Qatar Foundation Annual Research Forum Volume 2012 Issue 1

Objectives: Human papillomavirus (HPV) is the most commonly known sexually transmitted agent. To date, few reports are available on the distribution of most prevalent and variant types of HPV in Arab women. Therefore, the aim of this study is to determine the age-specific distribution of HPV types among Arab women being subjected to routine pap smear tests in the State of Qatar. Methods: 1100 pap smears have been collected in ThinPrep vials (BD SurePath™) from Arab women seeking routine gynecological care at the Women's Hospital, Hamad Medical Corporation (HMC) in Doha, Qatar. All the samples were transported to the laboratory in an icebox container. Viral DNA from ThinPrep samples was extracted by QIAamp MinElute virus spin kit according to manufacturer's instructions and was screened for HPV DNA by real-time PCR using L1 HPV specific (GP5+/6+) primers. The type-specific distribution of the viruses was determined by HPV high and low risk typing RT-PCR kits (Sacace Biotechnology, Italy) and PCR-based sequencing. Real-time PCR amplification was carried out in ABI 7500 real-time PCR (Applied Biosystems). Results: Based on the collected data, HPV DNA was detected in 125 women (11.36%), and 15 different HPV genotypes were detected, comprising high-risk and low-risk genotypes. The prevalence of HPV infection was seen in 54.5% Qatari and 45.5% non-Qatari women. With regard to age, 33.5% of all HPVs were found in women 30-39 years of age, 24.8% in women 40-49 years of age, 15.8% in women 50-59 years of age, 5.5% in women over 60 years and 20.4% in women less than 30 years old. Implications and Impact: The study shows that the prevalence of HPV infection in Arab women in Qatar is among the highest in the Arab world compared with previous reports. However, more extensive population-based studies should be undertaken before implementing HPV vaccination.

Background and Objective: The State of Qatar has achieved maternal, neonatal and perinatal survival rates which are comparable to many high income countries, both from the West and East. Our study aims to analyze fetal and perinatal determinants of Qatar's neonatal mortality rate (NMR) during 2011. Methodology: A PEARL Study (Perinatal Neonatal Outcomes Research Study in the Arabian Gulf), a joint collaborative research project between Hamad Medical Corporation (HMC) Qatar and University of Gloucestershire United Kingdom, is Qatar's prospective national perinatal epidemiological Study funded by Qatar National Research Fund. The study is quantifying maternal, neonatal and perinatal mortality, morbidities and their correlates by establishing a national neonatal perinatal registry for Qatar called Q-Peri-Reg. Data on live births and neonatal mortality were collected from all public and private maternity facilities in Qatar during 2011. Data on fetal and perinatal determinants was ascertained from maternal obstetric and delivery room record on predesigned performas. Univariate and multivariate regression analysis was done using Epi Info and SPSS-20. Results: Qatar's NMR during 2011 was 4.9. The incidence of low birth weight in Qatar is 11% and the incidence of preterm deliveries 10.7%. 10% of the babies required delivery room resuscitation. The relative risk of neonatal mortality was higher and statistically significant with fetal growth (p<0.001), fetal weight at birth (p<0.001), fetal gestation at birth (p<0.001), APGAR score at 1 and 5 minute (p<0.001) and the need for delivery room resuscitation (p<0.001). The RR of neonatal mortality increased (Table 1) with decreasing birth weight (p<0.001) and gestational age (p<0.001). Conclusion: Further improvement in Qatar's neonatal mortality is possible by addressing the high incidence of low birth weight and preterm deliveries. Better maternal nutrition, improved antenatal care, birth spacing and best obstetric and neonatal practices at delivery are likely to be most helpful.

Background: The AGE reader is a non-invasive device that measures tissue accumulation of advanced glycation endproducts (AGEs) through skin auto-fluorescence (AF) and gives prediction of cardiovascular risk. For the risk prediction, the AGE reader uses a single reference curve, for both females and males, obtained from Caucasian subjects. Based on these reference curves, clinical cut-offs for a low, medium and high AF have been made. This study examines whether these reference values can be used for ethnic populations seen in Qatar. Furthermore, we assess whether gender and smoking affect skin AF in this particular population. Methods: Skin AF was measured in 200 Arabs, 99 South Asians and 35 Filipinos. Using multivariate linear regression analysis and adjusting for the covariates age and the presence of type 2 diabetes, we assessed whether ethnicity, smoking and gender were associated with skin AF. Results: The Arabs and the Filipinos had a significant higher skin AF then the South Asian population (0.272 (95% CI: 0.138, 0.406), p <0.001 and 0.354 (95% CI: 0.147- 0.561), p=0.001 respectively). This is equivalent to a horizontal shift of 14.6% and 19.0%, respectively. Also, skin AF was significantly higher in women compared to men (0.432 (95% CI: 0.307, 0.558), p <0.001). Smoking was positively associated with skin AF (0.21 (95% CI: -0.01, 0.41), p=0.056), with an increasing effect of number of pack-years smoked on AF (p=0.024). Conclusions: The results of this study suggest that the existing reference values should be expanded for ethnicity, gender and smoking. These results also indicate that the use of the AGE reader in clinical settings should be used with caution, since the clinical cut-off points are dependent on various factors such as ethnicity that still need to be studied in greater detail.

Autism spectrum disorders (ASD) are a group of neuro-developmental disabilities common in childhood characterized by impairments in social interaction, communication skills, and patterns of activities. Background: The symptoms of ASDs typically are present before age 3 years and often are accompanied by abnormalities in cognitive functioning, learning, attention, and sensory processing. A formal diagnosis is generally not made until the age of five. The vast majority of cases of autism are idiopathic, the best scientific evidence available to us today points toward a potential for various combinations of factors, namely multiple genetic components and certain environmental and socio-economic factors. Timing of exposure during the child's development (before, during or after birth) may also play a role in the development or final presentation of the disorder. Objectives: To estimate the effect of certain socio-economic and other risk factors on the prevalence of autism in Qatar. Methods: The target population for this study are children diagnosed with autism attending the Shafallah Center for children with special needs. Clinical evaluation is conducted by a developmental psychologist, and/or pediatrician, it includes a medical, developmental, and behavioral history; a standard physical and neurological examination, In addition, the Autism Diagnostic Interview (ADI-R), and Autism Diagnostic Observation Schedule-G (ADOS-G) will be administered. Results: Preliminary analysis of 171 subjects showed the highest prevalence among age group 7-14 years (61%). Male/female ratio was 82%/18%, which is around 5/1. Other factors like consanguinity, education, and family income found to have an effect on the prevalence of the disease in Qatar. Conclusions: Obtaining a reliable estimate is important in planning for providing the best health care and educational services needed to improve the overall outcome of autism in Qatar.

Umbilical cord blood (UCB) is an attractive source of hematopoietic stem cells (HSCs). However, the number of HSCs in UBC remains limited and the attempts to amplify them in-vitro remain of poor efficiency. Several publications document amplification of HSC/progenitors with endothelial or mesenchymal cells, but the lack of homogeneity in culture conditions or HSC qualification impairs direct comparison of these results. Therefore, we compared the possible HSCs amplification using placental mesenchymal progenitors and Akt-activated umbilical vein endothelial cells. After HSCs isolation from UCB (defined as Lin-CD45-CD34+CD38-CD90+) on confluent feeder layer, the number of total cells and HSCs were monitored over 21 days. We demonstrate important cellular expansion on both niches. Most of the expanded cells were differentiated when characterized by flow cytometry. Only endothelial cells could trigger HSCs amplification with a pick at 14 days. Those amplified HSCs were able to differentiate in all cell lineages as attested by colony forming assays. Mesenchymal progenitors mainly triggered the amplification of CD38+ cells previously defined as precursors. A competitive assay demonstrated that HSCs had an in-vitro preference to interact with endothelial cells. Cytokines and transcriptomic analysis of our feeders indicate the mechanisms involved in HSC amplification and differentiation in both cases.

Background and Objectives: In the past 40 years Oman has witnessed remarkable social and economic growth, which is best reflected in the well-organized and efficient healthcare system. It has been anticipated that a change in disease pattern would be brought about by improvement in the quality of life, comprehensive healthcare facilities and successful control of communicable diseases. Although genetic and congenital disorders are increasingly observed in medical practice, there were no convincing available data to support these observations. The study was performed over the years 2010-2012 in order to assess the population needs for genetic services. Methods: A population-based study was conducted with detailed assessment and interviews of more than 3000 Omani mothers. Randomization was performed by counting the number of nationals in each geographical area and places of residence, and interviewing one mother per 600 Omani nationals in all geographic locations throughout the country. The collected data were verified with available hospital records and reports and analyzed. Results and Conclusions The population-based study revealed that 10% of Omani mothers reported congenital and genetic disorders causing morbidity in their offspring. Congenital disorders with mental disability accounted for over half of the reported morbidity, and a third of these reports had familiar recurrence. Morbidity and mortality from congenital and genetic causes that have been derived from this study reflect the situation in traditional Muslim community with high rate of inbreeding where communicable diseases were successfully controlled, and prevention measures are still in a preparatory phase. The current study confirmed that congenital and genetic conditions are the major contributors to childhood morbidity and mortality, and handicap prevalence in the Sultanate of Oman, indicating the need to prioritize future healthcare and planning, in view of their significant financial, social and research relevance.

Background: Prostate cancer is the most common malignancy in many industrialized countries and the second cause of cancer-related death in Europe and the United States. The incidence of the disease has been increasing in Arab populations. Large databases focused on genetic susceptibility to prostate cancer have been accumulated from population studies of different ancestries, including Europeans and African-Americans. Arab populations, however, have been only rarely studied. Genetic susceptibility to prostate cancer differs significantly across ethnicities. It would be of interest to identify the common alleles associated with prostate cancer risk in Arab population. Methods: With Affymetrix Genome-Wide Human SNP Array 6.0, we conducted a genome-wide association study (GWAS) in which 534,781 single nucleotide polymorphisms (SNPs) were genotyped in a Tunisian cohort of 92 cases with prostate cancer and 131 age-matched controls. Then we extended the study to evaluate promising associations of 11 SNPs, identified by GWAS, in a cohort of individuals of Arab ancestry living in Qatar and Saudi Arabia (155 cases and 182 controls) using TaqMan® SNP Genotyping Assays. Results: We identified 3 consecutive regions significantly associated with prostate cancer risk on chromosome 9, 17 and 22, including 18 SNPs (P=8.52×10-5 to P=2.18×10-6) in the Tunisian population. Three previous reported common loci associated with prostate cancer at 17q21 containing STAT3 gene in the Caucasian population were confirmed. Two novel chromosomal regions associated with prostate cancer contain candidate susceptibility genes: SMARCA2, LOC646851and SUN2 were revealed. Conclusion: Our findings, identifying novel GWAS prostate cancer susceptibilty loci, indicate that prostate cancer genetic risk factors could be ethnic specific.

Background: Cytogenetics of solid tumors in general is negatively affected by culturing artifacts such as preferential clonal expansion and introduction of chromosomal rearrangements, thus making it difficult to discriminate between primary changes and secondary events. Considering that chromosomal instability is a hallmark of cancer even at the precursor cancer level, it is of a great importance to devise a method of comprehensive chromosome analysis of cancer and cancer precursor cells that can bypass culturing problems as much as possible and that can provide informative karyograms of primary tumors at non-mitosis stages of the cell cycle. Consequently, we have developed a novel approach using chemically induced premature chromosome condensation (PCC) method and combined it with the technique of whole genome chromosome painting assay (M-FISH), in which chromosomal constitution of cancer cells could be detected independent of mitosis and without culture artifact within just three hours after receiving the biopsies. Objectives: 1. To perform karyotype analysis; 2. To define stage specificity; 3. To detect hall-mark(s) of cancer of different origins; 4. To improve therapy regimen by elucidating sensitivity of tumors and patients' lymphocytes to radiation and/or cytostatic drugs. Methods: Chemically-induced PCC method was applied on primary tumors (such as, cervical-, head and neck-, breast- and prostate-cancer) to generate metaphases. Afterwards, these preparations underwent M-FISH. Results: This unique/novel combined assay can give for the first time the possibility to assess genetic instability by detecting spontaneously occurring chromosomal instability (structural and numerical) in different types of primary tumors, as well as adjacent normal cells. It may open up the possibility to detect biomarker(s) for specific type of tumors, that can be used as screening tests. Furthermore, the sensitivity of specific types of tumors as well as patient's normal lymphocytes to cytostatic drugs and/or radiation can be determined. Conclusion: The unique combined state of the art assay we developed can lead to detect biomarker(s) for specific type of cancer. Moreover, for the first time therapy regimen can be individually designed based upon detecting tumors as well as patient's lymphocytes sensitivity.

Objective: To evaluate semen parameters and measure serum follicle-stimulating hormone (FSH), luteinizing hormone (LH), and testosterone (T) concentrations before and 7 days after packed red cell transfusion (PCTx) in adults with sickle cell disease (SCD). Methods: This prospective study investigated 18 young adults with transfusion-dependent SCD, aged 20.7 +/- 2.88 years, with full pubertal development (Tanner's stage 5) (euogonadal), and capacity to ejaculate. Their serum ferritin levels were 1488 +/- 557ng/ml. Basal serum concentrations of FSH, LH, T and IGF-I were evaluated before and 7 days after packed red cell transfusion (PCTx). We studied the effect of PCTx on semen parameters and the endocrine functions in these 18 patients with SCD. Results: Following PCTx, a significant increase of Hb from 8.5 +/- 1.17 g/dl to 10.5 +/- 0.4 g/dl was associated with increased testosterone (12.3 +/- 1.24 nmol/L to 14.23 +/- 1.22nmol/L and gonadotropin concentrations. Total sperm count increased significantly from 87.4 +/- 24.6 million/ml to 146.2 +/- 51.25 million/ml total progressive sperm motility (TPM) from 40.8 +/- 11.1 million/ml to 93.4 +/- 38.3 million/ml, and rapid progressive sperm motility progressive motility (RPM) increased from 29.26 +/- 8.75 million/ml to 67.4 +/- 29 million/ml. After PCTx the total sperm count, TPM and RPM were significantly higher in the ETx group versus the TTx group. Before and after PCTx, testosterone concentrations were correlated significantly with sperm total count, volume, TPM and RPM (r= 0.53, 0.55, 0.42 and 0.38 respectively, p= 0.01). Before and after PCTx, hemoglobin concentrations were correlated significantly with sperm count, TPM, RPM and percentage of sperms with normal morphology (r= 0.60, 0.69, 0.66 and 0.86 respectively, p <0.001) Conclusion: Our study suggests that in males with SCD, blood transfusion associated with significant acute enhancement of sperm parameters and with an increased concentration of testosterone, LH and FSH. Improvements of sperm parameters were significantly higher in the exchange transfusion (ETx) group versus the top-up (TTx) group. These 'acute' effects on spermatogenesis are reached with an unknown mechanism(s) and suggest a number of pathways that need further human and/or experimental studies.

Obesity has increased at an alarming rate over the past three decades. It is likely to be caused by increased caloric intake combined with genetic predisposing factors and is a major risk factor for type 2 diabetes (T2D) and cardiovascular disorders. Obesity is linked to chronic inflammation, which was proposed as a cause for insulin resistance and T2D. However, the molecular mechanism of obesity driven T2D still lacks knowledge and thus is inaccessible to therapeutic intervention. In our research, we are exploring the molecular mechanism underpinning this association and aims to identify novel therapeutic targets for the rapidly growing population of obese individuals living with diabetes. The obese state creates a microenvironment within the adipose tissue that is susceptible to the accumulation of reactive oxygen species (ROS). Accumulated ROS causes oxidation of macromolecules such as DNA. Oxidative DNA damage encompasses a variety of DNA lesions such as DNA single-strand breaks (SSBs) and DNA double-strand breaks (DSBs) including non-telomeric and telomeric DSBs. Broken or dysfunctional telomeres have been hypothesized as a cause of cellular senescence, a state of irreversible cell cycle arrest observed in aging illnesses and even in obesity with accumulation of senescent adipocytes. A major player in detecting and defending against accumulation of oxidative DNA damage is PARP-1. PARP-1 activation has been shown to be associated to diabetes and obesity. We are exploring the role of PARP-1 in ROS-induced senescent and inflamed adipocytes by using a combination of biochemical, molecular and cell biological assays in primary human and mice adipocytes treated with hydrogen peroxide (H₂O₂) as a source of ROS.