Molecular characterisation of environmental enterococci derived from water samples and assessment of associated public health hazards

Enterococci are versatile Gram-positive bacteria that can survive under extreme conditions. Most enterococci are non-virulent and found in the gastrointestinal tract of humans and animals. Other strains are opportunistic pathogens that contribute to a large number of nosocomial infections globally. Epidemiological studies demonstrated a direct relationship between the density of enterococci in surface waters and the risk of swimmer-associated gastroenteritis. The distribution of infectious enterococcal strains from the hospital environment or other sources to environmental water bodies through sewage discharge or other means, could increase the prevalence of these strains in the human population. Environmental water quality studies may benefit from focusing on a subset of Enterococcus spp. that are consistently associated with sources of faecal pollution such as domestic sewage, rather than testing for the entire genus. E. faecalis and E. faecium are potentially good focal species for such studies, as they have been consistently identified as the dominant Enterococcus spp. in human faeces and sewage. On the other hand enterococcal infections are predominantly caused by E. faecalis and E. faecium. The characterisation of E. faecalis and E. faecium is important in studying their population structures, particularly in environmental samples. In developing and implementing rapid, robust molecular genotyping techniques, it is possible to more accurately establish the relationship between human and environmental enterococci. Of particular importance, is to determine the distribution of high risk enterococcal clonal complexes, such as E. faecium clonal complex 17 and E. faecalis clonal complexes 2 and 9 in recreational waters. These clonal complexes are recognized as particularly pathogenic enterococcal genotypes that cause severe disease in humans globally. The Pimpama-Coomera watershed is located in South East Queensland, Australia and was investigated in this study mainly because it is used intensively for agriculture and recreational purposes and has a strong anthropogenic impact. The primary aim of this study was to develop novel, universally applicable, robust, rapid and cost effective genotyping methods which are likely to yield more definitive results for the routine monitoring of E. faecalis and E. faecium, particularly in environmental water sources. To fullfill this aim, new genotyping methods were developed based on the interrogation of highly informative single nucleotide polymorphisms (SNPs) located in housekeeping genes of both E. faecalis and E. faecium. SNP genotyping was successfully applied in field investigations of the Coomera watershed, South-East Queensland, Australia. E. faecalis and E. faecium isolates were grouped into 29 and 23 SNP profiles respectively. This study showed the high longitudinal diversity of E. faecalis and E. faecium over a period of two years, and both human-related and human-specific SNP profiles were identified. Furthermore, 4.25% of E. faecium strains isolated from water was found to correspond to the important clonal complex-17 (CC17). Strains that belong to CC17 cause the majority of hospital outbreaks and clinical infections globally. Of the six sampling sites of the Coomera River, Paradise Point had the highest number of human-related and human-specific E. faecalis and E. faecium SNP profiles. The secondary aim of this study was to determine the antibiotic-resistance profiles and virulence traits associated with environmental E. faecalis and E. faecium isolates compared to human pathogenic E. faecalis and E. faecium isolates. This was performed to predict the potential health risks associated with coming into contact with these strains in the Coomera watershed. In general, clinical isolates were found to be more resistant to all the antibiotics tested compared to water isolates and they harbored more virulence traits. Multi-drug resistance was more prevalent in clinical isolates (71.18% of E. faecalis and 70.3 % of E. faecium) compared to water isolates (only 5.66 % E. faecium). However, tetracycline, gentamicin, ciprofloxacin and ampicillin resistance was observed in water isolates. The virulence gene esp was the most prevalent virulence determinant observed in clinical isolates (67.79% of E. faecalis and 70.37 % of E. faecium), and this gene has been described as a human-specific marker used for microbial source tracking (MST). The presence of esp in water isolates (16.36% of E. faecalis and 19.14% of E. faecium) could be indicative of human faecal contamination in these waterways. Finally, in order to compare overall gene expression between environmental and clinical strains of E. faecalis, a comparative gene hybridization study was performed. The results of this investigation clearly demonstrated the up-regulation of genes associated with pathogenicity in E. faecalis isolated from water. The expression study was performed at physiological temperatures relative to ambient temperatures. The up-regulation of virulence genes demonstrates that environmental strains of E. faecalis can pose an increased health risk which can lead to serious disease, particularly if these strains belong to the virulent CC17 group. The genotyping techniques developed in this study not only provide a rapid, robust and highly discriminatory tool to characterize E. faecalis and E. faecium, but also enables the efficient identification of virulent enterococci that are distributed in environmental water sources.

Measuring reef complexity and rugosity from monocular video bathymetric reconstruction

Topographic structural complexity of a reef is highly correlated to coral growth rates, coral cover and overall levels of biodiversity, and is therefore integral in determining ecological processes. Modeling these processes commonly includes measures of rugosity obtained from a wide range of different survey techniques that often fail to capture rugosity at different spatial scales. Here we show that accurate estimates of rugosity can be obtained from video footage captured using underwater video cameras (i.e., monocular video). To demonstrate the accuracy of our method, we compared the results to in situ measurements of a 2m x 20m area of forereef from Glovers Reef atoll in Belize. Sequential pairs of images were used to compute fine scale bathymetric reconstructions of the reef substrate from which precise measurements of rugosity and reef topographic structural complexity can be derived across multiple spatial scales. To achieve accurate bathymetric reconstructions from uncalibrated monocular video, the position of the camera for each image in the video sequence and the intrinsic parameters (e.g., focal length) must be computed simultaneously. We show that these parameters can be often determined when the data exhibits parallax-type motion, and that rugosity and reef complexity can be accurately computed from existing video sequences taken from any type of underwater camera from any reef habitat or location. This technique provides an infinite array of possibilities for future coral reef research by providing a cost-effective and automated method of determining structural complexity and rugosity in both new and historical video surveys of coral reefs.

Self-calibration of wireless cameras with restricted degrees of freedom

This paper presents an approach for the automatic calibration of low-cost cameras which are assumed to be restricted in their freedom of movement to either pan or tilt movements. Camera parameters, including focal length, principal point, lens distortion parameter and the angle and axis of rotation, can be recovered from a minimum set of two images of the camera, provided that the axis of rotation between the two images goes through the camera’s optical center and is parallel to either the vertical (panning) or horizontal (tilting) axis of the image. Previous methods for auto-calibration of cameras based on pure rotations fail to work in these two degenerate cases. In addition, our approach includes a modified RANdom SAmple Consensus (RANSAC) algorithm, as well as improved integration of the radial distortion coefficient in the computation of inter-image homographies. We show that these modifications are able to increase the overall efficiency, reliability and accuracy of the homography computation and calibration procedure using both synthetic and real image sequences

The 3D kinematics of the single leg flat and decline squat

Background: Pre-participation screening is commonly used to measure and assess potential intrinsic injury risk. The single leg squat is one such clinical screening measure used to assess lumbopelvic stability and associated intrinsic injury risk. With the addition of a decline board, the single leg decline squat (SLDS) has been shown to reduce ankle dorsiflexion restrictions and allowed greater sagittal plane movement of the hip and knee. On this basis, the SLDS has been employed in the Cricket Australia physiotherapy screening protocols as a measure of lumbopelvic control in the place of the more traditional single leg flat squat (SLFS). Previous research has failed to demonstrate which squatting technique allows for a more comprehensive assessment of lumbopelvic stability. Tenuous links are drawn between kinematics and hip strength measures within the literature for the SLS. Formal evaluation of subjective screening methods has also been suggested within the literature. Purpose: This study had several focal points namely 1) to compare the kinematic differences between the two single leg squatting conditions, primarily the five key kinematic variables fundamental to subjectively assess lumbopelvic stability; 2) determine the effect of ankle dorsiflexion range of motion has on squat kinematics in the two squat techniques; 3) examine the association between key kinematics and subjective physiotherapists’ assessment; and finally 4) explore the association between key kinematics and hip strength. Methods: Nineteen (n=19) subjects performed five SLDS and five SLFS on each leg while being filmed by an 8 camera motion analysis system. Four hip strength measures (internal/external rotation and abd/adduction) and ankle dorsiflexion range of motion were measured using a hand held dynamometer and a goniometer respectively on 16 of these subjects. The same 16 participants were subjectively assessed by an experienced physiotherapist for lumbopelvic stability. Paired samples t-tests were performed on the five predetermined kinematic variables to assess the differences between squat conditions. A Bonferroni correction for multiple comparisons was used which adjusted the significance value to p = 0.005 for the paired t-tests. Linear regressions were used to assess the relationship between kinematics, ankle range of motion and hip strength measures. Bivariate correlations between hip strength measures and kinematics and pelvic obliquity were employed to investigate any possible relationships. Results: 1) Significant kinematic differences between squats were observed in dominant (D) and non-dominant (ND) end of range hip external rotation (ND p = <0.001; D p = 0.004) and hip adduction kinematics (ND p = <0.001; D p = <0.001). With the mean angle, only the non-dominant leg observed significant differences in hip adduction (p = 0.001) and hip external rotation (p = <0.001); 2) Significant linear relationships were observed between clinical measures of ankle dorsiflexion and sagittal plane kinematic namely SLFS dominant ankle (p = 0.006; R2 = .429), SLFS non-dominant knee (p = 0.015; R2 = .352) and SLFS non-dominant ankle (p = 0.027; R2 = .305) kinematics. Only the dominant ankle (p = 0.020; R2 = .331) was found to have a relationship with the decline squat. 3) Strength measures had tenuous associations with the subjective assessments of lumbopelvic stability with no significant relationships being observed. 4) For the non-dominant leg, external rotation strength and abduction strength were found to be significantly correlated with hip rotation kinematics (Newtons r = 0.458 p = 0.049; Normalised for bodyweight: r = 0.469; p = 0.043) and pelvic obliquity (normalised for bodyweight: r = 0.498 p = 0.030) respectively for the SLFS only. No significant relationships were observed in the dominant leg for either squat condition. Some elements of the hip strength screening protocols had linear relationships with kinematics of the lower limb, particularly the sagittal plane movements of the knee and ankle. Strength measures had tenuous associations with the subjective assessments of lumbopelvic stability with no significant relationships being observed; Discussion: The key finding of this study illustrated that kinematic differences can occur at the hip without significant kinematic differences at the knee as a result of the introduction of a decline board. Further observations reinforce the role of limited ankle dorsiflexion range of motion on sagittal plane movement of the hip and knee and in turn multiplanar kinematics of the lower limb. The kinematic differences between conditions have clinical implications for screening protocols that employ frontal plane movement of the knee as a guide for femoral adduction and rotation. Subjects who returned stronger hip strength measurements also appeared to squat deeper as characterised by differences in sagittal plane kinematics of the knee and ankle. Despite the aforementioned findings, the relationship between hip strength and lower limb kinematics remains largely tenuous in the assessment of the lumbopelvic stability using the SLS. The association between kinematics and the subjective measures of lumbopelvic stability also remain tenuous between and within SLS screening protocols. More functional measures of hip strength are needed to further investigate these relationships. Conclusion: The type of SLS (flat or decline) should be taken into account when screening for lumbopelvic stability. Changes to lower limb kinematics, especially around the hip and pelvis, were observed with the introduction of a decline board despite no difference in frontal plane knee movements. Differences in passive ankle dorsiflexion range of motion yielded variations in knee and ankle kinematics during a self-selected single leg squatting task. Clinical implications of removing posterior ankle restraints and using the knee as a guide to illustrate changes at the hip may result in inaccurate screening of lumbopelvic stability. The relationship between sagittal plane lower limb kinematics and hip strength may illustrate that self-selected squat depth may presumably be a useful predictor of the lumbopelvic stability. Further research in this area is required.

Larger or broader : performance implications of size and diversity of the knowledge worker's egocentric network. [更多或更广：知识员工个人中心网络大小和 多样性对绩效的影响]

Management scholars and practitioners emphasize the importance of the size and diversity of a knowledge worker's social network. Constraints on knowledge workers’ time and energy suggest that more is not always better. Further, why and how larger networks contribute to valuable outcomes deserves further understanding. In this study, we offer hypotheses to shed insight on the question of the diminishing returns of large networks and the specific form of network diversity that may contribute to innovative performance among knowledge workers. We tested our hypotheses using data collected from 93 R&D engineers in a Sino-German automobile electronics company located in China. Study findings identified an inflection point, confirming our hypothesis that the size of the knowledge worker's egocentric network has an inverted U-shaped effect on job performance. We further demonstrate that network dispersion richness (the number of cohorts that the focal employee has connections to) rather than network dispersion evenness (equal distribution of ties across the cohorts) has more influence on the knowledge worker's job performance. Additionally, we found that the curvilinear effect of network size is fully mediated by network dispersion richness. Implications for future research on social networks in China and Western contexts are discussed.

Sentencing indigenous and non-indigenous women in Western Australia's higher courts : A summary of the main findings of a narrative study

This paper presents the main findings of a narrative examination of higher court sentencing remarks to explore the relationship between Indigeneity and sentencing for female defendants in Western Australia. Using the theoretical framework of focal concerns, we found that key differences in the construction of blameworthiness and risk between the sentencing stories of Indigenous and non-Indigenous female offenders, through the identification of issues such as mental health, substance abuse, familial trauma and community ties. Further, in the sentencing narratives, Indigenous women were viewed differently in terms of social costs of imprisonment.

Social cars : sensing, gathering, sharing and conveying social cues to road users

Intelligent Transport Systems (ITS) resembles the infrastructure for ubiquitous computing in the car. It encompasses a) all kinds of sensing technologies within vehicles as well as road infrastructure, b) wireless communication protocols for the sensed information to be exchanged between vehicles (V2V) and between vehicles and infrastructure (V2I), and c) appropriate intelligent algorithms and computational technologies that process these real-time streams of information. As such, ITS can be considered a game changer. It provides the fundamental basis of new, innovative concepts and applications, similar to the Internet itself. The information sensed or gathered within or around the vehicle has led to a variety of context-aware in-vehicular technologies within the car. A simple example is the Anti-lock Breaking System (ABS), which releases the breaks when sensors detect that the wheels are locked. We refer to this type of context awareness as vehicle/technology awareness. V2V and V2I communication, often summarized as V2X, enables the exchange and sharing of sensed information amongst cars. As a result, the vehicle/technology awareness horizon of each individual car is expanded beyond its observable surrounding, paving the way to technologically enhance such already advanced systems. In this chapter, we draw attention to those application areas of sensing and V2X technologies, where the human (driver), the human’s behavior and hence the psychological perspective plays a more pivotal role. The focal points of our project are illustrated in Figure 1: In all areas, the vehicle first (1) gathers or senses information about the driver. Rather than to limit the use of such information towards vehicle/technology awareness, we see great potential for applications in which this sensed information is then (2) fed back to the driver for an increased self-awareness. In addition, by using V2V technologies, it can also be (3) passed to surrounding drivers for an increased social awareness, or (4), pushed even further, into the cloud, where it is collected and visualized for an increased, collective urban awareness within the urban community at large, which includes all city dwellers.

Load-unloading response of intact and artificially degraded articular cartilage correlated with near infrared (NIR) absorption spectra

The conventional mechanical properties of articular cartilage, such as compressive stiffness, have been demonstrated to be limited in their capacity to distinguish intact (visually normal) from degraded cartilage samples. In this paper, we explore the correlation between a new mechanical parameter, namely the reswelling of articular cartilage following unloading from a given compressive load, and the near infrared (NIR) spectrum. The capacity to distinguish mechanically intact from proteoglycan-depleted tissue relative to the "reswelling" characteristic was first established, and the result was subsequently correlated with the NIR spectral data of the respective tissue samples. To achieve this, normal intact and enzymatically degraded samples were subjected to both NIR probing and mechanical compression based on a load-unload-reswelling protocol. The parameter δ(r), characteristic of the osmotic "reswelling" of the matrix after unloading to a constant small load in the order of the osmotic pressure of cartilage, was obtained for the different sample types. Multivariate statistics was employed to determine the degree of correlation between δ(r) and the NIR absorption spectrum of relevant specimens using Partial Least Squared (PLS) regression. The results show a strong relationship (R(2)=95.89%, p<0.0001) between the spectral data and δ(r). This correlation of δ(r) with NIR spectral data suggests the potential for determining the reswelling characteristics non-destructively. It was also observed that δ(r) values bear a significant relationship with the cartilage matrix integrity, indicated by its proteoglycan content, and can therefore differentiate between normal and artificially degraded proteoglycan-depleted cartilage samples. It is therefore argued that the reswelling of cartilage, which is both biochemical (osmotic) and mechanical (hydrostatic pressure) in origin, could be a strong candidate for characterizing the tissue, especially in regions surrounding focal cartilage defects in joints.

Bargara Pasturage Reserve : Future Visions

Bargara Pasturage Reserve: Future Visions This exhibition showcases the work of Postgraduate Landscape Architecture and final year Undergraduate Civil and Environmental Engineering students in response to issues of sustainability in a coastal wetland known as the Bargara Pasturage Reserve; an exemplar of the many issues facing sensitive coastal places in Queensland today. The 312ha Pasturage Reserve at Bargara is the only biofilter between the pressures of Bargara’s urban and tourism expansion, surrounding sugarcane farming, and the Great Sandy Marine Park, including the largest concentration of nesting marine turtles on the eastern Australian mainland. This ephemeral wetland, while struggling to fulfil its coastal biofiltration function, is also in high demand for passive recreation, and the project partners’ priorities were to meet both of these challenges. The students were required to plan and design for the best balance possible amongst, but not limited to: wetland and coastal ecological health, enhancement of cultural heritage and values, sustainable urban development, and local economic health. To understand these challenges, QUT staff and students met with partners, visited and analysed the Pasturage Reserve, spent time in and around Bargara talking to locals and inviting dialogue with Indigenous representatives and the South Sea Islander community. We then went home to Brisbane to undertake theoretical and technical research, and then worked to produce 11 Strategic Plans, 2 Environmental Management Plans and 33 Detailed Designs. One group of students analysed the Bargara coastal landscape as an historical and ongoing series of conversations between ecological systems, cultural heritage, community and stakeholders. Another group identified the landscape as neither ‘urban,’ ‘rural,’ nor ‘natural,’ instead identifying it metaphorically as a series of layered thematic ‘fields’ such as water, conservation, reconciliation, and educational fields. These landscape analyses became the organising mechanisms for strategic planning. An outstanding Strategic Plan was produced by Zhang, Lemberg and Jensen, entitled Metanoia, which means to ‘make a change as the result of reflection on values’. Three implementation phases of “flow”, “flux”, and “flex” span twenty-five years, and present a vision a coastal and marine research and conservation hub, with a focus on coastal wetland function, turtle habitat and coral reef conservation. An Environmental Management Plan by Brand and Stickland focuses on protecting and improving wetland biodiversity and habitat quality, and increasing hydrological and water quality function; vital in a coastal area of such high conservation value. After the planning phase, students individually developed detailed design proposals responsive to their plans. From Metanoia, Zhang concentrated on wetland access and interpretation, proposing four focal places to form the nucleus of a wider pattern of connectivity, and to encourage community engagement with coastal environmental management and education. Jensen tackled the thorny issue of coastal urban development, proposing a sensitive staged eco-village model which maintains both ecological and recreational connectivity between the wetland and the marine environment. This project offered QUT’s partners many innovative options to inform their future planning. BSC, BMRG and Oceanwatch Australia are currently engaged in the investigation of on-ground opportunities drawing on these options.

The results of acetabular impaction grafting in 129 primary cemented total hip arthroplasties

Between 1995 and 2003, 129 cemented primary THRs were performed using full acetabular impaction grafting to reconstruct acetabular deficiencies. These were classified as cavitary in 74 and segmental in 55 hips. Eighty-one patients were reviewed at mean 9.1 (6.2-14.3) years post-operatively. There were seven acetabular component revisions due to aseptic loosening, and a further 11 cases that had migrated >5mm or tilted >5° on radiological review - ten of which reported no symptoms. Kaplan-Meier analysis of revisions for aseptic loosening demonstrates 100% survival at nine years for cavitary defects compared to 82.6% for segmental defects. Our results suggest that the medium-term survival of this technique is excellent when used for purely cavitary defects but less predictable when used with large rim meshes in segmental defects.

Early years research

Early years research is increasingly concerned with the everyday lives of young children and adults in the cut-and-thrust of early years contexts. It is concerned with what happens in situ, that is, in the everyday lives of those within the context. It is concerned with understanding young children and adults in the contexts of their lives; but it goes beyond understanding to transforming their contexts such that children and adults have the best possible chances, now and in the future. The dual focus of understanding and transforming makes early years research a powerful force for change. This chapter explores key theoretical underpinnings of early years research and presents key aspects of conducting research in ethical and sustainable ways. Early years research, here, refers to research conducted by early years practitioner researchers in the context of their own setting. It may involve research around their own practice and/or research around a particular issue or phenomenon of importance in their setting – the focal point may be children, families or practitioners or combinations thereof. The research may be a seamless part of the daily routine of the setting or may be a discreet project, clearly delineated with a timeframe for commencement and conclusion. The research may be used for ongoing reflection and planning with the setting and/or for dissemination in research reports or scholarly publications.

Repeated intrauterine exposures to inflammatory stimuli attenuated transforming growth factor-β signaling in the ovine fetal lung

Background: Bronchopulmonary dysplasia (BPD) is one of the most common complications after preterm birth and is associated with intrauterine exposure to bacteria. Transforming growth factor-β (TGFβ) is implicated in the development of BPD. Objectives: We hypothesized that different and/or multiple bacterial signals could elicit divergent TGFβ signaling responses in the developing lung. Methods: Time-mated pregnant Merino ewes received an intra-amniotic injection of lipopolysaccharide (LPS) and/or Ureaplasma parvum serovar 3 (UP) at 117 days' and/or 121/122 days' gestational age (GA). Controls received an equivalent injection of saline and or media. Lambs were euthanized at 124 days' GA (term = 150 days' GA). TGFβ1, TGFβ2, TGFβ3, TGFβ receptor (R)1 and TGFβR2 protein levels, Smad2 phosphorylation and elastin deposition were evaluated in lung tissue. Results: Total TGFβ1 and TGFβ2 decreased by 24 and 51% after combined UP+LPS exposure, whereas total TGFβ1 increased by 31% after 7 days' LPS exposure but not after double exposures. Alveolar expression of TGFβR2 decreased 75% after UP, but remained unaltered after double exposures. Decreased focal elastin deposition after single LPS exposure was prevented by double exposures. Conclusions: TGFβ signaling components and elastin responded differently to intrauterine LPS and UP exposure. Multiple bacterial exposures attenuated TGFβ signaling and normalized elastin deposition.

Functional analysis of missense variants in the TRESK (KCNK18) K+ channel

A loss of function mutation in the TRESK K2P potassium channel (KCNK18), has recently been linked with typical familial migraine with aura. We now report the functional characterisation of additional TRESK channel missense variants identified in unrelated patients. Several variants either had no apparent functional effect, or they caused a reduction in channel activity. However, the C110R variant was found to cause a complete loss of TRESK function, yet is present in both sporadic migraine and control cohorts, and no variation in KCNK18 copy number was found. Thus despite the previously identified association between loss of TRESK channel activity and migraine in a large multigenerational pedigree, this finding indicates that a single non-functional TRESK variant is not alone sufficient to cause typical migraine and highlights the genetic complexity of this disorder. Migraine is a common, disabling neurological disorder with a genetic, environmental and in some cases hormonal component. It is characterized by attacks of severe, usually unilateral and throbbing headache, can be accompanied by nausea, vomiting and photophobia and is clinically divided into two main subtypes, migraine with aura (MA) when a migraine is accompanied by transient and reversible focal neurological symptoms and migraine without aura (MO)1. The multifactorial and clinical heterogeneity of the disorder have considerably hindered the identification of common migraine susceptibility genes and most of our current understanding comes from the studies of familial hemiplegic migraine (FHM), a rare monogenic autosomal dominant form of MA2. So far, the three susceptibility genes that have been convincingly identified in FHM families all encode ion channels or transporters: CACNA1A encoding the α1 subunit of the Cav2.1 calcium channel3, SCN1A encoding the Nav1.1 sodium channel4 and ATP1A2 encoding the α2 subunit of the Na+/K+ pump5. It is believed that mutations in these genes may lead to increased efflux of glutamate and potassium in the synapse and thereby cause migraine by rendering the brain more susceptible to cortical spreading depression (CSD)6 which is thought to play a role in initiating a migraine attack7,8. However, these genes have not to date been implicated in common forms of migraine9. Nevertheless, current opinion suggests that typical migraine, like FHM, is also disorder of neuronal excitability, ion homeostasis and neurotransmitter release10,11,12. Mutations in the SLC4A4 gene encoding the sodium-bicarbonate cotransporter NBCe1, have recently been implicated in several different forms of migraine13, and a variety of genes involved in glutamate homeostasis (PGCP, MTDH14 and LRP115) and a cation channel (TRPM8)15 have also recently been implicated in migraine via genome-wide association studies. Ion channels are therefore highly likely to play an important role in the pathogenesis of typical migraine. TRESK (KCNK18), is a member of the two-pore domain (K2P) family of potassium channels involved in the control of cellular electrical excitability16. Regulation of TRESK activity by the calcium-dependent phosphatase calcineurin17, as well as its expression in dorsal root ganglia (DRG)18 and trigeminal ganglia (TG)19,20 has led to a proposed role for this channel in a variety of pain pathways. In a recent study, a frameshift mutation (F139Wfsx24) in TRESK was identified in a large multigenerational pedigree where it co-segregated perfectly with typical MA and a significant genome-wide linkage LOD score of 3.0. Furthermore, functional analysis revealed that this mutation caused a complete loss of TRESK function and that the truncated subunit was also capable of down regulating wild-type channel function. This therefore highlighted KCNK18 as potentially important candidate gene and suggested that TRESK dysfunction might play a possible role in the pathogenesis of familial migraine with visual aura20. Additional screening for KCNK18 mutations in unrelated sporadic migraine and control cohorts also identified a number of other missense variants; R10G, A34V, C110R, S231P and A233V20. The A233V variant was found only in the control cohort, whilst A34V was identified in a single Australian migraine proband for which family samples were not available, but it was not detected in controls. By contrast, the R10G, C110R, and S231P variants were found in both migraineurs and controls in both cohorts. In this study, we have investigated the functional effect of these variants to further probe the potential association of TRESK dysfunction with typical migraine.