in the overall number of oscillating genes, but rather in coordination of molecular functions with time. These findings will impact the choice of background controls for future expression studies, and improve the models for biological pathways to name a few of its implications.
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Patient-Specific Seizure Onset Detection Via Neural Synchrony
المؤلفون: Marwa Qaraqe, Muhammad Ismail and Erchin SerpedinBackground & 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.
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The Combined Use Of Thermachoice Ablation And Mirena In The Management Of Heavy Menstrual Bleeding
المؤلفون: Alex Tan, Syamali Basak and Imad AbukhalilIntroduction Heavy menstrual bleeding (HMB) commonly affects 1.5 million women in the UK, thereby prompting 1 in 20 women of reproductive age to seek treatment to improve their quality of life. The NICE guidelines recommend the use of Mirena in treating benign HMB, followed by endometrial destruction, if medical treatments fail to resolve symptoms. Several studies have shown the efficacy and cost-effectiveness of using Mirena, endometrial ablation and hysterectomy individually, but evidence on the efficacy of combining endometrial ablation and Mirena compared with other treatments remains inadequate. The objective of this study was to evaluate the efficacy and outcomes of women with HMB via combined use of Thermachoice ablation and Mirena. Methods Forty-nine patients with persistent HMB following initial treatment involving either Mirena, Thermachoice ablation or medical treatments were randomly selected from the sample population between 2005 to 2011 and consented to have combined Thermachoice ablation and Mirena. Our primary outcome measures were satisfaction levels 6 months post-treatment, amenorrhoea rates and rates of subsequent hysterectomy. Results There were similar number of patients in the each age group, including aged 35 to 40 (15 patients), 41 to 45 (17 patients) and 46 to 50 (15 patients), except for 2 patients aged beyond 50. 33 of 49 patients had at least two live births and BMI of above 30 respectively. 36 (73%) patients had normal pelvic scan findings, 10 patients had abnormal findings and subsequent histology specimen excluded hyperplasia, atypia and malignancy, and 3 patients had inconclusive findings due to high BMI. Out of 49 patients, 18 (37%) had Mirena, 2 (4%) had Thermachoice ablation and 29 (59%) had medical treatments after declining Mirena initially. Following initial treatment, 16 patients with Mirena and all patients who had ablation and medical treatments (96%) developed initial spotting that persisted to severe loss. Following subsequent counselling and combined treatment of Thermachoice ablation and Mirena, 38 (78%) patients were very satisfied with their symptoms (3 of 3 satisfaction ratings), 36 (73%) patients had high amenorrhoea rates and only 11 (22%) patients had persistent spotting that led to a hysterectomy. No other serious complications occurred in these patients following subsequent treatment. Conclusion The combined use of Thermachoice ablation and Mirena in treating HMB remains justified based on the overall outcomes and a large prospective study is required to compare its efficacy with other treatments. References 1. Vaughan D, Byrne P. J Obstet Gynaecol 2012 May; 32(4): 372-4.
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Qatar Cultivated Vegetables Enhance Resistance To Protein Modifications Induced By Altered Redox Homeostasis
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
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Follicular Dendritic Cell Sarcoma: Cytogenetics And Pathological Findings
المؤلفون: Udayakumar Muthappa AchandiraFollicular Dendritic Cell Sarcoma: Cytogenetics and Pathological Findings Abstract Follicular Dendritic Cell Sarcoma (FDCS) is a rare neoplasm with a non-specific and insidious presentation that is further complicated by difficult diagnostic and therapeutic assessment. It has a low to intermediate risk of recurrence or metastasis. Unlike other soft tissue sarcomas, cytogenetic studies are very limited in FDCS. Although no specific chromosomal marker is established yet, complex aberrations, different ploidy types are documented. We present a case with ultra-structural, immunophenotypical and histological features. The karyotypic findings were different from those reported in literature. Keywords: Chromosomal aberrations; Cytogenetics; FDCS; Fine needle aspiration;Follicular dendritic cell sarcoma;karyotype
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Targeting The Warburg Effect Via Polyamine Metabolism For Prostate Cancer Treatment And Improved PET Imaging
المؤلفون: Salim Merali and Magid Abu-gharbiaAlthough 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.
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A Mutation In MYO1A Causes Autosomal Recessive Autism Spectrum Disease
A consanguineous family of Pakistani ethnicity with two female siblings (22 and 19 years of age), affected by a autosomal recessive Autism Spectrum Disease, was studied by homozygosity mapping and whole Exome Next Generation Sequencing [NGS] of the two affected siblings and one parent to identify the responsible gene and mutation. The disorder is marked by intellectual disability, speech and motor delay, congenital malformations and possibly autism spectrum disorder (ASD). The malformations include microcephaly, microphthalmia, micrognathia and arachnodactyly with hyperextensibility and persistent fetal pads in fingers and toes. The offending gene was mapped to five possible homozygous genomic regions [[6q, 12q, 17p, 20p, 22q], as the family structure did not allow identification of a single interval with a significant LOD score. Comparative analyses of the NGS data for autosomal recessive inheritance and data mining for damaging variants within the homozygosity intervals identified a damaging homozygous c.C1675T / p.R559C mutation in the MYO1A gene, at 12q13.3. The mutation co-segregates with the disease phenotype within the family, is absent in known polymorphism databases and in 400 ethnically matched control chromosomes. Myosins are molecular motors that, upon interaction with actin filaments, utilize energy from ATP hydrolysis to generate mechanical force. The N-terminal motor domain contains both ATP-binding and actin-binding sequences. Following the motor domain is a light-chain-binding 'neck' region containing 1-6 copies of a repeat element, the IQ motif that serves as a binding site for calmodulin and other members of the EF-hand superfamily of calcium-binding proteins. The C terminus has a distinct tail domain that serves in dimerization, membrane binding, protein binding, and/or enzymatic activities and targets each myosin to its particular subcellular location. Heterozygous mutations in MYO1A have been found in patients with sensorineural hearing loss, speculated to cause autosomal dominant sensorineural hearing loss but co-segregation to the phenotype has never been demonstrated. Two rare heterozygous MYO1A mutations [c.G2021A / p.G674D and one in the 3′UTR] have been found in patients with autism but their clinical significance is unknown. The association of MYO1A to autosomal recessive ASD without deafness in this family, elevates the importance of MYO1A both as causative and contributive gene for Autism Spectrum Disease
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Pathophysiological Features Of Impaired Fasting Glucose (IFG) And Impaired Glucose Tolerance (IGT) In Arab Individuals
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
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Lattice Based Mispronunciation Detection For The Assessment Of The Childhood Apraxia Of Speech
المؤلفون: Mostafa Ali Shahin, Beena Ahmed and Kirrie BallardBackground and Objectives Childhood Apraxia of Speech (CAS) is a speech disorder characterized by articulation errors, i.e. the replacement of certain phonemes with alternatives. In previous work we proposed a simple method to evaluate the child's speech as correct or incorrect with an overall accuracy of 88.2%. In this work we present an enhanced method that increases the accuracy of the correct/incorrect evaluation to 92.7%, in addition to identifying the incorrect phonemes with an accuracy of 60%. Method The goal of the mispronunciation detection system is to compare each phoneme in the child's production to their given prompt and identify mispronunciations. Figure 1 shows the block diagram of the system, which uses a search lattice for each prompt in the child's speech therapy treatment protocol to identify errors made. Each prompt is transcribed as per the corresponding phoneme sequence using the CMU pronunciation dictionary and then passed to the lattice generator along with the expected mispronunciation rules to generate the search lattice. Mel Frequency Cepstral Coefficients (MFCC) are extracted from the speech signal with delta and acceleration to produce a 39- dimensional feature vector per frame. The extracted features are then fed to the speech recognizer along with the created lattice and the Hidden Markov Model (HMM) acoustic models to generate a sequence of phones from the child's utterance. An evaluation report is then generated by matching the recognized phoneme sequence with the correct phoneme sequence and specifying the errors made by the child. We use a search lattice with a specific number of alternative pronunciations for each phoneme; this limits the decoder search, making it faster and more accurate. Each phoneme in the correct phoneme sequence is compared with expected mispronunciation rules developed by a therapist after an assessment of 20 children with CAS; if a rule is matched, the pronunciation variants are added as alternative arcs to the current phoneme sequence. The mispronunciation rules depend on the type of the phoneme (consonant/vowel), the phoneme position in the word (Initial/Medial/Final) and the context of the phoneme. The lattice is then created using the matched rules as shown in Figure 2, where the garbage model absorbs any mispronounced phoneme not in the lattice. PA and PG are insertion penalties added to the alternative and the garbage arcs respectively so the decoder does not align the speech to the alternative error phonemes or the garbage node unless it is confident enough. Results The system overall system accuracy is 92.7% where the Correct Acceptance (CA) is 97.6% and the Correct Rejection (CR) is 83.1%. The system also detects phoneme errors made by the child with 60% accuracy. Conclusion In this paper we proposed a mispronunciation detection tool that can detect phonemes mispronounced by children with CAS and specify the errors made.
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Qatar Experience In Standard Breast Cancer Screening
المؤلفون: Hekmet Abubaker Bugrein and Salha BujassoumQatar 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.
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A Prototype Of Virtually Interactive Hand Activating Devise - Low Cost Portable Head Mounted System (VIHAD Plus) For Neurological Rehabilitation
المؤلفون: Ajimsha Mohammed Sharafudeen, John John Cabibihan, Uvais Ahmed Qidwai and Lina MajedABSTRACT Background and Purpose: Restoring function in individuals who have severe paralysis of the upper extremity secondary to stroke is challenging. Recent technologies have made it possible to use robotic devices as novel tools for assisting the therapists to provide safe and intensive rehabilitation with repeated motions. However, most of the training robots are types of Continuous Passive Motion (CPM) devices that produce slower and stereotyped movement patterns. Earlier works have shown that passive or slow movements do not significantly benefit motor improvement. Several studies reveal that even the use of ipsilateral electromyographic (EMG) pattern recognition approaches might not be practical to decode movement intention and, may negatively affect re-mapping of the neural pathways in the brain. To have a successful hand rehabilitation system, the system should be able to produce a wide variety of unpredicted and challenging movement patterns of various degrees of speed and range of motion with increasing complexity, sufficient enough to produce the necessary neurological plasticity of the affected brain. Current rehabilitation devices are not sufficient to produce such a range of complex activities which enables maximum neurological plasticity. Objective: In this work, we describe a prototype of the contralateral EMG-based Interactive Hand Activating Devise for Stroke (IHADS) system that can detect a hemiplegic person's intention for bilaterally executed hand activities using his/her surface EMG signals from the non-affected side (contralateral). Furthermore, this system can assist in bilateral hand activities through an exoskeleton attached to the hemiplegic upper extremity to initiate progressively challenging and unpredicted type of activities in a virtual reality (VR) world to obtain optimum functional recovery by inducing maximum neurological plasticity. Design: The IHADS system is made up of an embedded controller and a robotic exoskeleton, contralateral EMG sensors and a VR interface with a semi-immersed VR system, where the patient will be seeing progressively impulsive activities that would force the brain to activate the affected extremity to manipulate through the remaining neural networks and mirror neuronal system which in turn will optimize the neurological recovery. This means that the paralyzed arm will be following the motion of the healthy arm whose motion is picked up by the EMG sensors and are translated as actuation signals for the exoskeleton to execute virtually created challenging activities. Conclusion: Contralateral EMG-based 'IHADS' system is a unique, cost effective, highly innovative and portable robotic device. If incorporated into the stroke rehabilitation, this system will be capable of autonomous guidance through the use of real-time feedback from the contralateral upper limb, integrated via the VR interface and the hand activating device to make rehabilitation more intense, functional, motivating, and capable of inducing maximum neurological plasticity. Keywords: contralateral upper limb EMG, exoskeleton; robot; virtual reality, stroke rehabilitation
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Retinal Imaging: A Convenient Tool To Study Microvascular Responses To Cardiovascular Disease Risk Factors And To Predict Hypertension And Disease Development
المؤلفون: Patrick De Boever, Tijs Louwies, Eline Provost, Luc Int Panis, Arnout Standaert, Roel Smolders and Johan GeysenThe 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.
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A Pilot Study On Cognitive-behavioral Approach For Combatting Childhood Obesity In Qatar
Levels of overweight and obesity have reached alarming proportions in Qatar and other Gulf nations. In Qatar, the need to establish national strategies for the prevention and treatment of obesity was recognized in the National Health Strategy 2011-2016, which stresses the need for prevention. The treatment and prevention of childhood obesity is largely through lifestyle changes- encouraging health eating and physical activity. However, changing such behaviors is complex and requires integrated approaches to tackle such a multifaceted problem. In this study, we sought to implement and evaluate a novel weight management program for Qatari school children at the vulnerable age of 9-12 years. The program uses a cognitive-behavioral approach that involves developing social and emotional competences, promotion healthy dietary habits, and development of physical literacy, while enlisting family involvement in an attempt to maintain long term weight loss. This study also sought to take things further by integrating cutting edge insights from behavioral economics. The pilot study was conducted by an interdisciplinary team of collaborators from Qatar (Qatar University, Supreme Education Council, Aspire, Hamad Medical Corporation) and external partners (Imperial College and Leeds Metropolitan University/MoreLife, UK). The study was branded Agdar/أقدر and involved development of intervention materials, questionnaires, neuropsychological measures, and other study components such as training manuals and virtual world program. A sample of 100 overweight or obese Qatari children (age 9-12) from 5 independent schools participated in the intervention along with a control group. The intervention was conducted in Arabic and consisted of three phases (1) intensive weight loss camps, (2) after-school clubs as consolidation phase, and (3) a maintenance phase using virtual world program. Variables measured included anthropometric (BMI, Fat composition, and Waist circumference, and Blood pressure), Lifestyle (Physical Activity and Dietary questionnaires), and Psychometric assessments (Self-esteem, Subjective well-being, and Impulsivity, including heart monitoring). Overall, 100% of children who participated in the camp lost weight with a significant reduction BMI-SDS (p<0.001). The camp led to a significant improvement in subjective wellbeing in all five dimensions (p<0.01). The afterschool clubs showed a synergistic effect on children who lost weight at camp where further weight loss was achieved in the after school club phase. The overall percentage BMI-SDS reduction (-8.7%) was significantly higher than the required reduction for health benefits in both adolescents (-3%) and adults (-5%). Girls continued to outperform boys in terms of weight loss reduction and maintenance (-11.9 vs. -5.6%, respectively). This success occurred despite pre-camp surveys showing nearly two thirds of children not meeting their daily requirements of fruits and vegetables while consuming calorie-dense fast foods and over half of the participants not engaging in physical activity. Physiological parameters such as standard deviation of the heart beat-to-beat intervals, used as proxy for impulsivity, correlated significantly with BMI-SDS (p<0.035). Children with a higher BMI were less able to withhold their choices (proxy for impulsivity) and the physiological control of their heart rate correlated with this. These findings could help in segmentation of intervention participants into groups to which tailored programs could enhance impact on behavior change.
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Towards Efficient And Comprehensive Healthcare/Medical Monitoring System: Nano-Scale Body-Centric Networks (Nano-Health)
المؤلفون: Ke Yang, Nishtha Chopra, Qammer Hussain Abbasi, Khalid Qaraqe and Akram AlomainyA 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.
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Identification Of Proteins Involved In Orai1 Trafficking By Mass Spectrometry-based Approach
المؤلفون: Maya Dib, Rawad Hodeify and Khaled MachacaStore-operated calcium entry (SOCE) is a ubiquitous Ca2+ influx pathway essential for many physiological functions. Dysregulation of SOCE causes disruption in Ca2+ homeostasis leading to cellular pathology of several diseases. Orai1, a key regulator of SOCE, constitutively recycles at steady state in the frog oocyte and internalizes into an intracellular vesicular compartments during meiosis, leading to inactivation of SOCE. Previous data showed a role for Orai1 C-terminus in its internalization during meiosis. However, the minimal region required for Orai1 internalization at steady state and during meiosis is not known. In this study we investigate the molecular determinants of Orai1 trafficking in Xenopus oocytes by comparing the localization of multiple Orai1 C-terminal mutants 1-266, 1-275, and 1-285 intracellularly. Orai1 mutants 1-275 and 1-285 were both present in endosomes and were internalized during meiosis behaving similarly to WT. However, Orai1 1-266 was enriched at the plasma membrane and showed decreased localization to the endosomal compartment at steady state. Furthermore, in contrast to wild-type Orai1, it was not internalized during oocyte maturation. To identify proteins interacting with Orai1 and involved in its trafficking, we co-immunoprecipitated Orai1 complexes from oocytes expressing WT Orai1 and Orai1 1-266 and analyzed complexes by mass spectrometry. Our preliminary data identified several candidate proteins co-immunoprecipitating with WT Orai1 but not with Orai1 1-266 mutant. To summarize, our results show that the region of Orai1 containing residues 267-275 is likely essential for Orai1 localization to endosomes suggesting a role for this domain in Orai1 intracellular distribution. Mass spectrometry analysis on co-immunoprecipitation of Orai1 WT identified several candidates that are missing in Orai1 1-266 co-IPs, suggesting a role for these candidates in Orai1 trafficking.
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Differentially Expressed Circulating And Cell Associated MicroRNAs In The Investigation Of The Role Of Viral Infection In Type 1 Diabetes
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.
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Children With Type 1 Diabetes, Role Of Pro-inflammatory Cytokines As Disease Biomarkers
Background: Cytokines are important mediators of the inflammatory response in type 1 diabetes (T1D). We have previously observed pro-inflammatory cytokine profiles in children with islet autoimmunity. In the present study we examined the role of cytokine profiles as immunological biomarkers of T1D in a case-control study of 162 children at T1D onset and 164 community controls matched for age, sex and time. The diagnosis of T1D was based on presence of islet autoantibodies and clinical criteria. Methods: We measured 50 cytokines, chemokines and growth factors using the Millipore Cytokine Magnetic Bead Multiplex Assay. Heat maps and multivariate analyses were used assess how cytokine expression profiles may identify cases of T1D. Results: The levels of 30 cytokines, chemokines and growth factors were significantly different (p<0.01) in T1D cases vs controls; of these 11 were > 2-fold higher in T1D cases: Interleukin (IL)1β, IL6, IL10, IL13, IL15, IL23; Soluble CD40 ligand (sCD40L), tumor necrosis factor (TNF)β, epidermal growth factor (EGF), transforming growth factor (TGF)α and thrombopoietin (TPO). Using principal component analyses, case and control samples separated into two distinct clusters with little overlap, suggesting that cytokine profiles can accurately classify cases and controls. This was supported by partial least squares analysis, which estimates classification accuracy to be above 95% with leave-one-out cross-validation. Partial least squares analysis also ranked the contribution of each cytokine to accurate classification, in which the top 10 cytokines/chemokines were chemokine C-C motif ligand (CCL)8, sCD40L, TNFβ, chemokine C-X-C motif (CXCL)1, CXCL10, IL1β, IL10, IL13, TNF-related apoptosis-inducing ligand (TRAIL) and EGF. Conclusions: Our findings demonstrate that cytokine expression profiles may be useful biomarkers for classifying patients with T1D and indicate involvement of multiple immunological pathways in progression to T1D. Validation of these results in other T1D populations is needed.
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Cloning, Overexpression And Molecular Characterization Of Four Superantigens: Production Of Protein Therapeutics For Cancer Treatment
المؤلفون: Sara Bashraheel, Haya Al Sulaiti, Alanod Alqahtani, Morana Jaganijac, Wijnand Helfrich and Sayed K GodaBackground 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.
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Metalloproteinase-dependent And -independent Processes Contribute To Inhibition Of Breast Cancer Cell Migration, Angiogenesis And Liver Metastasis By A Disintegrin And Metalloproteinase With Thrombospondin Motifs-15.
Background & objectives. The ADAMTS proteinases are a family of secreted, matrix-associated enzymes that have diverse roles in the regulation of tissue organization and vascular homeostasis. Several of the 19 human family members have been identified as having either tumor promoting or suppressing roles. We previously demonstrated that decreased ADAMTS15 expression correlated with a worse clinical outcome in mammary carcinoma. Methods. We have explored the effects of A Disintegrin and Metalloproteinase with Thrombospondin motifs-15 (ADAMTS-15) on the behavior of MDA-MB-231 and MCF 7 breast cancer cells by stable expression of either a wild type (wt) or metalloproteinase-inactive (E362A) protein. Results. No effects on mammary cancer cell proliferation or apoptosis were observed for either form of ADAMTS-15. However, both forms reduced cell migration on fibronectin or laminin matrices, though motility on a Type I collagen matrix was unimpaired. Knockdown of syndecan-4 attenuated the inhibitory effects of ADAMTS-15 on cell migration. In contrast to its effects on cell migration, wt ADAMTS-15 but not the E362A inactive mutant inhibited endothelial tubulogenesis in 3D collagen gels and angiogenesis in the aortic ring assay. In experimental metastasis assays in nude mice, MDA-MB-231 cells expressing either form of ADAMTS-15 showed reduced spread to the liver, though lung colonization was enhanced for cells expressing wt ADAMTS-15. Conclusiona. These studies indicate that extracellular ADAMTS-15 has multiple actions on tumor pathophysiology. Via modulation of cell-ECM interactions, which likely involve syndecan-4, it attenuates mammary cancer cell migration independent of its metalloproteinase activity; however, its antiangiogenic action requires catalytic functionality, and its effects on metastasis in vivo are tissue niche-dependent.
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A Qualitative Exploration Of Facilitators And Barriers To Interprofessional Practice In Healthcare In Qatar
المؤلفون: Michael K. Corman and Jason HickeyBackground: 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.
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Specification Of Va And Vd Usage By Tcra/Tcrd Locus V Gene Segment Promoters
المؤلفون: Abbas Hawwari, Abani Naik and Michael KrangelThe Tcra/Tcrd locus undergoes V-Dδ-Jδ rearrangement in CD4-CD8- thymocytes to form the TCRδ chain of the γδ TCR and V-Jα rearrangement in CD4+CD8+ thymocytes to form the TCRα chain of the αβ TCR. Most V segments in the locus participate in V-Jα rearrangement, but only a small and partially overlapping subset participates in V-Dδ-Jδ rearrangement. What determines any particular Tcra/Tcrd locus V gene segment as a Vδ, a Vα, or both is currently unknown. We tested the hypothesis that V segment usage is specified by V segment promoter-dependent chromatin accessibility in developing thymocytes. TRAV15/DV6 family V gene segments contribute to both the Tcrd and Tcra repertoires, whereas TRAV12 family V gene segments contribute almost exclusively to the Tcra repertoire. To understand whether the TRAV15/DV6 promoter region specifies TRAV15/DV6 as a Vδ, we used gene targeting to replace the promoter region of a TRAV12 family member with one from a TRAV15/DV6 family member. The TRAV15/DV6 promoter region conferred increased germline transcription and histone modifications to TRAV12 in DN thymocytes and caused a substantial increase in usage of TRAV12 in Tcrd recombination events. Our results demonstrate that usage of TRAV15/DV6 family V gene segments for Tcrd recombination in DN thymocytes is regulated at least in part by intrinsic features of TRAV15/DV6 promoters, and argue that Tcra/Tcrd locus Vδ gene segments are defined by their local chromatin accessibility in CD4-CD8- thymocytes.
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RISK FACTORS, LIFESTYLE AND HEALTH HABITS OF YOUNG ADULTS IN QATAR
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.
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Inheritance Of Methylation In The Qatari Population
المؤلفون: Shaza Zaghlool, Mashael Al-shafai, Wadha Al Muftah, Pankaj Kumar and Karsten SuhreBackground & Objectives: The environment surrounding an organism can have a direct impact on methylation and trans-generational effects in both mammals [1] and plants [2, 3]. Some epigenetic changes are heritable or somehow passed on to future generations without altering the organism's underlying DNA sequence[4]. The concept of DNA methylation heritability in mammals is emerging, although the mechanism through which environmental impact is epigenetically transmitted is still not clearly understood. We search for methylation sites that exist in three states (trimodal) on a population level in a Qatari population for the first time. Using this subset of sites, we can test for a pattern of Mendelian inheritance of methylation, specifically by looking at trios. These patterns are then compared to randomized individuals where the trio structure was disrupted to conclude on the effect of family structure towards DNA methylation heritability. Methods: We used a Qatari cohort consisting of families. Some of the families contained trios (father, mother, and offspring). We obtained both methylation data (Illumina 450K array) [5] and whole genome sequencing data (Illumina Hiseq2500 platform) for the samples. We tested the 450K methylation sites for trimodality and filtered those satisfying the definition. The trimodal sites were then tested for Mendelian agreements and violations only in the trios. This was compared to randomized samples having no parent-offspring relationship as an enrichment study. Finally, we checked the trimodal sites for the likely possibility of being explained by an underlying genetic association. Results: We found less than 1% of the methylation sites across the dataset to meet the predefined trimodal criterion. We report that non-related individuals have a higher average of trimodal sites showing Mendelian violation in comparison with actual trios. Moreover, the average of trimodal sites exhibiting Mendelian agreement in trios is significantly higher in comparison with randomized samples. We also studied the effect of cis-acting SNPs on explaining the observed patterns of methylation heritability. Although in the majority of the sites, the pattern is elucidated by SNPs, there still a minute number of sites where the methylation is not explained by SNPs. These findings are replicated and validated in a bigger cohort. Conclusions: The great majority of methylation sites that follow a Mendelian inheritance pattern in trios are driven by an underlying SNP association. However, a minor percent of the trimodal sites we studied did not have a cis-acting SNP association although this does not rule out the effect of trans-acting SNP associations. We can speculate on other factors that could be causative of the trimodal pattern such as correlations between neighboring CpGs, SNPs otherwise in the genome, or even genome-wide probe-SNP eQTL associations.
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Influence Of Specific Hsc70 Domains On Fibril Formation Of Human Amylin Involved In Type 2 Diabetes: Importance Of The C-terminal Lid
المؤلفون: Ali ChaariProtein 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.
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A Prospective Metabonomics Analysis Reveals New Pathways Involved In T2D Development
المؤلفون: Abdelilah Arredouani, Loic Yengo, Beverley Balkau, Mario Falchi and Philippe FroguelBackground & Objectives: Characterization of the metabolic disruptions that precede the onset of T2D is critical for the screening of high risk individuals and hence the implementation of effective early interventions to prevent/delay the onset of T2D and its complications. Currently available predictors fail to grasp the complex etiology of T2D. Metabolomics profiling opened new horizons for biomarkers discovery; and associations of metabolomics and genotyping studies offers new prospects for the understanding of the physiology of complex traits like T2D. The aim of this study was the screening for novel predictors for T2D and the identification of associations between metabolic SNPs and metabolites as functional intermediate phenotypes. Methods: We used plasma collected from 1214 subjects from a 9 years follow up cohort for Non-targeted metabolomics. For genotyping we used a metabochip that assays nearly 200,000 SNPs of interest for metabolic disorders. Results, and Conclusions: We identified 491 named metabolites that are involved in 65 metabolic pathways. With a false discovery rate set at 5% (q<0.05) and after adjusting for BMI, sex and age, 109 metabolites were found to significantly associate with incident T2D (p<0.05). After adjusting for fasting glucose, 23 out of the 109 metabolites remained strongly associated with incident T2D (p from 3.12x 10-3 to 1.7x10-6). Of these, 1-palmitoylglycerol, 1,5-anhydroglucitol, 1-oleoylglycerol, mannose, alpha-ketobutyrate and gamma-glutamylphenylalanine showed the strongest associations ( p from 1.310-5 to 1.7x10-6). When using the metabolite ratios between year 9 and baseline after adjustment for fasting glucose, the metabolite 1,5-anhydroglucitol showed the strongest association with incident T2D (p=1.2x 10-9). Moreover, our analysis revealed a strong association of the metabolite mannose with the rs1260326 in the GCKR locus (p=8.8 10-40). The GCKR gene is considered a susceptibility gene candidate for T2D. We found associations of other metabolites with 8 different SNPs but none of them with known T2D loci.
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Induction Of Hydroxyurea-Mediated Altered Gene Expression-Like Pattern In Sickle Cell Anaemia Erythroid Cells From Qatari Populations.
المؤلفون: Queenie Fernandes, Mohamed Yassin and Nader Al-dewikBackground: 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.
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In-vitro Assembly Of A Sickle Cell Haemoglobin Intermediate To Mimic Wild-type Characteristics
المؤلفون: Queenie Fernandes and Daliya GeorgeBackground: Sickle Cell Anaemia (SCA) is a genetically-inherited class of haemoglobinopathies arising due to a point mutation in the nucleotides coding for the sixth amino-acid of the β-haemoglobin. During sickle cell haemoglobin (HbS) synthesis the amino acid "valine" gets incorporated in the growing protein chain at the site of "glutamic acid". At the final stages of protein folding to form the quaternary structure of HbS, the overall hydrophilic character of haemoglobin is affected. This instability causes obstruction in protein-protein interactions causing the HbS molecules to polymerise into long chains within the Red Blood cells (RBC). The formation of these long chain polymers of mutated HbS causes the distorted conformation of RBC turning it into sickled shape structures. The sickling of erythrocytes is the main cause of microcirculatory obstruction leading to painful vaso-occlusive crisis. Rationale: This study aims to target the improper folding of the HbS protein causing the overall imbalance in the otherwise neutral character of haemoglobin. This can be executed through partially unfolding the protein to expose the mutated base to the hydrophobic core. This will allow the residue to take an appropriate orientation preventing the polymerization of HbS and subsequent distortion of erythrocytes. Methods: Further to a preliminary study on HbA molecules, partial unfolding and refolding experiments were performed to identify and ensure that the quaternary structure of haemoglobin could be retained. The appropriate conformations of the modified HbS molecules were monitored through Circular Dichroism (CD) Spectrophotometry. Protein unfolding was carried out in the presence of Dimethyl Sulfoxide (DMSO); a mild denaturant. Unfolding was suspended at variable intervals of time by the addition of chloroform. Solubility tests were performed to look for aggregation due to polymerization. Results: It was previously identified that a change in structural conformation of HbS molecules was brought about by allowing unfolding for 36 to 48 hours. However the newly adopted conformation was not found to be analogous to that of the wild-type haemoglobin (HbA). The molecular stability was found to mimic wild-type characteristics. It was identified that the modified protein took relatively the same time through the C18 chromatographic column as compared to chromatographic tests performed on HbA molecules. Ideally HbA molecules are completely soluble in buffer (pH 7). The modified proteins were found to be insoluble in potassium phosphate buffer at room temperature, however there was no formation of aggregates which indicates that no polymerisation had taken place. Conclusion: Through this study it is evident that controlled unfolding of HbS to expose the mutated residue to the hydrophobic core of the protein is able to restore lost stability. However, the appropriate refolding of the protein is also important in order to restore the functionality of haemoglobin as its quaternary structure is a requisite for oxygen binding and transport. Thus, although unfolding of the protein may be successful in giving HbS a stable configuration, more refined protocols for arresting unfolding or supporting appropriate refolding is essential in order to ensure that the structural conformation of the protein is retained.
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Referral Pattern And 3D Visualization Of Microstructure Of Carotid Artery Plaque, Using Laser-Scanning Confocal-Microscopy (LSCM)
المؤلفون: Ahmed Khattab, Leopold Streletz, David Wertheim, Ibtisam Ali and Damian JenkinsonBackground: 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.
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An E-health Based Integrated Management Program Advancing Community Treatment Of Atrial Fibrillation (IMPACT-AF)
المؤلفون: Syed Sibte Raza Abidi, Jafna Cox, Samina Abidi, Ashraf Abusharekh and Joanna Nemis-whiteAtrial fibrillation (AF) is the most common abnormality of cardiac rhythm. There is growing evidence advocating an integrated multi-disciplinary approach to delivery of AF care facilitates which has led to improved care and outcomes and reductions in hospitalizations. In this research program, we have implemented a patient-centered and community-focused AF management program—termed as IMPACT-AF—to provide specialized AF care at the primary care level. Our approach is to exploit state-of-the-art e-Health technologies to: (a) computerize Canadian clinical practice guidelines as a web-based clinical decision support accessible to primary care providers to deliver evidence-informed AF diagnostic and therapeutic interventions at the primary care level; (b) monitor the patient in a home-based setting and proactively generate alerts and reminders to family physicians in response to adverse trends in the patient condition; and (c) engage patients through m-Health (mobile health) tools to self-manage their condition and undergo behaviour modification. Two research themes have been pursued in the development of the IMPACT-AF program: (1) Translation of AF Clinical Guidelines to Achieve Evidence-Informed AF Management: This theme involved three main tasks: (i) Computerization of the AF guidelines using ontology based knowledge models; (ii) Localization of the computerized AF guidelines to implement the local clinical pathways; and (iii) Operationalization of the computerized AF guidelines into practice through a computerized decision support system to provide evidence-informed patient care at the point-of-care. (2) Patient Engagement to Achieve Home-Based AF Management: This theme focused on the implementation of innovative AF self-management and behaviour modification interventions delivered to patients using mobile devices. We computerized theoretical behaviour modification model—i.e. Social Cognition Theory (SCT)—to generate personalized and mobile patient care plans. Using an AF (mobile) patient diary patients record their vitals and other AF related symptoms, and in turn receive educational messages, alerts, reminders and behaviour modification plans. The primary outcome of the IMPACT-AF project is reduction in AF related CV hospitalization. The secondary outcome is at two levels: (i) Process of Care: Reduction in specialist consultation, echo, catheter ablations; and, (ii) Economic: Reduction in health care costs and utilization for AF services. The clinical decision support systems developed by the IMPACT-AF project is deployed across the province of Nova Sctioa, engaging 200 primary care providers to deliver evidence informed AF management at the primary care level. We are gearing for a cluster randomized study involving 4000 patients across the province--the study will permit measurement and comparison of the clinical decision support system pre- and post-intervention, and across cases. The overall result will be cost-efficient improvement of care and outcomes for AF patients. In summary, the IMPACT-AF solution offers an innovative strategy to address two critical knowledge gaps —i.e. primary care physicians are often ill-equipped to provide evidence-based care for chronic conditions, and likewise patients are underprepared to apply the 'right' self-management and behaviour modification strategies to achieve meaningful outcomes in response to a longitudinal care plan. IMPACT-AF exploits state-of-the-art e-health technologies to develop an integrated and mobile clinical decision support systems to provide evidence-based AF care.
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A Guideline Compliant Clinical Decision Support System In Mobile And Smart Environments For Diagnosing Medical Conditions
المؤلفون: Patrice Roy, Newres Al Haider, William Van Woensel, Ahmad Marwan Ahmad and Syed Sr AbidiBackground & 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.
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Risk Of Foodborne Pathogens Associated With Retail Products In Qatar
المؤلفون: Kenlyn Peters, Darine Dimass, Ahmed Salem, Yu-chen Chang, Ali Sultan and Hussni Omar MohammedBackground and objectives-Foodborne illness has been determined to be one of the major limitations to the advancement of world health and with the ease of travel around the world and the increase in trade of food and animal products, the risk has been exacerbated in recent years. It has been proclaimed as one of the high priority issues in the Qatar National Food Security Master Plan. Different estimates of burden of disease consistently indicate a high cost per episode, irrespective of the country where the study is conducted. Bacterial pathogens among the leading causes of foodborne illness are E. coli, Salmonella, Campylobacter and Listeria. In an effort to understand the risk of these foodborne pathogens in the retail industry in Qatar, this study investigated the presence of these pathogens among retail products. Methods—Swabs collected from retail items were screened for the presence of different serotypes of these four foodborne pathogens using a combination of bacterial enrichment and real time polymerase chain reaction detection. Most of the samples were pre-cooked and collected from big retail stores in Doha. Specific tests were for E. coli O157:H7, the E. coli genes stx and eae, which are linked to Shiga toxin-producing E. coli (STEC), E. coli O26, O111, O121, O45, O103 and O145, Salmonella, Listeria, and Campylobacter jejuni, coli and lari. Results—E. coli O157:H7 was detected among the samples at a rate of 4.2%, E. coli genes stx (26.6%) and eae (26.3%), which are linked to Shiga toxin-producing E. coli (STEC) (16.6%), E. coli O26(5.9%), O111(3.5%), O121(1.4%), O45(20.1%), O103(2.1%) and O145(2.1%). These serogroups of E. coli non-O157 are known to be food adulterants and have zero tolerance in the US. In addition, Salmonella spp. were detected in 13.5% of the samples while Listeria monocytogenes were detected in 5.2% of the samples. Conclusions—Although the prevalence for many is low, the higher prevalence of STEC genes and STEC serotype O45 is cause for concern. Shiga toxin producing serotypes are becoming nearly as much of a concern as the more commonly known O157:H7. This demonstrates the importance of properly cooking meat products. Being exposed to foodborne pathogens can increase the risk of chronic gastroenteritis sequelae, including Inflammatory Bowel Disease.
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Cloning In E Coli, Overexpression And Molecular Characterization Of Novel Glucarpidase: Enzyme Involved In ADEPT For Cancer Treatment
المؤلفون: Alanod Alqahtani, Afrah Al-yafei, Mathew Groves, Alex Domling, Aishah Latiff and Sayed K GodaBack 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.
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Thyroid Function And Depression In Pregnant Women In Qatar
المؤلفون: Madeeha Nasir, Sundus Mari, Mariam Abdulmalik, Javaid Sheikh and Margaret AltemusContext There is an increased demand on the thyroid gland during pregnancy that results in lower levels of free thyroid hormone as pregnancy progresses. Thyroid peroxidase (TPO) antibodies, which are the most common cause of hypothyroidism and sub-clinical hypothyroidism, are found in 8-20% of women in the reproductive age group. It is unknown whether women with anti-thyroid antibodies are more prone to develop hypothyroidism during pregnancy. Hypothyroidism and sub-clinical hypothyroidism are associated with depression in men and non-pregnant women, but there has been little study of the degree to which low thyroid hormone or anti-thyroid antibodies increase risk of depression during pregnancy. Objective To determine whether low free T4 and TPO antibodies are associated with the development of depression during pregnancy. Design 371 non-depressed women were recruited from prenatal clinics in Doha, Qatar. Women were assessed during gestational weeks 8-16 (visit 1), 24-26 (visit 2) and 34-36 (visit 3). We screened women for depression using the Edinburgh Postnatal Depression Scale (EPDS). Full psychiatric diagnoses were established using the Mini International Neuropsychiatric Interview (MINI) at visit 1 for all subjects, and then repeated at later visits if the EPDS score was greater than 9. TPO antibodies were measured once at visit 1 and free T4 and TSH were measured at visit 1 and visit 3. Results 14% of subjects had TPO antibodies in the first trimester. The proportion of subjects who screened positive for depression (EPDS >11) was 16% at visit 1, 11% at visit 2 and 3% at visit 3. The proportion of subjects with major depression was 3.2% at visit 1, 3.6% at visit 2 and 0% at visit 3. In the 1st trimester, there was no relationship between any of the thyroid parameters and EPDS score, but in the 3rd trimester women with lower free T4 levels had higher EPDS scores (p<0.03). TPO antibody status was not related to EPDS score in any trimester. Free T4 levels dropped significantly from visit 1 to visit 3 (p<0.001) however, subjects with TPO antibodies did not show this decrease (p<0.01). Free T4 levels were lower in women that were TPO positive at visit 1, however, at visit 3 there was no difference in free T4 levels in women with TPO antibodies. Conclusion Anti-thyroid antibodies are common among pregnant women in Qatar and are associated with lower free T4 in 1st trimester but not later in pregnancy. In addition, TPO antibody status is not a predictor of increased risk of depression during pregnancy. However, free T4 does drop significantly from 1st to 3rd trimester and lower free T4 levels in the 3rd trimester are associated with higher EPDS depression scores. Women who develop depression during pregnancy should be screened for hypothyroidism. Thyroid hormone replacement during pregnancy is known to improve birth outcomes, but further work is needed to determine whether treatment of hypothyroidism during pregnancy will improve depressed mood or prevent postpartum depression.
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An Evaluation Of The Clinical Potential Of Ngs In Hcm
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.
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An Efficient Non-invasive Sample Collection For Various Population Segments
المؤلفون: Bassam El-fahmawiDeciphering genomes/proteomes at a personal level will not only help in identifying hereditary predisposing factors but also contribute to designing drugs with higher efficacies at the personal level. However, to arrive at this level of understanding we need to collect data in communities locally and globally. One of the major problems we face to achieve such objectives is volunteer recruitment for sample collection. Non-invasive sample collection represents the best option over blood in many ways: 1- Better volunteer compliance 2- No cold chain involvement for transport and storage 3- Self -collection and does not require expertise On the other hand, current non-invasive collection methods suffer from low DNA yields, quality and usability for the different segments of the population. Our innovative contribution with the iSWAB is focused towards simplifying non-invasive collection and thus increasing volunteer compliance while achieving high quality and yields of gDNA sufficient for several assays. In addition, the iSWAB design results in reduced sample processing time and cost in the lab, therefore allowing more samples to be processed per assay contributing to increased cost efficiencies for downstream high content genomic analysis.
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Gastric Sleeve Surgery Shifts Immunophenotyping Polarization From M1 To M2 With A Shift From Pro-inflammatory To Anti-inflammatory Pattern In Morbid Obese Subjects.
المؤلفون: Nasser Rizk, Moataz Bashah and Amina FadelBackground 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.
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"novel Genetic Variants May Modify The Clinical Outcome Of The Phospholamban L39x Mutation In Cardiomyopathy Patients"
Background Dilated Cardiomyopathy (DCM) is a leading cause for heart failure characterized by an enlarged ventricular cavity causing systolic dysfunction. Gene mutations are estimated to be the cause in approximately 30-50% of cases, while modifier genes are thought to influence the clinical outcome. Objectives Using a global and unbiased approach: Next Generation Sequencing (NGS), for one of the largest reported cardiac gene panels, QCRC aims at discovering novel gene variants implicated in DCM pathogenesis and/or progression. Method Through the QCRC Doha-centered intercontinental DCM patient cohort, we screened 38 DCM cases, confirmed by echocardiography, who did not have a history of alcoholism or coronary angiography findings. Patients were recruited following genetic counseling and informed consent, according to institutionally approved ethics protocols. High quality DNA was extracted from peripheral blood, 170 DCM known and candidate genes (=1.6Mbases) were sequenced using the HiSeq Illumina technology, and extensive bioinformatical analysis was pursued to depict genetic variations. Results We herein report an unusual case of a male DCM patient (LVEDD: 70mm; LVESD: 59mm; LVEF: 25%), diagnosed at age 40years, who developed sustained ventricular tachycardia (VT), was implanted an AICD at age 56years, and died age 60years. By NGS we determined that he was heterozygous for a known pathogenic nonsense mutation (c.116T>G) in the phospholamban (PLN) gene, which leads to a premature stop codon (L39X). The phenotype of L39X mutation carriers in the PLN gene has been shown to vary considerably, ranging from severe DCM, to rare reports of hypertrophic CM or normal cardiac function. However, this is the first occasion of L39X DCM patient presenting with sustained VT. Three additional, novel variants, out of the 163 variants detected in this patient, were of particular interest: i) a frameshift mutation (c.1495_1496insAGAC) in the C-terminus of CACNB2 (the beta subunit of the voltage-dependent calcium channel Ca(v)1.2). Mutations in this gene/protein region have been previously associated with Brugada syndrome with shorter than normal QT or early depolarization syndrome. ii) a non-synonymous SNP (c.9217C>T; p.L3073F) in laminin 2 (LAMA2), predicted to have a deleterious (SIFT) - possibly damaging (Polyphen2) effect. LAMA2 is a major component of striated muscle cytoarchitecture, and has been proposed, in a rare occasion, to relate to DCM with ventricular arrhythmias. iii) a non-synonymous SNP (c.6082A>G; p.T2028A) in the Alstrom Syndrome 1 (ALMS1) gene, predicted to have a deleterious (SIFT) effect. ALMS1 is implicated in the development of DCM, with rare reports of cardiac arrhythmias. Conclusion The co-presence of genetic variations in genes such as CACNB2, LAMA2 or ALMS1 may have an important modifier effect to the final clinical outcome towards DCM combined with arrhythmias.
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Personalized Medicine And Genomic Wide Association Study Based On Innovative Big Data Analytic And Data Mining Paradigm
المؤلفون: Emad A ElsebakhiPersonalized 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.
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Remote Slow Wave Sleep Monitoring Using A Single Electro-oculograph Channel Based System For Use With Insomnia
المؤلفون: Sana Tmar and Beena AhmedBackground and objective: Sleep disorders, such as insomnia can seriously affect an individual's performance and even lead to psychological problems. Diagnosis and treatment of sleep disorders require the collection of subjective and objective measures of sleep structure. Subjective measures are currently collected during face-to-face clinical consultations with a medical practitioner. Objective measures, obtained from polysomnographic (PSG) electrodes on the patient's body, are used to quantify sleep quality, specifically slow wave sleep (SWS) periods. In this work, we present an unobtrusive and ambulatory insomnia monitoring system which uses a single electro-oculograph (EOG) channel instead of a full PSG to identify SWS periods together with a mobile health application to collect subjective measures from patients. Method: Fig. 1 presents the architecture of our proposed system. It consists of three main components. The EOG sensing device records one EOG channel and wireless transmits it to the coordinator. The Coordinator (mobile phone) collects data from the EOG device and transmits it back to the clinical back-end. It also includes a mobile sleep diary application implemented on the Android OS to collect the subjective sleep assessment data from patients. The recorded data is time and date stamped and can be used for historical data analysis. The Clinical back-end consists of a database to store the received data and a server with a SWS classifier to assess sleep quality. Our SWS classifier is based on a simple rule based algorithm using spectral features extracted from several bands (alpha band (8 - 12 Hz), beta band (18 - 30 Hz) and delta band (0.5 - 4 Hz) with adaptive thresholds. We used EOG overnight sleep recordings from nine healthy subjects (18-64 years) to validate our SWS detector. Results: Our developed sleep diary was compared to existing sleep diaries applications. It was found to allow efficient handling of data with an improved layout and interface that enhanced user experience. Information collected through the diary also provided the clinician with better access to the required subjective data for an accurate diagnosis of insomnia. Our developed SWS detection algorithm was run on a testing group with 5 subjects and a validation group of 4 subjects. Sensitivity, specificity, relative observed agreement and Cohen's kappa coefficient values were computed and used to compare the output of the algorithm to the sleep experts' analysis. The agreement of our SWS detection method for the validation data was 90.0%, sensitivity 90.5%, specificity 89.9% and kappa value 0.74. Conclusions: Current PSG systems with multiple electrodes are inconvenient and uncomfortable for the user, resulting in modified sleep activity different from their normal night of sleep. Our work has shown that it is possible to reduce the complexity of the insomnia monitoring experience and still collect the required subjective and objective information needed to assist in insomnia diagnosis. Acknowledgement: The work was supported by NPRP grant #[5-1327-2-568] from the Qatar National Research Fund which is a member of Qatar Foundation. The statements made herein are solely the responsibility of the authors.
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Design, Expression And Characterization Of A Single Chain Fragment Variable Anti-mcf-7 Antibody; A Humanized Antibody Derived From Monoclonal Antibody
المؤلفون: Ilham Omer MahgoubAbstract 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.
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Hyperglycemia Regulates Annexin A2-s100a10 Localization In Endothelial Cells
المؤلفون: Lara Bou Khzam, Katherine Hajjar and Nasrin MesaeliLara Bou Khzam1, Katherine Hajjar2, Nasrin Mesaeli1 1Weill Cornell Medical College in Qatar, Education City, Doha, Qatar and 2Weill Cornell Medical College New York, USA. Diabetic retinopathy (DR) is a leading cause of legal blindness in working-age individuals resulting in disrupted vascular integrity and pathological retinal angiogenesis in both type I and type II diabetes. Annexin A2 is a regulator of endothelial morphogenesis and plays a role in the modulation of fibrin homeostasis and neoangiogenesis by regulating the generation of plasmin. Heterotetramerization of S100A10 (p11) with Annexin A2 and translocation of this complex to the cell surface is required to modulate the activation of plasmin. We hypothesize that the Annexin A2-p11 system regulates the development of retinal angiogenesis and may be a potential therapeutic target in diabetic retinopathy. Objective of current study is to determine the effect of hyperglycemia on Annexin A2 and p11 expression in endothelial cells. To address this objective we examined effect of high glucose on Annexin A2 and p11 expression in HUVEC cells. Our results illustrated no significant change in the total Annexin A2 protein or mRNA after exposure to different doses of glucose for different times. However, hyperglycemia increased p11 protein expression. To determine weather hyperglycemia affects cellular distribution of Annexin A2 and p11 we used cell surface biotinylation and immunocytochemcial staining (ICC) . Biotinylation studies illustrated an increased cell surface expression of both Annexin A2 and p11 under high-dose glucose conditions. ICC experiments also illustrated cell surface localization of Annexin A2 and p11 following high glucose exposure. Overall our data illustrates a role for high glucose in post-translational regulation of Annexin A2 and p11. Future studies are focused on examining the nature of the post-translational modification in Annexin A2 and p11. Acknowledgment: This work is supported by NPRP No: 6-736-3-187 by QNRF.
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Changing Nursing Practice
المؤلفون: Merry-jo LeversDescription: 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.
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A Novel Homozygous Lrp5 Splice-site Deletion Mutation Causes Syndromic Autosomal Recessive Familial Exudative Vitreoretinopathy.
المؤلفون: Vasiliki Chini, Yasser Al Sarraj, Michael Trese, Hatem El Shanti and Marios KambourisA consanguineous Saudi Arabian family with two female siblings affected by an autosomal recessive condition resembling Familial Exudative Vitreoretinopathy [FEVR], but also with short stature, bone fragility with thin and wasted appearance was studied by homozygosity mapping and positional candidate gene screening to identify the offending gene and mutation. The gene was mapped to three possible homozygous genomic regions [[2q, 4q, 11q], as the family structure did not allow identification of a single interval with a significant LOD score. Mutations in three genes (FZD4, TSPAN12, NDP and LRP5) have been associated with FEVR. The LRP5 gene localizes within the 11q13.2 homozygosity interval in this family rendering it the positional candidate of choice. Screening by Sanger sequencing identified a novel homozygous one-base splice-site deletion mutation c.3236+1 delG in exon 14. LRP5 is a low-density lipoprotein receptor (LDLR) a transmembrane protein that binds and internalizes ligands in the process of receptor-mediated endocytosis. The cDNA encodes a 1,615-amino acid protein containing conserved modules including a putative signal peptide, four epidermal growth factor (EGF) repeats with associated spacer domains, three LDLR repeats, a single transmembrane-spanning domain, and a cytoplasmic domain. The extracellular domain contains 6 potential N-linked glycosylation sites. LRP5 has a unique organization of EGF and LDLR repeats compared to other LDLR family members and in addition to FEVR, mutations in the gene have been associated with Hyperostosis corticalis OMIM 144750; Osteopetrosis, autosomal dominant 1 OMIM 607634; Osteoporosis-pseudoglioma syndrome OMIM 259770; Osteosclerosis OMIM144750; van Buchem disease, type 2 OMIM 607636; Bone mineral density variability 1 OMIM 601884; Osteoporosis OMIM 166710. Only missense mutations and splice site substitutions in LRP5 have been associated with autosomal dominant and recessive FEVR. This is the first report of an autosomal recessive LRP5 splice-site deletion mutation causing a syndromic form of FEVR.
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Title: The Use Of Low Intensity Pulsed Ultrasound (lipus) And Gingival Mesenchymal Stem Cells (gmscs) For The Treatment Of Severe Periodontal Defects In Dogs
المؤلفون: Khaled Ataf Abdel-ghaffar, Tarek El-bialy, Ali Saleem, Mamdouh Farid and Elham FawziAbstract: 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.
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Characterizing The Role Of Fikk Kinases In Toxoplasma Gondii Encystation
المؤلفون: Sini Skariah, Dana Mordue and Ali SultanToxoplasma gondii (Tg), causative agent of the disease toxoplasmosis in humans is an opportunistic pathogen belonging to the phylum Apicomplexa, which includes many medically important pathogens such as Plasmodium. Tg life cycle is characterized by two cysts stages: tissue cysts containing bradyzoites and oocysts. Taken together tissue cysts in contaminated food/water and oocysts shed into the environment are responsible for nearly 25% of the adult population worldwide being chronically infected with this parasite. Clinically, encystation by Tg presents the greatest challenge towards treatment of Tg as the cyst form is currently untreatable. Our aim is to identify genes and regulatory networks that regulate Tg oocyst/tissue cyst production. Isolation of such global regulators, which regulate encystation will be crucial for designing therapeutic strategies which effectively prevent cyst formation and block transmission of the disease by felines. Using a forward genetics approach, we have isolated a unique Tg mutant, named 11BE9, that under normal growing conditions (tachyzoite stage) in vitro dramatically expresses more than 100 genes, that are usually expressed in the oocyst/bradyzoite stages. Our hypothesis is that a global regulator(s) has been deregulated in this mutant, which is responsible for the over-expression of cyst-associated genes in the tachyzoite stage in the mutant. To identify this regulatory gene, we compared the transcriptome of this mutant to wild type Tg using microarrays, and Tg FIKK kinase gene was identified as a potential candidate. FIKK kinases are novel serine threonine kinases that share a conserved stretch of amino acids and are hypothesized as drug targets due to their unique expression in Apicomplexa. Our preliminary studies show that deletion of the FIKK gene in Tg leads to an impaired in vitro conversion of the tachyzoites to bradyzoites which is required for the establishment of the chronic cyst stage of the parasite during toxoplasmosis. Currently we are in the process of complementing the gene deletion clone to confirm the role of Tg FIKK kinase on cyst formation.
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