Water resources in Saudi Arabia with particular reference to Tihama Asir province

Although Saudi Arabia has made great progress in almost all aspects of development, the availability of water has remained problematic. It is, therefore, important to study water resources, and also the increasing water demands in Saudi Arabia in general, and in Tihama Asir in particular. Climate is influenced to a limited extent by the relatively high altitude of the mountains, especially where rainfall is concerned, and an annual average between 30mm and 300mm is recorded. The rate of evaporation, however, is very high, because of the cloudless sky and high temperatures. Runoff occurs only after torrential and monsoon rain, when the wadis collect floodwaters from their many tributaries. Two hundred dams have been built in recent years in an attempt to increase underground water recharge, and to provide potable water in sufficient quantity and quality, and to provide for irrigation. The ground water resources, including the shallow aquifers of wadis and the deep aquifers, could be of greater benefit with the modification of their current use. A large number of desalination plants have been constructed along the Red Sea and the Arabian Gulf to meet the increasing water demands of Saudi Arabia's increasing population. The utilization of treated sewage effluent should be increased to stop the threat of ground water contamination and to reduce the pressure exerted on available resources. Results indicate that modem irrigation methods reach high levels of average field irrigation efficiency, and traditional practices show the highest average levels of energetics performance. Based on water resources and water budget analysis the following recommendations are suggested to the Ministry of Agriculture and Water with regard to water uses in Saudi Arabia and Tihama Asir in particular.- Modernize traditional and intermediate irrigation methods on as many acres as feasible.- Introduce small green-houses on as many private farms as possible.- Develop water resources extension service programmes and training programmes that address fundamental aspects of improving domestic water use, and alternatives to natural and artificial supplies. The developed database can be used as an information source to support future water resource-oriented decision making

Integrated approaches to elucidate the genetic architecture of congenital heart defects

Congenital heart defects (CHD) are structural anomalies affecting the heart, are found in 1% of the population and arise during early stages of embryo development. Without surgical and medical interventions, most of the severe CHD cases would not survive after the first year of life. The improved health care for CHD patients has increased CHD prevalence significantly, and it has been estimated that the population of adults with CHD is growing ~5% per year. Understanding the causes of CHD would greatly help improve our knowledge of the pathophysiology, family counseling and planning and possibly prevention and treatment in the future. Several lines of evidence from humans and animal models have supported a substantial genetic component for CHD. However, gene discovery in CHD has been difficult due to the extreme locus heterogeneity and the lack of a distinct genotype–phenotype correlation. Currently, genetic causes are identified in fewer than 20-­‐30% of the cases, most of which are syndromic while the isolated CHD cases remain largely without explanation. The aim of my thesis was to identify novel or known CHD genes enriched for rare coding genetic variants in isolated CHD cases and learn about the relative performance of different study designs. High-throughput next generation sequencing (NGS) was used to sequence all coding genes (whole exome) coupled with various analytical pipelines and tools to identify candidate genes in different family-based study designs. Since there is no general consensus on the underlying genetic model of isolated CHD, I developed a suite of software tools to enable different family-based exome analyses of de novo and inherited variants (chapter 2) and then piloted these tools in several gene discovery projects where the mode of inheritance was already known to identify previously described and novel pathogenic genes, before applying them to an analysis of families with two or more siblings with CHD. Based on the tools developed in chapter 2, I designed a two-stage study to investigate isolated parent-offspring trios with Tetralogy of Fallot (chapter 3). In the first stage, I used whole exome sequence data from 30 trios to identify genes with de novo coding variants. This analysis identified six de novo loss-of-function and 13 de novo missense variants. Only one gene showed recurrent de novo mutations in NOTCH1, a well known CHD gene that has mostly been associated with left ventricle outflow tract malformations (LVOT). Besides NOTCH1, the de novo analysis identified several possibly pathogenic novel genes such as ZMYM2 and ARHGAP35, that harbor de novo loss-of-function variants (frameshift and stop gain, respectively). In the second stage of the study, I designed custom baits to capture 122 candidate genes for additional sequencing using NGS in a larger sample size of 250 parent-offspring trios with isolated Tetralogy of Fallot and identified six de novo variants in four genes, half of them are loss-of-function variants. Both of NOTCH1 and its ligand JAG1 harbor two additional de novo mutations (two stop gains in NOTCH1 and one missense and a splice donor in JAG1). The analysis showed a strongly significant over-representation of de novo loss-of-function variants in NOTCH1 (P=3.8 ×10-9). Additionally, when compared with 1,080 control trios, NOTCH1 exhibit significant burden of inherited rare missense variant (minor allele frequency < 1% in 1000 genomes) (Fisher exact test, P= 8.8 × 10^­‐05) in about 10% of the isolated Tetralogy of Fallot patients. I also modified the transmission disequilibrium test (TDT) to detect any distortion of rare coding allele transmission from healthy parent to their affected children. This modified TDT test identified ARHGAP35 gene, which exhibits an over-­‐transmission of rare missense variants in children (P=0.025). Although, the p value does not reach a genome-­‐wide significant level after correcting for multiple tests, ARHGAP35 gene has also a de novo stop gain variant in one trio from the primary cohort and recently shown to play a role in cardiomyocyte fate which make it an interesting novel ToF candidate gene for future studies. To assess alternative family-based study design in CHD, I combined the analysis from 13 isolated parent-offspring trios with 112 unrelated index cases of isolated atrioventricular septal defects (AVSD) in chapter 4. Initially, I started with a case/control analysis to test the burden of rare missense variants in cases compared with 5,194 ethnically matching controls and identified the gene NR2F2 (Fisher exact test P=7.7×10-07, odds ratio=54). The de novo analysis in the AVSD trios identified two de novo missense variants in the same gene. NR2F2 encodes a pleiotropic developmental transcription factor, and decreased dosage of NR2F2 in mice has been shown to result in abnormal development of atrioventricular septa. The results from luciferase assays show that all coding sequence variants observed in patients significantly alter the activity of NR2F2 target promoters. My work has identified both known and novel CHD genes enriched for rare coding variants using next-generation sequencing data. I was able to show how using single or combined family-based study designs is an effective approach to study the genetic causes of isolated CHD subtypes. Despite the extreme heterogeneity of CHD, combining NGS data with the proper study design has proved to be an effective approach to identify novel and known CHD genes. Future studies with considerably larger sample sizes are required to yield deeper insights into the genetic causes of isolated CHD.National Guard Health Affairs, Riyadh, Saudi Arabi