Selective PEGylation of Parylene-C/SiO2 Substrates for Improved Astrocyte Cell Patterning.

Controlling the spatial distribution of glia and neurons in in vitro culture offers the opportunity to study how cellular interactions contribute to large scale network behaviour. A recently developed approach to cell-patterning uses differential adsorption of animal-serum protein on parylene-C and SiO2 surfaces to enable patterning of neurons and glia. Serum, however, is typically poorly defined and generates reproducibility challenges. Alternative activation methods are highly desirable to enable patterning without relying on animal serum. We take advantage of the innate contrasting surface chemistries of parylene-C and SiO2 to enable selective bonding of polyethylene glycol SiO2 surfaces, i.e. PEGylation, rendering them almost completely repulsive to cell adhesion. As the reagents used in the PEGylation protocol are chemically defined, the reproducibility and batch-to-batch variability complications associated with the used of animal serum are avoided. We report that PEGylated parylene-C/SiO2 substrates achieve a contrast in astrocyte density of 65:1 whereas the standard serum-immersion protocol results in a contrast of 5.6:1. Furthermore, single-cell isolation was significantly improved on PEGylated substrates when astrocytes were grown on close-proximity parylene-C nodes, whereas isolation was limited on serum-activated substrates due tolerance for cell adhesion on serum-adsorbed SiO2 surfaces.

Patterning of functional human astrocytes onto parylene-C/SiO2 substrates for the study of Ca2+ dynamics in astrocytic networks.

OBJECTIVE: Recent literature suggests that astrocytes form organized functional networks and communicate through transient changes in cytosolic Ca2+. Traditional techniques to investigate network activity, such as pharmacological blocking or genetic knockout, are difficult to restrict to individual cells. The objective of this work is to develop cell-patterning techniques to physically manipulate astrocytic interactions to enable the study of Ca2+ in astrocytic networks. APPROACH: We investigate how an in vitro cell-patterning platform that utilizes geometric patterns of parylene-C on SiO2 can be used to physically isolate single astrocytes and small astrocytic networks. MAIN RESULTS: We report that single astrocytes are effectively isolated on 75 × 75 µm square parylene nodes, whereas multi-cellular astrocytic networks are isolated on larger nodes, with the mean number of astrocytes per cluster increasing as a function of node size. Additionally, we report that astrocytes in small multi-cellular clusters exhibit spatio-temporal clustering of Ca2+ transients. Finally, we report that the frequency and regularity of Ca2+ transients was positively correlated with astrocyte connectivity. SIGNIFICANCE: The significance of this work is to demonstrate how patterning hNT astrocytes replicates spatio-temporal clustering of Ca2+ signalling that is observed in vivo but not in dissociated in vitro cultures. We therefore highlight the importance of the structure of astrocytic networks in determining ensemble Ca2+ behaviour.

Hazard perception skills of young drivers with Attention Deficit Hyperactivity Disorder (ADHD) can be improved with computer based driver training: An exploratory randomised controlled trial.

BACKGROUND: Young drivers with Attention Deficit Hyperactivity Disorder (ADHD) are at higher risk of road traffic injuries than their peers. Increased risk correlates with poor hazard perception skill. Few studies have investigated hazard perception training using computer technology with this group of drivers. OBJECTIVES: *Determine the presence and magnitude of the between-group and within- subject change in hazard perception skills in young drivers with ADHD who receive Drive Smart training. *Determine whether training-facilitated change in hazard perception is maintained over time. METHODS: This was a feasibility study, randomised control trial conducted in Australia. The design included a delayed treatment for the control group. Twenty-five drivers with a diagnosis of ADHD were randomised to the Immediate Intervention or Delayed Intervention group.The Immediate Intervention group received a training session using a computer application entitled Drive Smart. The Delayed Intervention group watched a documentary video initially (control condition), followed by the Drive Smart computer training session. The participant's hazard perception skill was measured using the Hazard Perception Test (HPT). FINDINGS: After adjusting for baseline scores, there was a significant betweengroup difference in post-intervention HPT change scores in favour of the Immediate Intervention group. The magnitude of the effect was large. There was no significant within-group delayed intervention effect. A significant maintenance effect was found at 6-week follow-up for the Immediate Intervention group. CONCLUSIONS: The hazard perception skills of participants improved following training with large effect size and some maintenance of gain. A multimodal approach to training is indicated to facilitate maintenance. A full-scale trial is feasible.

Nuclear Force Imprints Revealed on the Elastic Scattering of Protons with

How does nature hold together protons and neutrons to form the wide variety of complex nuclei in the Universe? Describing many-nucleon systems from the fundamental theory of quantum chromodynamics has been the greatest challenge in answering this question. The chiral effective field theory description of the nuclear force now makes this possible but requires certain parameters that are not uniquely determined. Defining the nuclear force needs identification of observables sensitive to the different parametrizations. From a measurement of proton elastic scattering on ^{10}C at TRIUMF and ab initio nuclear reaction calculations, we show that the shape and magnitude of the measured differential cross section is strongly sensitive to the nuclear force prescription.

Scattering of the Halo Nucleus

Angular distributions of the elastic, inelastic, and breakup cross sections of the halo nucleus ^{11}Be on ^{197}Au were measured at energies below (E_{lab}=31.9 MeV) and around (39.6 MeV) the Coulomb barrier. These three channels were unambiguously separated for the first time for reactions of ^{11}Be on a high-Z target at low energies. The experiment was performed at TRIUMF (Vancouver, Canada). The differential cross sections were compared with three different calculations: semiclassical, inert-core continuum-coupled-channels and continuum-coupled-channels ones with including core deformation. These results show conclusively that the elastic and inelastic differential cross sections can only be accounted for if core-excited admixtures are taken into account. The cross sections for these channels strongly depend on the B(E1) distribution in ^{11}Be, and the reaction mechanism is sensitive to the entanglement of core and halo degrees of freedom in ^{11}Be.

Nanosecond UV lasers stimulate transient Ca2+ elevations in human hNT astrocytes.

OBJECTIVE: Astrocytes respond to various stimuli resulting in intracellular Ca2+ signals that can propagate through organized functional networks. Recent literature calls for the development of techniques that can stimulate astrocytes in a fast and highly localized manner to emulate more closely the characteristics of astrocytic Ca2+ signals in vivo. APPROACH: In this article we demonstrate, for the first time, how nanosecond UV lasers are capable of reproducibly stimulating Ca2+ transients in human hNT astrocytes. MAIN RESULTS: We report that laser pulses with a beam energy of 4-29 µJ generate transient increases in cytosolic Ca2+. These Ca2+ transients then propagate to adjacent astrocytes as intercellular Ca2+ waves. SIGNIFICANCE: We propose that nanosecond laser stimulation provides a valuable tool for enabling the study of Ca2+ dynamics in human astrocytes at both a single cell and network level. Compared to previously developed techniques nanosecond laser stimulation has the advantage of not requiring loading of photo-caged or -sensitising agents, is non-contact, enables stimulation with a high spatiotemporal resolution and is comparatively cost effective.

Longitudinal employment outcomes of an early intervention vocational rehabilitation service for people admitted to rehabilitation with a traumatic spinal cord injury.

STUDY DESIGN: Longitudinal cohort design. OBJECTIVES: First, to explore the longitudinal outcomes for people who received early intervention vocational rehabilitation (EIVR); second, to examine the nature and extent of relationships between contextual factors and employment outcomes over time. SETTING: Both inpatient and community-based clients of a Spinal Community Integration Service (SCIS). METHODS: People of workforce age undergoing inpatient rehabilitation for traumatic spinal cord injury were invited to participate in EIVR as part of SCIS. Data were collected at the following three time points: discharge and at 1 year and 2+ years post discharge. Measures included the spinal cord independence measure, hospital anxiety and depression scale, impact on participation and autonomy scale, numerical pain-rating scale and personal wellbeing index. A range of chi square, correlation and regression tests were undertaken to look for relationships between employment outcomes and demographic, emotional and physical characteristics. RESULTS: Ninety-seven participants were recruited and 60 were available at the final time point where 33% (95% confidence interval (CI): 24-42%) had achieved an employment outcome. Greater social participation was strongly correlated with wellbeing (ρ=0.692), and reduced anxiety (ρ=-0.522), depression (ρ=-0.643) and pain (ρ=-0.427) at the final time point. In a generalised linear mixed effect model, education status, relationship status and subjective wellbeing increased significantly the odds of being employed at the final time point. Tertiary education prior to injury was associated with eight times increased odds of being in employment at the final time point; being in a relationship at the time of injury was associated with increased odds of being in employment of more than 3.5; subjective wellbeing, while being the least powerful predictor was still associated with increased odds (1.8 times) of being employed at the final time point. CONCLUSIONS: EIVR shows promise in delivering similar return-to-work rates as those traditionally reported, but sooner. The dynamics around relationships, subjective wellbeing, social participation and employment outcomes require further exploration.

Human astrocytic grid networks patterned in parylene-C inlayed SiO2 trenches.

Recent literature suggests that glia, and in particular astrocytes, should be studied as organised networks which communicate through gap junctions. Astrocytes, however, adhere to most surfaces and are highly mobile cells. In order to study, such organised networks effectively in vitro it is necessary to influence them to pattern to certain substrates whilst being repelled from others and to immobilise the astrocytes sufficiently such that they do not continue to migrate further whilst under study. In this article, we demonstrate for the first time how it is possible to facilitate the study of organised patterned human astrocytic networks using hNT astrocytes in a SiO2 trench grid network that is inlayed with the biocompatible material, parylene-C. We demonstrate how the immobilisation of astrocytes lies in the depth of the SiO2 trench, determining an optimum trench depth and that the optimum patterning of astrocytes is a consequence of the parylene-C inlay and the grid node spacing. We demonstrate high fidelity of the astrocytic networks and demonstrate that functionality of the hNT astrocytes through ATP evoked calcium signalling is also dependent on the grid node spacing. Finally, we demonstrate that the location of the nuclei on the grid nodes is also a function of the grid node spacing. The significance of this work, is to describe a suitable platform to facilitate the study of hNT astrocytes from the single cell level to the network level to improve knowledge and understanding of how communication links to spatial organisation at these higher order scales and trigger in vitro research further in this area with clinical applications in the area of epilepsy, stroke and focal cerebral ischemia.

High-Precision Half-Life Measurements for the Superallowed ß

Precision measurements of superallowed Fermi ß-decay transitions, particularly for the lightest superallowed emitters ^{10}C and ^{14}O, set stringent limits on possible scalar current contributions to the weak interaction. In the present work, a discrepancy between recent measurements of the ^{10}C half-life is addressed through two high-precision half-life measurements, via γ-ray photopeak and ß counting, that yield consistent results for the ^{10}C half-life of T_{1/2}=19.2969±0.0074 s and T_{1/2}=19.3009±0.0017 s, respectively. The latter is the most precise superallowed ß-decay half-life measurement reported to date and the first to achieve a relative precision below 10^{-4}. A fit to the world superallowed ß-decay data including the ^{10}C half-life measurements reported here yields b_{F}=-0.0018±0.0021 (68% C.L.) for the Fierz interference term and C_{S}/C_{V}=+0.0009±0.0011 for the ratio of the weak scalar to vector couplings assuming left-handed neutrinos.

Evidence of soft dipole resonance in

The first conclusive evidence of a dipole resonance in ^{11}Li having isoscalar character observed from inelastic scattering with a novel solid deuteron target is reported. The experiment was performed at the newly commissioned IRIS facility at TRIUMF. The results show a resonance peak at an excitation energy of 1.03±0.03 MeV with a width of 0.51±0.11 MeV (FWHM). The angular distribution is consistent with a dipole excitation in the distorted-wave Born approximation framework. The observed resonance energy together with shell model calculations show the first signature that the monopole tensor interaction is important in ^{11}Li. The first ab initio calculations in the coupled cluster framework are also presented.

Low cost, patterning of human hNT brain cells on parylene-C with UV & IR laser machining.

This paper describes the use of 800nm femtosecond infrared (IR) and 248nm nanosecond ultraviolet (UV) laser radiation in performing ablative micromachining of parylene-C on SiO2 substrates for the patterning of human hNT astrocytes. Results are presented that support the validity of using IR laser ablative micromachining for patterning human hNT astrocytes cells while UV laser radiation produces photo-oxidation of the parylene-C and destroys cell patterning. The findings demonstrate how IR laser ablative micromachining of parylene-C on SiO2 substrates can offer a low cost, accessible alternative for rapid prototyping, high yield cell patterning.

Infra-red laser ablative micromachining of parylene-C on SiO2 substrates for rapid prototyping, high yield, human neuronal cell patterning.

Cell patterning commonly employs photolithographic methods for the micro fabrication of structures on silicon chips. These require expensive photo-mask development and complex photolithographic processing. Laser based patterning of cells has been studied in vitro and laser ablation of polymers is an active area of research promising high aspect ratios. This paper disseminates how 800 nm femtosecond infrared (IR) laser radiation can be successfully used to perform laser ablative micromachining of parylene-C on SiO2 substrates for the patterning of human hNT astrocytes (derived from the human teratocarcinoma cell line (hNT)) whilst 248 nm nanosecond ultra-violet laser radiation produces photo-oxidization of the parylene-C and destroys cell patterning. In this work, we report the laser ablation methods used and the ablation characteristics of parylene-C for IR pulse fluences. Results follow that support the validity of using IR laser ablative micromachining for patterning human hNT astrocytes cells. We disseminate the variation in yield of patterned hNT astrocytes on parylene-C with laser pulse spacing, pulse number, pulse fluence and parylene-C strip width. The findings demonstrate how laser ablative micromachining of parylene-C on SiO2 substrates can offer an accessible alternative for rapid prototyping, high yield cell patterning with broad application to multi-electrode arrays, cellular micro-arrays and microfluidics.

First human hNT neurons patterned on parylene-C/silicon dioxide substrates: Combining an accessible cell line and robust patterning technology for the study of the pathological adult human brain.

In this communication, we describe a new method which has enabled the first patterning of human neurons (derived from the human teratocarcinoma cell line (hNT)) on parylene-C/silicon dioxide substrates. We reveal the details of the nanofabrication processes, cell differentiation and culturing protocols necessary to successfully pattern hNT neurons which are each key aspects of this new method. The benefits in patterning human neurons on silicon chip using an accessible cell line and robust patterning technology are of widespread value. Thus, using a combined technology such as this will facilitate the detailed study of the pathological human brain at both the single cell and network level.

Excessive noise injection training of neural networks for markerless tracking in obscured and segmented environments.

In this letter, we demonstrate that the generalization properties of a neural network (NN) can be extended to encompass objects that obscure or segment the original image in its foreground or background. We achieve this by piloting an extension of the noise injection training technique, which we term excessive noise injection (ENI), on a simple feedforward multilayer perceptron (MLP) network with vanilla backward error propagation to achieve this aim. Six tests are reported that show the ability of an NN to distinguish six similar states of motion of a simplified human figure that has become obscured by moving vertical and horizontal bars and random blocks for different levels of obscuration. Four more extensive tests are then reported to determine the bounds of the technique. The results from the ENI network were compared to results from the same NN trained on clean states only. The results pilot strong evidence that it is possible to track a human subject behind objects using this technique, and thus this technique lends itself to a real-time markerless tracking system from a single video stream.

Selection for rehabilitation: acute care discharge patterns for stroke and orthopaedic patients.

This study aimed to develop predictive models to assist clinicians working in acute care to determine which stroke and lower limb orthopaedic patients should be discharged to nursing homes, inpatient rehabilitation, or home. The subjects were 223 patients with stroke and 125 patients with lower limb orthopaedic problems, aged 60 years and over, and the 22 acute care and rehabilitation clinicians who cared for these patients. Patient status was measured within 72 hours of discharge using the Adult FIM(SM) (Guide for the Uniform Data Set for Medical Rehabilitation, 1993) and ten additional items from either the RICFAS (Rehabilitation Institute of Chicago, 1987) or developed for this research. Four mathematical models were developed to predict discharge destination using stepwise discriminant function analysis. Using between three and seven items from the FIM(SM) alone or FIM(SM) plus additional variables, these models correctly classified between 74.9% and 80.5% of patients. In conclusion, clinicians are offered a selection of models to guide their discharge destination decisions for two large groups of patients. While these models cannot replace clinical judgement, they increase the transparency of decisions. The use of objective models to guide clinical decisions are essential, particularly given increasing pressure to justify patient access to costly rehabilitation services.

Using a head-mounted video camera to study clinical reasoning.

Studies of clinical reasoning are essential if we are to extend our knowledge of occupational therapy practice, better communicate our work to clients and colleagues, and reveal to our students the nuances of therapy that cannot be gained from texts. However, accessing therapists' clinical reasoning is not an easy task because these cognitive processes can be studied only indirectly. The aim of this article is to promote the study of clinical reasoning by reviewing a new approach to data collection in this field. To achieve this purpose, the article outlines current data collection methods, such as the think-aloud method written notes, free recall, and audio-assisted and video-assisted recall. A novel method involving a head-mounted video camera is described, and details are provided on a modified approach to debriefing using video-assisted recall. Anecdotes from the authors experience of using this technology illustrate the text.

Development of an assessment to identify play behaviours that discriminate between the play of typical preschoolers and preschoolers with pre-academic problems.

Occupational therapists regard play as the primary occupation of childhood. However, many authors of play assessments have viewed play either as a functional outcome of other skills or as an indication of the child's developmental level. While such views of play are valid, these approaches to play are broad and do not fully consider the cognitive value of play. A study was undertaken with 82 preschoolers to see if a new play assessment which considers cognitive play skills, called the Child-initiated Pretend Play Assessment, (ChiPPA), possessed acceptable inter-rater reliability, and could discriminate between the play of typically developing preschoolers and preschoolers with pre-academic problems. This assessment evaluates preschoolers' elaborate imaginative actions, number of imitative actions, and number of object substitutions. The ChiPPA was found to have excellent inter-rater reliability and did discriminate between children who were typically developing and children who experienced pre-academic problems. In particular, the elaborateness of a child's play actions and the child's ability to substitute objects were strong discriminators of preschoolers with and without pre-academic problems. Three play profiles which emerged from the research are discussed.

Psychological sequelae of torture and organised violence suffered by refugees from Iraq. Trauma-related factors compared with social factors in exile.

BACKGROUND: Refugees who have suffered traumatic events present complex therapeutic challenges to health professionals. There is little research into post-exile factors that may be amenable to change, and therefore reduce morbidity. We examined the importance of social factors in exile and of trauma factors in producing the different elements of psychological sequelae of severe trauma. METHOD: Eighty-four male Iraqi refugees were interviewed. Adverse events and level of social support were measured. Various measures of psychological morbidity were applied, all of which have been used in previous trauma research. RESULTS: Social factors in exile, particularly the level of "affective" social support, proved important in determining the severity of both post-traumatic stress disorder and depressive reactions, particularly when combined with a severe level of trauma/torture. Poor social support is a stronger predictor of depressive morbidity than trauma factors. CONCLUSIONS: Some of the most important factors in producing psychological morbidity in refugees may be alleviated by planned, integrated rehabilitation programmes and attention to social support and family reunion.

Trauma-rehabilitation connections: discharge and admission decisions for children.

Paediatric discharge coordinators and rehabilitation admission coordinators were surveyed about how children are selected for rehabilitation. The following areas are covered: (1) the decision process, and how children are selected for rehabilitation; (2) who is involved in making acute-care discharge and rehabilitation admission decisions; (3) factors that guide selection of children for rehabilitation; and (4) satisfaction with referral practices. Surveys were completed by 30 trauma discharge and rehabilitation admission coordinators, recruited from paediatric trauma units and paediatric/general rehabilitation units nationwide. Most respondents were satisfied with transfers, although some voiced concerns about constraints placed on referrals by insurance. Even when inpatient rehabilitation was clearly needed, 40% said insurance status still affected whether children were admitted. There was little evidence that any uniform criteria are used to make decisions. Half had no training in discharge/admission planning and half did not base decisions on functional assessments. Although guidelines are increasingly used in clinical decision-making, few are available concerning critical decisions about which children receive inpatient rehabilitation following trauma.