Genome-wide association study of migraine implicates a common susceptibility variant on 8q22.1

Migraine is a common episodic neurological disorder, typically presenting with recurrent attacks of severe headache and autonomic dysfunction. Apart from rare monogenic subtypes, no genetic or molecular markers for migraine have been convincingly established. We identified the minor allele of rs1835740 on chromosome 8q22.1 to be associated with migraine (P = 5.38 x 10(-)(9), odds ratio = 1.23, 95% CI 1.150-1.324) in a genome-wide association study of 2,731 migraine cases ascertained from three European headache clinics and 10,747 population-matched controls. The association was replicated in 3,202 cases and 40,062 controls for an overall meta-analysis P value of 1.69 x 10(-)(1)(1) (odds ratio = 1.18, 95% CI 1.127-1.244). rs1835740 is located between MTDH (astrocyte elevated gene 1, also known as AEG-1) and PGCP (encoding plasma glutamate carboxypeptidase). In an expression quantitative trait study in lymphoblastoid cell lines, transcript levels of the MTDH were found to have a significant correlation to rs1835740 (P = 3.96 x 10(-)(5), permuted threshold for genome-wide significance 7.7 x 10(-)(5). To our knowledge, our data establish rs1835740 as the first genetic risk factor for migraine.

Sequence variants in three loci influence monocyte counts and erythrocyte volume

Blood cells participate in vital physiological processes, and their numbers are tightly regulated so that homeostasis is maintained. Disruption of key regulatory mechanisms underlies many blood-related Mendelian diseases but also contributes to more common disorders, including atherosclerosis. We searched for quantitative trait loci (QTL) for hematology traits through a whole-genome association study, because these could provide new insights into both hemopoeitic and disease mechanisms. We tested 1.8 million variants for association with 13 hematology traits measured in 6015 individuals from the Australian and Dutch populations. These traits included hemoglobin composition, platelet counts, and red blood cell and white blood cell indices. We identified three regions of strong association that, to our knowledge, have not been previously reported in the literature. The first was located in an intergenic region of chromosome 9q31 near LPAR1, explaining 1.5% of the variation in monocyte counts (best SNP rs7023923, p=8.9x10(-14)). The second locus was located on chromosome 6p21 and associated with mean cell erythrocyte volume (rs12661667, p=1.2x10(-9), 0.7% variance explained) in a region that spanned five genes, including CCND3, a member of the D-cyclin gene family that is involved in hematopoietic stem cell expansion. The third region was also associated with erythrocyte volume and was located in an intergenic region on chromosome 6q24 (rs592423, p=5.3x10(-9), 0.6% variance explained). All three loci replicated in an independent panel of 1543 individuals (p values=0.001, 9.9x10(-5), and 7x10(-5), respectively). The identification of these QTL provides new opportunities for furthering our understanding of the mechanisms regulating hemopoietic cell fate.

Impacts of a fuel oil spill on seagrass meadows in a subtropical port, Gladstone, Australia - The value of long-term marine habitat monitoring in high risk areas.

We used an established seagrass monitoring programme to examine the short and longer-term impacts of an oil spill event on intertidal seagrass meadows. Results for potentially impacted seagrass areas were compared with existing monitoring data and with control seagrass meadows located outside of the oil spill area. Seagrass meadows were not significantly affected by the oil spill. Declines in seagrass biomass and area 1 month post-spill were consistent between control and impact meadows. Eight months post-spill, seagrass density and area increased to be within historical ranges. The declines in seagrass meadows were likely attributable to natural seasonal variation and a combination of climatic and anthropogenic impacts. The lack of impact from the oil spill was due to several mitigating factors rather than a lack of toxic effects to seagrasses. The study demonstrates the value of long-term monitoring of critical habitats in high risk areas to effectively assess impacts.

Integrating sorghum whole genome sequence information with a compendium of sorghum QTL studies reveals uneven distribution of QTL and of gene-rich regions with significant implications for crop improvement.

A comprehensive analysis was conducted using 48 sorghum QTL studies published from 1995 to 2010 to make information from historical sorghum QTL experiments available in a form that could be more readily used by sorghum researchers and plant breeders. In total, 771 QTL relating to 161 unique traits from 44 studies were projected onto a sorghum consensus map. Confidence intervals (CI) of QTL were estimated so that valid comparisons could be made between studies. The method accounted for the number of lines used and the phenotypic variation explained by individual QTL from each study. In addition, estimated centimorgan (cM) locations were calculated for the predicted sorghum gene models identified in Phytozome (JGI GeneModels SBI v1.4) and compared with QTL distribution genome-wide, both on genetic linkage (cM) and physical (base-pair/bp) map scales. QTL and genes were distributed unevenly across the genome. Heterochromatic enrichment for QTL was observed, with approximately 22% of QTL either entirely or partially located in the heterochromatic regions. Heterochromatic gene enrichment was also observed based on their predicted cM locations on the sorghum consensus map, due to suppressed recombination in heterochromatic regions, in contrast to the euchromatic gene enrichment observed on the physical, sequence-based map. The finding of high gene density in recombination-poor regions, coupled with the association with increased QTL density, has implications for the development of more efficient breeding systems in sorghum to better exploit heterosis. The projected QTL information described, combined with the physical locations of sorghum sequence-based markers and predicted gene models, provides sorghum researchers with a useful resource for more detailed analysis of traits and development of efficient marker-assisted breeding strategies.

Sensory Systems in Sawfishes. 2. The Lateral Line.

The lateral line system allows elasmobranchs to detect hydrodynamic movements in their close surroundings. We examined the distribution of pit organs and lateral line canals in 4 species of sawfish (Anoxypristis cuspidata, Pristis microdon, P. clavata and P. zijsron). Pit organs could only be located in A. cuspidata, which possesses elongated pits that are lined by dermal denticles. In all 4 pristid species, the lateral line canals are well developed and were separated into regions of pored and non-pored canals. In all species the tubules that extend from pored canals form extensive networks. In A. cuspidata, P. microdon and P. clavata, the lateral line canals on both the dorsal and ventral surfaces of the rostrum possess extensively branched and pored tubules. Based on this morphological observation, we hypothesized that these 3 species do not use their rostrum to search in the substrate for prey as previously assumed. Other batoids that possess lateral line canals adapted to perceive stimuli produced by infaunal prey possess non-pored lateral line canals, which also prevent the intrusion of substrate particles. However, this hypothesis remains to be tested behaviourally in pristids. Lateral line canals located between the mouth and the nostrils are non-pored in all 4 species of sawfish. Thus this region is hypothesized to perceive stimuli caused by direct contact with prey before ingestion. Lateral line canals that contain neuromasts are longest in P. microdon, but canals containing neuromasts along the rostrum are longest in A. cuspidata.

Sensory Systems in Sawfishes. 1. The Ampullae of Lorenzini

The distribution and density of the ampullary electroreceptors in the skin of elasmobranchs are influenced by the phylogeny and ecology of a species. Sensory maps were created for 4 species of pristid sawfish. Their ampullary pores were separated into pore fields based on their innervation and cluster formation. Ventrally, ampullary pores are located in 6 areas (5 in Pristis microdon), covering the rostrum and head to the gills. Dorsally, pores are located in 4 areas (3 in P. microdon), which cover the rostrum, head and may extend slightly onto the pectoral fins. In all species, the highest number of pores is found on the dorsal and ventral sides of the rostrum. The high densities of pores along the rostrum combined with the low densities around the mouth could indicate that sawfish use their rostrum to stun their prey before ingesting it, but this hypothesis remains to be tested. The directions of ampullary canals on the ventral side of the rostrum are species specific. P. microdon possesses the highest number of ampullary pores, which indicates that amongst the study species this species is an electroreception specialist. As such, juvenile P. microdon inhabit low-visibility freshwater habitats.

Wet season resting - economic insights from scenario modelling

Pasture degradation, particularly that attributable to overgrazing, is a significant problem across the northern Australian rangelands. Although grazing studies have identified the scope for wet season resting strategies to be used to rehabilitate degraded pastures, the economic outcome of these strategies has not been extensively demonstrated. An exploratory study of the prospective economic value of wet season resting is presented using an economic simulation model of a 28000 ha beef enterprise located in the Charters Towers region of north-eastern Australia to explore seven hypothetical scenarios centred on the projected performance of a wet season resting strategy. A series of 20-year simulations for a range of pasture recovery profiles, stocking capacity, animal productivity responses, beef prices and agistment options are compared with a baseline scenario of taking no action. Estimates of the net present value of the 20-year difference in total enterprise gross margins between the various resting options and the 'do nothing' option identify that wet season resting can offer a positive economic return for the range of scenarios examined, although this is contingent on the assumptions that are made concerning the trajectories of change in carrying capacity and animal productivity. Some implications for management and policy making to support the practical implementation of wet season resting strategies are discussed.

The Role of Genes on Chromosome Locus 4q12 in Human Central Nervous System Tumors

Human central nervous system (CNS) tumors are a heterogeneous group of tumors occurring in brain, brainstem and spinal cord. Malignant gliomas (astrocytic and oligodendroglial tumors), which arise from the neuroepithelial cells are the most common CNS neoplasms in human. Malignant gliomas are highly aggressive and invasive tumors, and have a very poor prognosis. The development and progression of gliomas involve a stepwise accumulation of genetic alterations that generally affect either signal transduction pathways activated by receptor tyrosine kinases (RTKs), or cell cycle arrest pathways. Constitutive activation or deregulated signaling by RTKs is caused by gene amplification, overexpression or mutations. The aberrant RTK signaling results in turn in the activation of several downstream pathways, which ultimately lead to malignant transformation and tumor proliferation. Many genetic abnormalities implicated in nervous system tumors involve the genes located at the chromosomal region 4q12. This locus harbors the receptor tyrosine kinases KIT, PDGFRA and VEGFR2, and other genes (REST, LNX1) with neural function. Gene amplification and protein expression of KIT, PDGFRA, and VEGFR2 was studied using clinical tumor material. REST and LNX1, as well as NUMBL, the interaction partner of LNX1, were studied for their gene mutations and amplifications. In our studies, amplification of LNX1 was associated with KIT and PDGFRA amplification in glioblastomas, and coamplification of KIT, PDGFRA and VEGFR2 was detected in medulloblastomas and CNS primitive neuroectodermal tumors. PDGFRA amplification was also correlated with poor overall survival. Coamplification of KIT, PDGFRA and VEGFR2 was observed in a subset of human astrocytic and oligodendroglial tumors. We suggest that genes at 4q12 could be a part of a larger amplified region, which is deregulated in gliomas, and could be used as a prognostic marker of tumorigenic process. The signaling pathways activated due to gene amplifications, activating gene mutations, and overexpressed proteins may be useful as therapeutic targets for glioma treatment. This study also includes the characterization of KIT overexpressing astrocytes, analyzed by various in vitro functional assays. Our results show that overexpression of KIT in mouse astrocytes promotes cell proliferation and anchorage-independent growth, as well as phenotypic changes in the cells. Furthermore, the increased proliferation is partly inhibited by imatinib, a small molecule inhibitor of KIT. These results suggest that KIT may play a role in astrocyte growth regulation, and might have an oncogenic role in brain tumorigenesis. Elucidation of the altered signaling pathways due to specific gene amplifications, activating gene mutations, and overexpressed proteins may be useful as therapeutic targets for glioma treatment.

Genetic alterations in microsatellite-unstable colorectal cancer

Colorectal cancer (CRC) is the third most common cancer in Finland. Of all CRC tumors, 15% display microsatellite-instability (MSI) caused by defective cellular mismatch repair. Cells displaying MSI accumulate a high number of mutations genome-wide, especially in short repeat areas, microsatellites. When targeting genes essential for cell growth or death, MSI can promote tumorigenesis. In non-coding areas, microsatellite mutations are generally considered as passenger events. Since the discovery of MSI and its linkage to cancer, more that 200 genes have been investigated for a role in MSI tumorigenesis. Although various criteria have been suggested for MSI target gene identification, the challenge has been to distinguish driver mutations from passenger mutations. This study aimed to clarify these key issues in the research field of MSI cancer. Prior to this, background mutation rate in MSI cancer has not been studied in a large-scale. We investigated the background mutation rate in MSI CRC by analyzing the spectrum of microsatellite mutations in non-coding areas. First, semenogelin I was studied for a possible role in MSI carcinogenesis. The intronic T9 repeat of semenogelin I was frequently mutated but no evidence for selection during tumorigenesis was obtained. Second, a sequencing approach was utilized to evaluate the general background mutation rate in MSI CRC. Both intronic and intergenic repeats harbored extremely high mutation rates of ≤ 87% and intergenic repeats were more unstable than the intronic repeats. As mutation rates of presumably neutral microsatellites can be high in MSI CRC in the absence of apparent selection pressure, high mutation frequency alone is not sufficient evidence for identification of driver MSI target genes. Next, an unbiased approach was designed to identify the mutatome of MSI CRC. By combining expression array data and a database search we identified novel genes possibly related to MSI CRC carcinogenesis. One of the genes was studied further. In the functional analysis this gene was observed to cause an abnormal cancer-prone cellular phenotype, possibly through altered responses to DNA damage. In our recent study, smooth muscle myosin heavy chain 11 (MYH11) was identified as a novel MSI CRC gene. Additionally, MYH11 has a well established role in acute myeloid leukemia (AML) through an oncogenic fusion protein CBFB-MYH11. We investigated further the role of MYH11 in AML by sequencing. Three novel missense variants of MYH11 were identified. None of the variants were present in the population-based control material. One of the identified variants, V71A, lies in the N-terminal SH3-like domain of MYH11 of unknown function. The other two variants, K1059E and R1792Q are located in the coil-coiled myosin rod essential for the regulation and filament formation of MYH11. The variant K1059E lies in the close proximity of the K1044N that has been functionally assessed in our earlier work of CRC and has been reported to cause total loss of MYH11 protein regulation. As the functional significance of the three novel variants examined in this work remains unknown, future studies should clarify the further role of MYH11 in AML leukaemogenesis and in other malignancies.

Genomic and functional profiling of gastric cancer

Helicobacter pylori infection is a risk factor for gastric cancer, which is a major health issue worldwide. Gastric cancer has a poor prognosis due to the unnoticeable progression of the disease and surgery is the only available treatment in gastric cancer. Therefore, gastric cancer patients would greatly benefit from identifying biomarker genes that would improve diagnostic and prognostic prediction and provide targets for molecular therapies. DNA copy number amplifications are the hallmarks of cancers in various anatomical locations. Mechanisms of amplification predict that DNA double-strand breaks occur at the margins of the amplified region. The first objective of this thesis was to identify the genes that were differentially expressed in H. pylori infection as well as the transcription factors and signal transduction pathways that were associated with the gene expression changes. The second objective was to identify putative biomarker genes in gastric cancer with correlated expression and copy number, and the last objective was to characterize cancers based on DNA copy number amplifications. DNA microarrays, an in vitro model and real-time polymerase chain reaction were used to measure gene expression changes in H. pylori infected AGS cells. In order to identify the transcription factors and signal transduction pathways that were activated after H. pylori infection, gene expression profiling data from the H. pylori experiments and a bioinformatics approach accompanied by experimental validation were used. Genome-wide expression and copy number microarray analysis of clinical gastric cancer samples and immunohistochemistry on tissue microarray were used to identify putative gastric cancer genes. Data mining and machine learning techniques were applied to study amplifications in a cross-section of cancers. FOS and various stress response genes were regulated by H. pylori infection. H. pylori regulated genes were enriched in the chromosomal regions that are frequently changed in gastric cancer, suggesting that molecular pathways of gastric cancer and premalignant H. pylori infection that induces gastritis are interconnected. 16 transcription factors were identified as being associated with H. pylori infection induced changes in gene expression. NF-κB transcription factor and p50 and p65 subunits were verified using elecrophoretic mobility shift assays. ERBB2 and other genes located in 17q12- q21 were found to be up-regulated in association with copy number amplification in gastric cancer. Cancers with similar cell type and origin clustered together based on the genomic localization of the amplifications. Cancer genes and large genes were co-localized with amplified regions and fragile sites, telomeres, centromeres and light chromosome bands were enriched at the amplification boundaries. H. pylori activated transcription factors and signal transduction pathways function in cellular mechanisms that might be capable of promoting carcinogenesis of the stomach. Intestinal and diffuse type gastric cancers showed distinct molecular genetic profiles. Integration of gene expression and copy number microarray data allowed the identification of genes that might be involved in gastric carcinogenesis and have clinical relevance. Gene amplifications were demonstrated to be non-random genomic instabilities. Cell lineage, properties of precursor stem cells, tissue microenvironment and genomic map localization of specific oncogenes define the site specificity of DNA amplifications, whereas labile genomic features define the structures of amplicons. These conclusions suggest that the definition of genomic changes in cancer is based on the interplay between the cancer cell and the tumor microenvironment.

Rural energy consumption patterns - A field study

This paper is a condensed version of the final report of a detailed field study of rural energy consumption patterns in six villages located west of Bangalore in the dry belt of Karnataka State in India. The study was carried out in two phases; first, a pilot study of four villages and second, the detailed study of six villages, the populations of which varied from around 350 to about 950. The pilot survey ended in late 1976, and most of the data was collected for the main project in 1977. Processing of the collected data was completed in 1980. The aim was to carry out a census survey, rather than a sample study. Hence, considerable effort was expended in production of both a suitable questionnaire, ensuring that all respondents were contacted, and devising methods which would accurately reflect the actual energy use in various energy-utilising activities. In the end, 560 households out of 578 (97%) were surveyed. The following ranking was found for the various energy sources in order of average percentage contribution to the annual total energy requirement: firewood, 81A·6%; human energy, 7A·7%; animal energy, 2A·7%; kerosene, 2A·1%; electricity, 0A·6% and all other sources (rice husks, agro-wastes, coal and diesel fuel), 5A·3%. In other words commercial fuels made only a small contribution to the overall energy use. It should be noted that dung cakes are not burned in this region. The average energy use pattern, sector by sector, again on a percentage basis, was as follows: domestic, 88A·3%; industry, 4A·7%; agriculture, 4A·3%; lighting, 2A·2% and transport, 0A·5%. The total annual per capita energy consumption was 12A·6 A± 1A·2 GJ, giving an average annual household consumption of around 78A·6 GJ.

Electron- Spin-Resonance studies of phase transitions in double propionates

Detailed ESR investigations of Mn2+ substituting for Ca2+ in Ca2Sr(C2H5COO)6, (DSP) and Ca2Pb(C2H5COO)6, (DLP) and Ca2Ba(C2H5COO)6, (DBP), in single crystals and powders, over the temperature range from 300°C to -180°C have been carried out to study the successive phase transitions in these compounds. Spectra have been analyzed in terms of axial spin Hamiltonians and the temperature dependences of the parameters studied. Across the I-II transition, new physically and chemically inequivalent sites appear indicating the disappearance of the diad axes on which the propionate groups are located, bringing out the connection between the motional states of the propionate groups and the occurrence of ferroelectricity. The II-III transition also causes chemically inequivalent sites to develop, indicating that the transitions may not be isomorphous as believed previously. Similarities and dissimilarities of the ESR spectra of DLP, DSP and DBP are discussed in relation to the phase transitions.

Quantitative Trait Loci for CD4:CD8 Lymphocyte Ratio Are Associated with Risk of Type 1 Diabetes and HIV-1 Immune Control

Abnormal expansion or depletion of particular lymphocyte subsets is associated with clinical manifestations such as HIV progression to AIDS and autoimmune disease. We sought to identify genetic predictors of lymphocyte levels and reasoned that these may play a role in immune-related diseases. We tested 2.3 million variants for association with five lymphocyte subsets, measured in 2538 individuals from the general population, including CD4+ T cells, CD8+ T cells, CD56+ natural killer (NK) cells, and the derived measure CD4:CD8 ratio. We identified two regions of strong association. The first was located in the major histocompatibility complex (MHC), with multiple SNPs strongly associated with CD4:CD8 ratio (rs2524054, p = 2.1 × 10−28). The second region was centered within a cluster of genes from the Schlafen family and was associated with NK cell levels (rs1838149, p = 6.1 × 10−14). The MHC association with CD4:CD8 replicated convincingly (p = 1.4 × 10−9) in an independent panel of 988 individuals. Conditional analyses indicate that there are two major independent quantitative trait loci (QTL) in the MHC region that regulate CD4:CD8 ratio: one is located in the class I cluster and influences CD8 levels, whereas the second is located in the class II cluster and regulates CD4 levels. Jointly, both QTL explained 8% of the variance in CD4:CD8 ratio. The class I variants are also strongly associated with durable host control of HIV, and class II variants are associated with type-1 diabetes, suggesting that genetic variation at the MHC may predispose one to immune-related diseases partly through disregulation of T cell homeostasis.

A case study on the economics of overhead irrigation in the lower Burdekin

Converting from an existing irrigation system is often seen as high risk by the land owner. The significant financial investment and the long period over which the investment runs is also complicated by the uncertainty associated with long term input costs (such as energy), crop production, and the continually evolving natural resource management rules and policy. Irrigation plays a pivotal part in the Burdekin sugarcane farming system. At present the use of furrow irrigation is by far the most common form due to the ease of use, relatively low operating cost and well established infrastructure currently in place. The Mulgrave Area Farmer Integrated Action (MAFIA) grower group, located near Clare in the lower Burdekin region, identified the need to learn about sustainable farming systems with a focus on the environment, social and economic implications. In early 2007, Hesp Faming established a site to investigate the use of overhead irrigation as an alternative to furrow irrigation and its integration with new farming system practices, including Green Cane Trash Blanketing (GCTB). Although significant environmental and social benefits exist, the preliminary investment analysis indicates that the Overhead Low Pressure (OHLP) irrigation system is not adding financial value to the Hesp Farming business. A combination of high capital costs and other offsetting factors resulted in the benefits not being fully realised. A different outcome is achieved if Hesp Farming is able to realise value on the water saved, with both OHLP irrigation systems displaying a positive NPV. This case study provides a framework to further investigate the economics of OHLP irrigation in sugarcane and it is anticipated that with additional data a more definitive outcome will be developed in the future.

The role of tillage, fertiliser and forage species in sustaining dairying based on crops in southern Queensland 2. Double-crop and summer sole-crop systems.

Dairy farms located in the subtropical cereal belt of Australia rely on winter and summer cereal crops, rather than pastures, for their forage base. Crops are mostly established in tilled seedbeds and the system is vulnerable to fertility decline and water erosion, particularly over summer fallows. Field studies were conducted over 5 years on contrasting soil types, a Vertosol and Sodosol, in the 650-mm annual-rainfall zone to evaluate the benefits of a modified cropping program on forage productivity and the soil-resource base. Growing forage sorghum as a double-crop with oats increased total mean annual production over that of winter sole-crop systems by 40% and 100% on the Vertosol and Sodosol sites respectively. However, mean annual winter crop yield was halved and overall forage quality was lower. Ninety per cent of the variation in winter crop yield was attributable to fallow and in-crop rainfall. Replacing forage sorghum with the annual legume lablab reduced fertiliser nitrogen (N) requirements and increased forage N concentration, but reduced overall annual yield. Compared with sole-cropped oats, double-cropping reduced the risk of erosion by extending the duration of soil water deficits and increasing the time ground was under plant cover. When grown as a sole-crop, well fertilised forage sorghum achieved a mean annual cumulative yield of 9.64 and 6.05 t DM/ha on the Vertosol and Sodosol, respectively, being about twice that of sole-cropped oats. Forage sorghum established using zero-tillage practices and fertilised at 175 kg N/ha. crop achieved a significantly higher yield and forage N concentration than did the industry-standard forage sorghum (conventional tillage and 55 kg N/ha. crop) on the Vertosol but not on the Sodosol. On the Vertosol, mean annual yield increased from 5.65 to 9.64 t DM/ha (33 kg DM/kg N fertiliser applied above the base rate); the difference in the response between the two sites was attributed to soil type and fertiliser history. Changing both tillage practices and N-fertiliser rate had no affect on fallow water-storage efficiency but did improve fallow ground cover. When forage sorghum, grown as a sole crop, was replaced with lablab in 3 of the 5 years, overall forage N concentration increased significantly, and on the Vertosol, yield and soil nitrate-N reserves also increased significantly relative to industry-standard sorghum. All forage systems maintained or increased the concentration of soil nitrate-N (0-1.2-m soil layer) over the course of the study. Relative to sole-crop oats, alternative forage systems were generally beneficial to the concentration of surface-soil (0-0.1 m) organic carbon and systems that included sorghum showed most promise for increasing soil organic carbon concentration. We conclude that an emphasis on double-or summer sole-cropping rather than winter sole-cropping will advantage both farm productivity and the soil-resource base.