Understanding the abnormal brain activity in epilepsy as a potential predictor of the onset of an epileptic seizure

The Brain Research Institute (BRI) uses various types of indirect measurements, including EEG and fMRI, to understand and assess brain activity and function. As well as the recovery of generic information about brain function, research also focuses on the utilisation of such data and understanding to study the initiation, dynamics, spread and suppression of epileptic seizures. To assist with the future focussing of this aspect of their research, the BRI asked the MISG 2010 participants to examine how the available EEG and fMRI data and current knowledge about epilepsy should be analysed and interpreted to yield an enhanced understanding about brain activity occurring before, at commencement of, during, and after a seizure. Though the deliberations of the study group were wide ranging in terms of the related matters considered and discussed, considerable progress was made with the following three aspects. (1) The science behind brain activity investigations depends crucially on the quality of the analysis and interpretation of, as well as the recovery of information from, EEG and fMRI measurements. A number of specific methodologies were discussed and formalised, including independent component analysis, principal component analysis, profile monitoring and change point analysis (hidden Markov modelling, time series analysis, discontinuity identification). (2) Even though EEG measurements accurately and very sensitively record the onset of an epileptic event or seizure, they are, from the perspective of understanding the internal initiation and localisation, of limited utility. They only record neuronal activity in the cortical (surface layer) neurons of the brain, which is a direct reflection of the type of electrical activity they have been designed to record. Because fMRI records, through the monitoring of blood flow activity, the location of localised brain activity within the brain, the possibility of combining fMRI measurements with EEG, as a joint inversion activity, was discussed and examined in detail. (3) A major goal for the BRI is to improve understanding about ``when'' (at what time) an epileptic seizure actually commenced before it is identified on an eeg recording, ``where'' the source of this initiation is located in the brain, and ``what'' is the initiator. Because of the general agreement in the literature that, in one way or another, epileptic events and seizures represent abnormal synchronisations of localised and/or global brain activity the modelling of synchronisations was examined in some detail. References C. M. Michel, G. Thut, S. Morand, A. Khateb, A. J. Pegna, R. Grave de Peralta, S. Gonzalez, M. Seeck and T. Landis, Electric source imaging of human brain functions, Brain Res. 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Genomic organisation, activity and distribution analysis of the microbial putrescine oxidase degradation pathway

The catalytic action of putrescine specific amine oxidases acting in tandem with 4-aminobutyraldehyde dehydrogenase is explored as a degradative pathway in Rhodococcus opacus. By limiting the nitrogen source, increased catalytic activity was induced leading to a coordinated response in the oxidative deamination of putrescine to 4-aminobutyraldehyde and subsequent dehydrogenation to 4-aminobutyrate. Isolating the dehydrogenase by ion exchange chromatography and gel filtration revealed that the enzyme acts principally on linear aliphatic aldehydes possessing an amino moiety. Michaelis-Menten kinetic analysis delivered a Michaelis constant (KM=0.014mM) and maximum rate (Vmax=11.2μmol/min/mg) for the conversion of 4-aminobutyraldehyde to 4-aminobutyrate. The dehydrogenase identified by MALDI-TOF mass spectrometric analysis (E value=0.031, 23% coverage) belongs to a functionally related genomic cluster that includes the amine oxidase, suggesting their association in a directed cell response. Key regulatory, stress and transport encoding genes have been identified, along with candidate dehydrogenases and transaminases for the further conversion of 4-aminobutyrate to succinate. Genomic analysis has revealed highly similar metabolic gene clustering among members of Actinobacteria, providing insight into putrescine degradation notably among Micrococcaceae, Rhodococci and Corynebacterium by a pathway that was previously uncharacterised in bacteria.

Effects of dissolved oxygen availability and culture biomass at induction upon the intracellular expression of monoamine oxidase by recombinant E. coli in fed batch bioprocesses

The effects of oxygen availability and induction culture biomass upon production of an industrially important monoamine oxidase (MAO) were investigated in fed-batch cultures of a recombinant E. coli. For each induction cell biomass 2 different oxygenation methods were used, aeration and oxygen enriched air. Induction at higher biomass levels increased the culture demand for oxygen, leading to fermentative metabolism and accumulation of high levels of acetate in the aerated cultures. Paradoxically, despite an almost eight fold increase in acetate accumulation to levels widely reported to be highly detrimental to protein production, when induction wet cell weight (WCW) rose from 100% to 137.5%, MAO specific activity in these aerated processes showed a 3 fold increase. By contrast, for oxygenated cultures induced at WCW's 100% and 137.5% specific activity levels were broadly similar, but fell rapidly after the maxima were reached. Induction at high biomass levels (WCW 175%) led to very low levels of specific MAO activity relative to induction at lower WCW's in both aerated and oxygenated cultures. Oxygen enrichment of these cultures was a useful strategy for boosting specific growth rates, but did not have positive effects upon specific enzyme activity. Based upon our findings, consideration of the amino acid composition of MAO and previous studies on related enzymes, we propose that this effect is due to oxidative damage to the MAO enzyme itself during these highly aerobic processes. Thus, the optimal process for MAO production is aerated, not oxygenated, and induced at moderate cell density, and clearly represents a compromise between oxygen supply effects on specific growth rate/induction cell density, acetate accumulation, and high specific MAO activity. This work shows that the negative effects of oxygen previously reported in free enzyme preparations, are not limited to these acellular environments but are also discernible in the sheltered environment of the cytosol of E. coli cells.

Racial differences in physical activity among breast cancer survivors : implications for breast cancer care

Background: Physical activity after breast cancer diagnosis is associated with improved survival. This study examines levels of and changes in physical activity following breast cancer diagnosis, overall and by race. Methods: The Carolina Breast Cancer Study, Phase III, assessed pre- and post-diagnosis physical activity levels in a cohort of 1,735 women, aged 20-74, diagnosed with invasive breast cancer between 2008 and 2011 in 44 counties of North Carolina. Logistic regression and analysis of variance were used to examine whether demographic, behavioral and clinical characteristics were associated with activity levels. Results: Only 35% of breast cancer survivors met current physical activity guidelines post-diagnosis. A decrease in activity following diagnosis was reported by 59% of patients, with the average study participant reducing their activity by 230 minutes (95% CI: 190, 270). Following adjustment for potential confounders, when compared to white women, African-American women were less likely to meet national physical activity guidelines post-diagnosis (odds ratio: 1.38, 95% CI: 1.01, 1.88), reported less weekly post-diagnosis physical activity (182 vs. 215 minutes; p=0.13), and reported higher average reductions in pre- versus post-diagnosis weekly activity (262 vs. 230 minutes; p-value = 0.13). Conclusion: Despite compelling evidence demonstrating the benefits of physical activity post-breast cancer, it is clear that more work needs to be done to promote physical activity in breast cancer patients, especially among African-American women.

Bisphenol A degradation enhanced by air bubbles via advanced oxidation using in situ generated ferrous ions from nano zero-valent iron/palygorskite composite materials

Novel nano zero-valent iron/palygorskite composite materials prepared by evaporative and centrifuge methods are tested for the degradation of bisphenol A in an aqueous medium. A systematic study is presented which showed that nano zero-valent iron material has little effect on bisphenol A degradation. When hydrogen peroxide was added to initiate the reaction, some percentage of bisphenol A removal (∼20%) was achieved; however, with the aid of air bubbles, the percentage removal can be significantly increased to ∼99%. Compared with pristine nano zero-valent iron and commercial iron powder, nano zero-valent iron/palygorskite composite materials have much higher reactivity towards bisphenol A and these materials are superior as they have little impact on the solution pH. However, for pristine nano zero-valent iron, it is difficult to maintain the reaction system at a favourable low pH which is a key factor in maintaining high bisphenol A removal. All materials were characterized by X-ray diffraction, scanning electron microscopy, elemental analysis, transmission electron microscopy and X-ray photoelectron spectroscopy. The optimum conditions were obtained based on a series of batch experiments. This study has extended the application of nano zero-valent iron/palygorskite composites as effective materials for the removal of phenolic compounds from the environment.

Investigation of air quality at Lithgow Correctional Centre

This project was conducted at Lithgow Correctional Centre (LCC), NSW, Australia. Air quality field measurements were conducted on two occasions (23-27 May 2012, and 3-8 December 2012), just before and six months after the introduction of smoke free buildings policies (28 May 2012) at the LCC, respectively. The main aims of this project were to: (1) investigate the indoor air quality; (2) quantify the level of exposure to environmental tobacco smoke (ETS); (3) identify the main indoor particle sources; (4) distinguish between PM2.5 / particle number from ETS, as opposed to other sources; and (5) provide recommendations for improving indoor air quality and/or minimising exposure at the LCC. The measurements were conducted in Unit 5.2A, Unit 5.2B, Unit 1.1 and Unit 3.1, together with personal exposure measurements, based on the following parameters: -Indoor and outdoor particle number (PN) concentration in the size range 0.005-3 µm -Indoor and outdoor PM2.5 particle mass concentration -Indoor and outdoor VOC concentrations -Personal particle number exposure levels (in the size range 0.01-0.3 µm) -Indoor and outdoor CO and CO2 concentrations, temperature and relative humidity In order to enhance the outcomes of this project, the indoor and outdoor particle number (PN) concentrations were measured by two additional instruments (CPC 3787) which were not listed in the original proposal.

Aligning in and through interaction : children getting in and out of spontaneous activity

Spontaneous play, important for forming the basis of friendships and peer relations, is a complex activity involving the management and production of talk-in-interaction. This paper focuses on the intricacies of social interaction, emphasising the link between alignment and affiliation, and the range and importance of verbal and nonverbal interactive devices available to children. Analysis of the way in which two girls, one of whom has been diagnosed with Asperger’s Syndrome, engage in spontaneous activities demonstrates the potential for interactional difficulty due to the unscripted nature of the interaction. The paper argues for further research into how improvised, unscripted interactions are initiated within moment-by-moment talk, how they unfold, and how they are brought to a close in everyday contexts in order to understand how children create their social worlds.

The effect of very small air gaps on small field dosimetry

The purpose of this study was to investigate the effect of very small air gaps (less than 1 mm) on the dosimetry of small photon fields used for stereotactic treatments. Measurements were performed with optically stimulated luminescent dosimeters (OSLDs) for 6 MV photons on a Varian 21iX linear accelerator with a Brainlab lMLC attachment for square field sizes down to 6 mm 9 6 mm. Monte Carlo simulations were performed using EGSnrc C++ user code cavity. It was found that the Monte Carlo model used in this study accurately simulated the OSLD measurements on the linear accelerator. For the 6 mm field size, the 0.5 mm air gap upstream to the active area of the OSLD caused a 5.3 % dose reduction relative to a Monte Carlo simulation with no air gap...

Red colouration in apple fruit is due to the activity of the MYB transcription factor, MdMYB10

Anthocyanin concentration is an important determinant of the colour of many fruits. In apple (Malus x domestica), centuries of breeding have produced numerous varieties in which levels of anthocyanin pigment vary widely and change in response to environmental and developmental stimuli. The apple fruit cortex is usually colourless, although germplasm does exist where the cortex is highly pigmented due to the accumulation of either anthocyanins or carotenoids. From studies in a diverse array of plant species, it is apparent that anthocyanin biosynthesis is controlled at the level of transcription. Here we report the transcript levels of the anthocyanin biosynthetic genes in a red-fleshed apple compared with a white-fleshed cultivar. We also describe an apple MYB transcription factor, MdMYB10, that is similar in sequence to known anthocyanin regulators in other species. We further show that this transcription factor can induce anthocyanin accumulation in both heterologous and homologous systems, generating pigmented patches in transient assays in tobacco leaves and highly pigmented apple plants following stable transformation with constitutively expressed MdMYB10. Efficient induction of anthocyanin biosynthesis in transient assays by MdMYB10 was dependent on the co-expression of two distinct bHLH proteins from apple, MdbHLH3 and MdbHLH33. The strong correlation between the expression of MdMYB10 and apple anthocyanin levels during fruit development suggests that this transcription factor is responsible for controlling anthocyanin biosynthesis in apple fruit; in the red-fleshed cultivar and in the skin of other varieties, there is an induction of MdMYB10 expression concurrent with colour formation during development. Characterization of MdMYB10 has implications for the development of new varieties through classical breeding or a biotechnological approach.

Transient expression vectors for functional genomics, quantification of promoter activity and RNA silencing in plants

Background We describe novel plasmid vectors for transient gene expression using Agrobacterium, infiltrated into Nicotiana benthamiana leaves. We have generated a series of pGreenII cloning vectors that are ideally suited to transient gene expression, by removing elements of conventional binary vectors necessary for stable transformation such as transformation selection genes. Results We give an example of expression of heme-thiolate P450 to demonstrate effectiveness of this system. We have also designed vectors that take advantage of a dual luciferase assay system to analyse promoter sequences or post-transcriptional regulation of gene expression. We have demonstrated their utility by co-expression of putative transcription factors and the promoter sequence of potential target genes and show how orthologous promoter sequences respond to these genes. Finally, we have constructed a vector that has allowed us to investigate design features of hairpin constructs related to their ability to initiate RNA silencing, and have used these tools to study cis-regulatory effect of intron-containing gene constructs. Conclusion In developing a series of vectors ideally suited to transient expression analysis we have provided a resource that further advances the application of this technology. These minimal vectors are ideally suited to conventional cloning methods and we have used them to demonstrate their flexibility to investigate enzyme activity, transcription regulation and post-transcriptional regulatory processes in transient assays.

Age and gender differences in objectively measured physical activity in youth

Purpose The purpose of this study was to evaluate age and gender differences in objectively measured physical activity (PA) in a population-based sample of students in grades 1–12. Methods Participants (185 male, 190 female) wore a CSA 7164 accelerometer for 7 consecutive days. To examine age-related trends, students were grouped as follows: grades 1–3 (N = 90), grades 4–6 (N = 91), grades 7–9 (N = 96), and grades 10–12 (N = 92). Bouts of PA and minutes spent in moderate-to-vigorous PA (MVPA) and vigorous PA (VPA) were examined. Results Daily MVPA and VPA exhibited a significant inverse relationship with grade level, with the largest differences occurring between grades 1–3 and 4–6. Boys were more active than girls; however, for overall PA, the magnitudes of the gender differences were modest. Participation in continuous 20-min bouts of PA was low to nonexistent. Conclusion Our results support the notion that PA declines rapidly during childhood and adolescence and that accelerometers are feasible alternatives to self-report methods in moderately sized population-level surveillance studies.

Conducting accelerometer-based activity assessments in field-based research

Purpose The purpose of this review is to address important methodological issues related to conducting accelerometer-based assessments of physical activity in free-living individuals. Methods We review the extant scientific literature for empirical information related to the following issues: product selection, number of accelerometers needed, placement of accelerometers, epoch length, and days of monitoring required to estimate habitual physical activity. We also discuss the various options related to distributing and collecting monitors and strategies to enhance compliance with the monitoring protocol. Results No definitive evidence exists currently to indicate that one make and model of accelerometer is more valid and reliable than another. Selection of accelerometer therefore remains primarily an issue of practicality, technical support, and comparability with other studies. Studies employing multiple accelerometers to estimate energy expenditure report only marginal improvements in explanatory power. Accelerometers are best placed on hip or the lower back. Although the issue of epoch length has not been studied in adults, the use of count cut points based on 1-min time intervals maybe inappropriate in children and may result in underestimation of physical activity. Among adults, 3–5 d of monitoring is required to reliably estimate habitual physical activity. Among children and adolescents, the number of monitoring days required ranges from 4 to 9 d, making it difficult to draw a definitive conclusion for this population. Face-to-face distribution and collection of accelerometers is probably the best option in field-based research, but delivery and return by express carrier or registered mail is a viable option. Conclusion Accelerometer-based activity assessments requires careful planning and the use of appropriate strategies to increase compliance.

Correlates of adults' participation in physical activity : review and update

Purpose To review and update the evidence relating to the personal, social, and environmental factors associated with physical activity (PA) in adults. Methods Systematic review of the peer-reviewed literature to identify papers published between 1998 and 2000 with PA (and including exercise and exercise adherence). Qualitative reports or case studies were not included. Results Thirty-eight new studies were located. Most confirmed the existence of factors already known to be correlates of PA. Changes in status were noted in relation to the influence of marital status, obesity, smoking, lack of time, past exercise behavior, and eight environmental variables. New studies were located which focused on previously understudied population groups such as minorities, middle and older aged adults, and the disabled. Conclusion The newly reported studies tend to take a broader “ecological” approach to understanding the correlates of PA and are more focused on environmental factors. There remains a need to better understand environmental influences and the factors that influence different types of PA. As most of the work in this field still relies on cross-sectional studies, longitudinal and intervention studies will be required if causal relationships are to be inferred.

Evaluating a model of parental influence on youth physical activity

Objective To test a conceptual model linking parental physical activity orientations, parental support for physical activity, and children's self-efficacy perceptions with physical activity participation. Participants and setting The sample consisted of 380 students in grades 7 through 12 (mean age, 14.0±1.6 years) and their parents. Data collection took place during the fall of 1996. Main outcome measures Parents completed a questionnaire assessing their physical activity habits, enjoyment of physical activity, beliefs regarding the importance of physical activity, and supportive behaviors for their child's physical activity. Students completed a 46-item inventory assessing physical activity during the previous 7 days and a 5-item physical activity self-efficacy scale. The model was tested via observed variable path analysis using structural equation modeling techniques (AMOS 4.0). Results An initial model, in which parent physical activity orientations predicted child physical activity via parental support and child self-efficacy, did not provide an acceptable fit to the data. Inclusion of a direct path from parental support to child physical activity and deletion of a nonsignificant path from parental physical activity to child physical activity significantly improved model fit. Standardized path coefficients for the revised model ranged from 0.17 to 0.24, and all were significant at the p<0.0001 level. Conclusions Parental support was an important correlate of youth physical activity, acting directly or indirectly through its influence on self-efficacy. Physical activity interventions targeted at youth should include and evaluate the efficacy of individual-level and community-level strategies to increase parents’ capacity to provide instrumental and motivational support for their children's physical activity.

Evidence based physical activity for school-age youth

Objectives To review the effects of physical activity on health and behavior outcomes and develop evidence-based recommendations for physical activity in youth. Study design A systematic literature review identified 850 articles; additional papers were identified by the expert panelists. Articles in the identified outcome areas were reviewed, evaluated and summarized by an expert panelist. The strength of the evidence, conclusions, key issues, and gaps in the evidence were abstracted in a standardized format and presented and discussed by panelists and organizational representatives. Results Most intervention studies used supervised programs of moderate to vigorous physical activity of 30 to 45 minutes duration 3 to 5 days per week. The panel believed that a greater amount of physical activity would be necessary to achieve similar beneficial effects on health and behavioral outcomes in ordinary daily circumstances (typically intermittent and unsupervised activity). Conclusion School-age youth should participate daily in 60 minutes or more of moderate to vigorous physical activity that is developmentally appropriate, enjoyable, and involves a variety of activities.