Qatar Foundation Annual Research Forum Volume 2013 Issue 1

ABSTRACT Background and Objective HIV and herpes simplex virus type-2 (HSV-2) are sexually transmitted infections, with HSV-2 being nearly 10 times as infectious as HIV. HSV-2 prevalence has been established as an objective biomarker and a proxy measure of sexual risk behavior. HIV epidemics appear to be emerging in the Middle East and North Africa (MENA) region, but the time course and future size of these epidemics are still unknown. We assessed HIV epidemic potential across regions using the association between HSV-2 and HIV prevalence, and based on a global systematic review of HIV and HSV-2 prevalence and meta-analyses. Methods Sources of data were PubMed and Embase databases, and country-level reports. Data on the prevalence of HIV (PHIV) and of HSV-2 (PHSV2) in the same population were extracted from eligible studies. Our analysis included all population groups except adolescents and injecting drug users, where the dominant mode of HIV transmission was not sexual. Studies with sample sizes of less than 50 were excluded. Odds ratios were calculated as (PHSV2/ (1- PHSV2)) / (PHIV/ (1- PHIV)), and relative risks as (PHSV2/ PHIV). Meta-analyses were conducted incorporating the inverse variance method and using a random-effects model to pool summary estimates of odds ratios and relative risks in different geographical regions. Results The meta-analysis included 662 study measures with a total population of 590,220 individuals from 67 countries. The pooled odds ratio of HSV-2 prevalence relative to HIV prevalence was low in Sub-Saharan Africa, Asia, and North America/West Europe, 6.0 (95% CI 5.5-6.5), 10.4 (95% CI 9.2- 11.8), and 13.0 (95% CI 9.9- 17.8), respectively. We found high odds ratios in East Europe 73.6 (95% CI 32.8-165.5), MENA 38.0 (95% CI 15.2-95.0), and Latin America 30.4 (95% CI 22.4-41.4). The pooled estimates of relative risks followed the same pattern as the odds ratios across all regions. Discussion The small odds ratios in sub-Saharan Africa, Asia, and North America/West Europe suggest established HIV epidemics that have likely reached saturation. Meanwhile, the large odds ratios in MENA, East Europe, and Latin America, suggest emerging HIV epidemics that have not yet reached their epidemic potential. These results corroborate recent data about the trend of HIV epidemics in these regions. The findings are of concern for MENA as they suggest that the HIV epidemics are likely to grow within this decade and the next one. HIV response in MENA needs to be expanded through bio-behavioral surveillance, voluntary counseling, testing, and treatment services, and focus on HIV prevention.

Health-related quality of life (HRQoL) measures are important indicators of perceived physical and mental health and are affected by changes in health status, demographic characteristics, culture, and ethnicity. Although recent studies have reported on HRQoL in midlife women in the US, most notably the Study of Women's Health Across the Nation (SWAN), there has been little research conducted on quality of life issues experienced by midlife women in the Middle East. Qatar has a high prevalence of obesity, a known risk factor for many chronic diseases, including osteoarthritis (OA) and rheumatoid arthritis (RA), which adversely influences HRQoL. This study has the following goals: To compare HRQoL among midlife Arab women living in Qatar to that of women in the US, (2) to report the prevalence of osteoarthritis (OA), rheumatoid arthritis (RA), and symptoms of aches and stiffness in joints, (3) to examine the impact of OA and RA on HRQoL, and (4) to compare arthritis prevalence and HRQoL of Qatari women to that of non-Qatari Arab women living in Qatar. This is a cross-sectional study of 841 women aged 40-60 recruited from nine primary health centers. Height and weight were assessed by physical examination and body mass index (BMI) was calculated as weight (in kilograms) divided by the square of height (in meters). Face-to-face interviews included demographic questions as well as questions about whether women suffered from any of a list of medical conditions, including OA and RA. The SF-36 was used to assess HRQoL and eight subscales were computed. A higher score designates a more favorable health status. Overall, women in Qatar had significantly lower mean scores than women in the SWAN for bodily pain (53.0 vs 68.9, p=0.0001) and for vitality (49.6 vs. 54.8, p=0.0001). BMI = 35 was found in 41.4% of women. OA and RA were reported by 4.8% and 4.3% of women, respectively, yet 71.6% reported being bothered by aches or stiffness in joints within the past two weeks. Half of women reported that these symptoms were either quite bothersome or intense. Women who reported having OA or RA had significantly lower scores on physical function (p<0.0001 and p=0.0002, respectively). Those with RA also had significantly reduced functioning for role physical (p=0.001). There was a trend toward lower scores among women with RA on bodily pain, vitality, and social functioning and among women with OA on bodily pain compared to women not reporting arthritis. Qatari nationals had a significantly higher prevalence of OA compared to non-Qatari women (7.4% among Qatari nationals vs. 2.6% among non-Qatari women, p=0.001). Women of Qatari nationality reported lower mean scores on physical function than non-Qatari women (73.3 vs. 80.0, p<0.0001). Midlife women in Qatar reported more bodily pain and reduced vitality than women in the SWAN, which may be the result of joint pain. The high prevalence of obesity may be predisposing women in Qatar to arthritis, which appears to be underdiagnosed given the high prevalence and intensity of joint aches and stiffness. National surveillance of the population is warranted.

All muscles involve the interaction between two sets of filamentous proteins, actin and myosin, that leads to muscle contraction and force production mediated by the hydrolysis of ATP (Adenosine triphosphate). Actin filament structure is understood to high resolution, but myosin filament structure is much less well defined. The myosin filaments are formed from complicated arrangements of myosin molecules and accessory proteins. Myosin molecules are hexameric polypeptide chains, each consisting of two myosin heavy chains (MHC) and four myosin light chains (MLC). Accessory proteins are the myosin binding protein C (MyBP-C) and Titin. We have previously resolved the three-dimensional (3D) structure of myosin filaments in normal human heart muscles (AL-Khayat et al., Proc. Natl. Acad. Sci., USA, 110, 318-323). Mutations in cardiac muscle myosin (MHC or MLC) and its associated proteins (MyBP-C or Titin) are known to be associated with a number of myopathies (e.g. familial hypertrophic cardiomyopathy and dilated cardiomyopathy) which change the proteins involved in producing and regulating heart muscle contraction. In order to understand the effect of myosin-associated heart disease, it is important to understand the 3D structure of myosin filaments in both the normal as well as in the diseased human heart muscles. The aim of this project is to study and compare the arrangement of the myosin molecules and the accessory proteins in the human heart muscle myosin filaments and how these arrangements change in diseased human heart muscles, suffering from either hypertrophic or dilated cardiomyopathies. A laboratory method to isolate myosin filaments from normal undiseased human cardiac muscle that preserves the highly ordered pseudo-helical structure of the myosin filaments has already been developed. This led, for the first time, to the detailed analysis of the 3D structure of myosin filaments from normal human heart muscles by utilizing the experimental technique of transmission electron microscopy (EM) as well as the computational single particle image analysis, 3D reconstruction and structural interpretation. Knowledge of this 3D structure serves as the starting point from which myosin filaments isolated from human cardiomyopathic samples, with known mutations in either myosin or its associated proteins, will be studied later in detail. The 3D structure of mutated myosin filaments will be resolved by a state of the art Electron Microscopy Facility at QCRC which is now underway. This will have both the highest spatial and time resolution for collecting EM images. A laboratory method will also be developed to isolate myosin filaments from human cardiomyopathic samples, which will be examined by EM and single particle image analysis so that to determine the overall 3D structure of myosin filaments in diseased heart muscles. By direct comparison to the known 3D structure of myosin filaments from normal undiseased human cardiac muscle, this would eventually permit the structural effects of known myosin filaments-associated mutations to be investigated in detail as well as to relate structure-to-function-to the overall disease process. Detailed understanding of the disease process would then allow us to design possible.

Background: EEG signal are widely used for detecting abnormalities such as seizures in newborn babies. EEG signals have non-stationary characteristic, therefore time frequency distributions (TFDs) are a preferred tool for their analysis. The classification performance of most time-frequency (t-f) methods is restricted by the resolution limitation of TFDs. Objective: The main objective of this research is to propose a new time-frequency pattern recognition technique that improves the performance of earlier methods by defining a new high resolution data adaptive time-frequency distribution. Methods: The key steps of the proposed t-f pattern recognition scheme are 1. Transformation of an EEG signal in the t-f domain by using a high resolution TFD. 2. Estimation of the instantaneous frequency (IF) and instantaneous amplitude (IA) of signal components. 3. Extraction of statistical features such as mean, variance, skewness and kurtosis from the estimated IF and IA. 4. Training of a support vector machines using extracted features. The accurate estimation of the IF and IA of signal components is a key step of the proposed t-f pattern recognition technique. Estimation of the IF and IA of signal components depends on the ability of a TFD to resolve closely spaced signal component. In order to overcome the resolution limitation of existing TFDs, we define a new high resolution data adaptive TFD that adapts the direction of its smoothing kernel at each point in the t-f plane based on the direction of energy concentration in the t-f plane. The proposed adaptive TFD out performs other standard methods of t-f analysis in terms of its resolution and instantaneous frequency estimation capabilities. Results: The proposed t-f pattern recognition methodology is applied to detect seizure activity in newborn EEG signals. The classification performance of widely used TFDs such as the extended modified B-distribution, compact support kernel, spectrogram, and proposed adaptive TFD is compared using the leave-one out cross-validation technique. The proposed TFD outperforms other TFDs as well as earlier methods of seizure detection by achieving the total accuracy of 97.5%. Conclusions: A new time-frequency pattern recognition technique for the classification of EEG signals is presented. Results indicate that the performance of the time frequency pattern recognition techniques is sensitive to the resolution performance of TFDs as high resolution TFDs have achieved better classification results.

Background. Ovarian cancer is the second leading cause of cancer-related death in women worldwide. Despite optimal cytoreduction and adequate adjuvant therapy, initial tumor response is often followed by relapse. Targeted therapies have been evaluated in ovarian cancer to overcome resistant disease. Among them anti-angiogenic therapies inhibit new blood vessel growth, induce endothelial cell apoptosis, and block the incorporation of haematopoietic and endothelial progenitor cells into new blood vessels. Despite in-vitro and in vivo successes anti-vascular therapy with bevacizumab targeting VEGF has limited efficacy in ovarian cancer. Anti-angiogenic treatment increases hypoxia, and might lead to tumor rebound and drug resistance. The precise molecular mechanisms underlying clinical resistance to anti-VEGF therapies are not well understood. But it became clear that the multiple changes in the stroma may determine the treatments outcome, Hypothesis. We hypothesized that abnormalities in the tumor endothelium may contribute to treatment resistance and produce and promote a residual microscopic disease and resistance to bevacizumab. Methods. We showed that Akt pathway is activated in vitro and in vivo in ovarian cancer endothelium. We used Akt-activated endothelial cells (E4+EC) that replicate tumor endothelium biology, and their control, HUVEC to investigate the anti-angiogenic activity of bevacizumab by angiogenesis and migration assays. We conducted XTT assay to examine the effect of bevacizumab on proliferation of VEGF producing human ovarian cancer cell lines. Expression of FGF-2, phospho-AKT was assessed by western blotting and flow cytometry. Finally, using a feeder-free matrigel and spheroid models of ovarian cancer we examined the effect of bevacizumab on residual disease. Results. Our study describes a comprehensive observational and functional investigation on the pivotal role played by the endothelium in the resistance to bevacizumab. We showed that the cross-talk between ovarian cancer cells and the endothelium activate PI3k/Akt. According to our findings, activated ECs expressing higher amount of VEGF-A tend to be less susceptible to the inhibitory effect of bevacizumab. Bevacizumab had no effect on the proliferation of Akt-activated EC, but significantly inhibited angiogenesis and delayed wound healing in HUVEC. We were able to show most primary ovarian cancer cells and ovarian cancer cells cultures secrete a large quantity of FGF-2 and showed that FGF-2 is able to revert the effect of bevacizumab on HUVEC. Our data suggest that an FGF-2/FGFR mediates a cross-talk between cancer and endothelium and is inviolved in an angicrine switch. We demonstrate the role of Akt-activated EC in supporting expansion and self-renewal of OCC in a residual disease context. Conclusion. We used the E4+ECs as a surrogate for tumor associated endothelium. We showed that an FGF-2/PI3K-AKT autocrine loop is required in ECs to perturb bevacizumab treatment. In summary, our study point out the role of an activated endothelium in the constitution of the residual disease and resistance to bevacizumab.

Both diabetes and cancer are prevalent diseases whose incidence is increasing worldwide and especially in countries that are undergoing rapid industrialization (i.e. Golf Countries). Lifestyle risk factors including diet, physical activity and obesity play a pivotal role in the etiology of both diseases. Epidemiological studies provide strong evidence that subjects with diabetes are at significantly higher risk of developing many forms of cancer and especially solid tumors. In addition to pancreatic and breast cancer, the incidence of colorectal cancer is increased in diabetes. A person with diabetes has a 38% higher risk of developing colon cancer compared to other people. Male diabetes patients were found to have a 20% higher risk of developing rectal cancer. While diabetes and especially type 2 diabetes and cancer share many risk factors, the biological links between the two diseases are poorly characterized. We have evidence that in human epithelial colorectal cancerous cells, either high glucose, insulin or their combination inactivates adenine monophosphate kinase (AMPK), induces the loss of function of the tumor suppressor gene, tuberous sclerosis complex 2, encoding tuberin, activates the mTOR/S6Kinase pathway and enhances the generation of mutagnic DNA, 8-oxodG. We also show that AMPK inactivation upregulates Nox1, which in turn inactivate tuberin and mTOR. These observations were associated with increased cellular proliferation, migration and fibronectin accumulation. Treatment of the cells with metformin, a potent AMPK activator or with rapamycin, an mTORC1 inhibitor, decreases the rate of cellular proliferation and migration and reverse the biochemical changes seen in cancerous cells treated with high glucose, insulin or their combination. In rodent models of diabetes, we find that loss of function of tuberin is associated with loss of function and mutations of OGG1 gene and accumulation of significant amounts of 8-oxodG and activation of the mTOR/S6Kinase pathway. These observations are paralleled by an increase in the levels of ROS production through an NADPH dependent mechanism. These observations indicate a critical role for AMPK, tuberin and mTOR in diabetes progression. These same observations were found in animal models that develop spontaneous colorectal cancer, APC mice. Activation of AMPK or blockage of mTORC1 pathways in the APC mice decreases ROS production, reverse OGG1 mutation and 8-oxo-dG accumulation in the colon and decrease tumor development. Our Findings may suggest that activation of AMPK, blockage of mTORC1 or inhibition of the NADPH oxidases pathways represent potential targets to reduce or to inhibit the onset and the progression of colorectal cancer in diabetes.

Prostate cancer (PC) is the second leading cause of cancer related death in men in several countries and is the most common cancer in men in Qatar according to recent statistics of the years 2010 and 2011. Understanding the molecular mechanisms underlying the development of PC progression is critical for developing novel therapies. Neuroendocrine (NE) differentiation of PC cells is an oncogenic process that has been reported as a mechanism contributing to hormone refractory PC progression. NE cells are found in most prostate adenocarcinoma that are significantly enriched as disease progresses, and are extremely resistant to apoptosis caused by androgen ablation therapy, therapeutic drugs or radiation. However, the mechanism of resistance to apoptosis of these cells is not clearly understood resulting in major impediment in PC treatments. The current study aims to initially identify the optimal conditions to induce NE phenotype in PC cells in vitro and to identify the molecules or pathways responsible for resistance to apoptosis in NE cells of prostate adenocarcinoma. Since NE cells are also quiescent as cancer stem cells (CSC), we investigated the relation between NE cells and CSC. Using the expression of NE-specific markers such as Chromogranin A and Neuron specific Enolase (NSE) an optimal method for the differentiation of PC cells to NE cells was identified and used to generate a repertoire of NE-induced PC cells for genome-wide analysis. Our analysis identified differential regulation of pro-apoptotic and anti-apoptotic molecules in NE cells accounting for resistance to apoptosis. We also show that the NE repertoire is highly enriched in CSC as demonstrated by sphere formation method. The identification of key molecules involved in resistance to apoptosis is critical for the development of novel therapeutic drugs that can interfere with NE differentiation and thus PC progression.

Background: Egypt has by far the highest national-level hepatitis C virus (HCV) prevalence in the world, with more than 14% of the general population infected with the virus. The drivers of such epidemic are still not completely identified, with previous parenteral antischistosomal therapy (PAT) campaigns conducted throughout Egypt proposed as the major contributor of the high HCV prevalence. In an effort to clarify specific drivers of the HCV epidemic in Egypt, and to identify priority populations for HCV prevention interventions, we conducted a comprehensive statistical analysis and mapping of the spatial distribution of HCV infection across Egypt. Methods: We conducted statistical analysis of the world's largest cross-sectional study of HCV conducted by the Demographic and Health Survey in Egypt (EDHS) in 2008 including demographic, health, and HCV biomarker information for a sample of over 11,000 individuals aged 15-49 years. We also identified and compared spatial clusters with high numbers of HCV infections using Kulldorff spatial scan test. The test locates areas with higher numbers of HCV infections than expected under spatial randomness. For each identified cluster, a likelihood ratio test was computed. A P-value was determined through Monte Carlo simulations to evaluate the statistical significance of each cluster. Results: The analysis of the EDHS data indicated that only 9.1% of the population aged 15-59 was exposed to PAT; from this fraction, 28.6% are infected with HCV. These values suggest that PAT might be responsible for only about 8% of the total HCV infections in Egypt. Moreover, ever had a blood transfusion, ever had a dental treatment, and ever had a surgery significantly increased the odds of testing positive for HCV by 54%, 45%, and 28%, respectively. Our results also indicated substantial geographical clustering of HCV infections in Egypt. We identified settings with high HCV prevalence distributed in six geographical clusters located at the interface between the governorates of Beni Suef and Minya, Faiyum, Dakahlia, Kafr el-Sheikh, Monufia, and Minya. Conclusion: The small fraction of the population with previous exposure to PAT suggests that other modes of transmission fueled the HCV epidemic, and could be playing an essential role in the current spread of the HCV infection in Egypt. Our study provides direct evidence for strong geographic clustering of HCV infection in Egypt and locates priority geographic areas for spatially targeted interventions.

Abstract Objective: To compare the efficacy and safety of 5%, 3%, and 0.9% saline solution for treating acute bronchiolitis in the prehospital setting. Study design: This was a double-blind trial including consecutive infants aged <18 months treated in an urban urgent care setting. A total of 165 patients were randomized to receive nebulized 5%, 3%, or 0.9% (normal) saline with epinephrine every 4 hours. The primary efficacy outcome was bronchiolitis severity score improvement at 48 hours (c2 analysis). Scores and oxygen saturation immediately before and after each treatment were recorded to assess safety. Results: A total of 187 previously healthy infants (median age, 3.1 months) diagnosed with bronchiolitis were enrolled. Positivity for respiratory syncytial virus was similar in the 3 treatment groups (mean, 56%). At 48 hours, the mean severity score for the 5% saline group was 3.69 _ 1.09, and that for the 0.9% saline group was 4.12 _ 1.11 (P = .04; difference, 0.43, 95% confidence interval for the difference, 0.02-0.88). The mean severity score for the 3% saline group was intermediate at 4.00 _ 1.22. Revisit rates after discharge were similar in the 3 treatment groups. No adverse reactions or other safety concerns were identified. Conclusions: Nebulization with 5% hypertonic saline is safe, can be widely generalizable, and may be superior to current treatment for early outpatient treatment of bronchiolitis.

Introduction Colorectal cancer (CRC) is increasing in incidence in several Asian countries but screening programs are lacking. In Qatar, ,we embarked on a pilot study for Colo-Rectal Cancer (CRC) screening based at 3 primary health centers among subjects with average risk for colorectal cancer. Aim To devise a national screening program for the early detection of colonic cancer among persons at average risk for Colorectal cancer (CRC) ,based on the outcome of our pilot study Methods All subjects aged 40-74 years at average risk for CRC ie that is individuals without history of polyps or documented cancer or inflammatory bowel disease in the past and absent family history are included. They are subjected to screening by immunochemical fecal occult blood testing at the primary health centers. Subjects positive for stool occult blood undergo total colonoscopy at the tertiary care hospital. Cancers and polyps picked up are documented and subjected to appropriate investigations and therapy Results A total number of 1242 healthy subjects at average risk for CRC, between ages of 40-74 years underwent stool occult blood testing by immunochemical fecal occult test. 57 (4.5%)were found to be positive and referred for colonoscopy. Of the 57 eligible persons, invited for colonoscopy, 32(56.4%) underwent the procedure , of whom 7 had polyps and 5 patients had cancer or advanced adenoma. Conclusions Cancers and adenomatous polyps which are treatable and curable can be successfully picked up by primary care level screening programs. Relevance, feasibility and cost effectiveness of such screening programs in Middle East has to be confirmed by community based programmatic screening studies in our region