Assaying protein palmitoylation in plants

<p>Abstract</p> <p>Background</p> <p>Protein S-acylation (also known as palmitoylation) is the reversible post-translational addition of acyl lipids to cysteine residues in proteins through a thioester bond. It allows strong association with membranes. Whilst prediction methods for S-acylation exist, prediction is imperfect. Existing protocols for demonstrating the S-acylation of plant proteins are either laborious and time consuming or expensive.</p> <p>Results</p> <p>We describe a biotin switch method for assaying the S-acylation of plant proteins. We demonstrate the technique by showing that the heterotrimeric G protein subunit AGG2 is S-acylated as predicted by mutagenesis experiments. We also show that a proportion of the Arabidopsis alpha-tubulin subunit pool is S-acylated <it>in planta</it>. This may account for the observed membrane association of plant microtubules. As alpha-tubulins are ubiquitously expressed they can potentially be used as a positive control for the S-acylation assay regardless of the cell type under study.</p> <p>Conclusion</p> <p>We provide a robust biotin switch protocol that allows the rapid assay of protein S-acylation state in plants, using standard laboratory techniques and without the need for expensive or specialised equipment. We propose alpha-tubulin as a useful positive control for the protocol.</p

Technology and its role in rehabilitation for people with cognitive-communication disability following a traumatic brain injury (TBI)

© 2017 Taylor & Francis Group, LLC. Purpose: To review the literature on communication technologies in rehabilitation for people with a traumatic brain injury (TBI), and: (a) determine its application to cognitive-communicative rehabilitation, and b) develop a model to guide communication technology use with people after TBI. Method: This integrative literature review of communication technology in TBI rehabilitation and cognitive-communication involved searching nine scientific databases and included 95 studies. Results: Three major types of communication technologies (assistive technology, augmentative and alternative communication technology, and information communication technology) and multiple factors relating to use of technology by or with people after TBI were categorized according to: (i) individual needs, motivations and goals; (ii) individual impairments, activities, participation and environmental factors; and (iii) technologies. While there is substantial research relating to communication technologies and cognitive rehabilitation after TBI, little relates specifically to cognitive-communication rehabilitation. Conclusions: Further investigation is needed into the experiences and views of people with TBI who use communication technologies, to provide the ‘user’ perspective and influence user-centred design. Research is necessary to investigate the training interventions that address factors fundamental for success, and any impact on communication. The proposed model provides an evidence-based framework for incorporating technology into speech pathology clinical practice and research

Content Analysis of Tweets by People with Traumatic Brain Injury (TBI): Implications for Rehabilitation and Social Media Goals

In this Twitter research, 6874 tweets of six adults with traumatic brain injury (TBI) were analyzed qualitatively and quantitatively using content classification [1], inductive coding of content themes, socio-linguistic analysis, and computational analysis in KH Coder. The results reflected that participants used Twitter for: (i) supporting others, including people with TBI; (ii) discussing society and culture, popular issues, news, and personal interests; (iii) connecting with others; (iv) sharing their experiences of life after TBI; (v) knowledge via exchanging information; and (vii) advocacy. ‘Emotional expression’, and ‘connection’ were common threads running across themes. Attending to the expressions of people with TBI on Twitter provides important insights into their lived experiences and could inform the development of user-centered cognitive-communication and social participation goals for people with TBI

Seasonality, phytoplankton succession and the biogeochemical impacts of an autumn storm in the northeast Atlantic Ocean

Phytoplankton chemotaxonomic distributions are examined in conjunction with taxon specific particulate biomass concentrations and phytoplankton abundances to investigate the biogeochemical consequences of the passage of an autumn storm in the northeast Atlantic Ocean. Chemotaxonomy indicated that the phytoplankton community was dominated by nanoplankton (2-20 μ), which on average represented 75±8% of the community. Microplankton (20-200 μ) and picoplankton (<2 μ) represented 21±7% and 4±3% respectively with the microplankton group composed of almost equal proportions of diatoms (53±17%) and dinoflagellates (47±17%). Total chlorophyll-a (TCHLa = CHLa + Divinyl CHLa) concentrations ranged from 22 to 677 ng L-1, with DvCHLa making minor contributions of between <1% and 13% to TCHLa. Higher DvCHLa contributions were seen during the storm, which deepened the surface mixed layer, increased mixed layer nutrient concentrations and vertically mixed the phytoplankton community leading to a post-storm increase in surface chlorophyll concentrations. Picoplankton were rapid initial respondents to the changing conditions with pigment markers showing an abrupt 4-fold increase in proportion but this increase was not sustained post-storm. 19’-HEX, a chemotaxonomic marker for prymnesiophytes, was the dominant accessory pigment pre- and post-storm with concentrations of 48-435 ng L-1, and represented 44% of total carotenoid concentrations. Accompanying scanning electron microscopy results support the pigment-based analysis but also provide detailed insight into the nano- and microplankton communities, which proved to be highly variable between pre-storm and post-storm sampling periods. Nanoplankton remained the dominant size class pre- and post-storm but the microplankton proportion peaked during the period of maximum nutrient and chlorophyll concentrations. Classic descriptions of autumn blooms resulting from storm driven eutrophication events promoting phytoplankton growth in surface waters should be tempered with greater understanding of the role of storm driven vertical reorganization of the water column and of resident phytoplankton communities. Crucially, in this case we observed no change in integrated chlorophyll, particulate organic carbon or biogenic silica concentrations despite also observing a ∼50% increase in surface chlorophyll concentrations which indicated that the surface enhancement in chlorophyll concentrations was most likely fed from below rather than resulting from in situ growth. Though not measured directly there was no evidence of enhanced export fluxes associated with this storm. These observations have implications for the growing practice of using chlorophyll fluorescence from remote platforms to determine ocean productivity late in the annual productivity period and in response to storm mixing

Role of RelA and SpoT in Burkholderia pseudomallei survival, biofilm formation and ceftazidime tolerance during nutritional stress

This is the author accepted manuscript.Burkholderia pseudomallei a saprophyte found in soil and stagnant water is the causative agent of human melioidosis, an often cause fatal disease. B. pseudomallei is intrinsically resistant to many antibiotics. The stringent response is a global bacterial adaptation process in response to nutritional limitation and is mediated by the alarmone (p)ppGpp, which is produced by two proteins, RelA and SpoT. In order to test whether the stringent response is involved in ceftazidime tolerance, biofilm formation, and bacterial survival in the soil microcosm, B. pseudomallei strain K96243 and its isogenic ΔrelA and ΔrelAΔspoT mutants were grown in rich and nutrient-limited media. In nutrient-limiting conditions, both the wild type and mutants were found to be up to 64-times more tolerant to ceftazidime than when grown in rich culture conditions. Moreover, the biofilm formation of all bacterial isolates tested were significantly higher under nutrient-limiting conditions than under nutrient-rich conditions. The ΔrelAΔspoT mutant produced less biofilm than its wild type or ΔrelA mutant under nutrient-limiting conditions. The survival of the ΔrelAΔspoT double mutant cultured in 1% moisture content soil was significantly decreased compared to the wild type and the ΔrelA mutant. Therefore, the RelA/SpoT protein family might represent a promising target for the development of novel antimicrobial agents to combat B. pseudomallei.This work was supported by the Thailand Research Fund through the Royal Golden Jubilee Ph.D. Program (Grant no. PHD/0351/2551 to CA and ST), the Higher Education Research Promotion and National Research University Project of Thailand, Office of the Higher Education Commission, through the Health Cluster (SHeP-GMS), and Khon Kaen University

Transcriptional profiles of Burkholderia pseudomallei reveal the direct and indirect roles of Sigma E under oxidative stress conditions

This is the final version of the article. Available from BioMed Central via the DOI in this record.BACKGROUND: Burkholderia pseudomallei, the causative agent of melioidosis, is a Gram-negative bacterium widely distributed in soil and water in endemic areas. This soil saprophyte can survive harsh environmental conditions, even in soils where herbicides (containing superoxide generators) are abundant. Sigma factor E (σE) is a key regulator of extra-cytoplasmic stress response in Gram-negative bacteria. In this study, we identified the B. pseudomallei σE regulon and characterized the indirect role that σE plays in the regulation of spermidine, contributing to the successful survival of B. pseudomallei in stressful environments. RESULTS: Changes in the global transcriptional profiles of B. pseudomallei wild type and σE mutant under physiological and oxidative stress (hydrogen peroxide) conditions were determined. We identified 307 up-regulated genes under oxidative stress condition. Comparison of the transcriptional profiles of B. pseudomallei wild type and σE mutant under control or oxidative stress conditions identified 85 oxidative-responsive genes regulated by σE, including genes involved in cell membrane repair, maintenance of protein folding and oxidative stress response and potential virulence factors such as a type VI secretion system (T6SS). Importantly, we identified that the speG gene, encoding spermidine-acetyltransferase, is a novel member of the B. pseudomallei σE regulon. The expression of speG was regulated by σE, implying that σE plays an indirect role in the regulation of physiological level of spermidine to protect the bacteria during oxidative stress. CONCLUSION: This study identified B. pseudomallei genes directly regulated by σE in response to oxidative stress and revealed the indirect role of σE in the regulation of the polyamine spermidine (via regulation of speG) for bacterial cell protection during oxidative stress. This study provides new insights into the regulatory mechanisms by which σE contributes to the survival of B. pseudomallei under stressful conditions.This work was supported by the National Science and Technology Development Agency and Siriraj Grant for Research and Development. S. Jitprasutwit was supported by the Royal Golden Jubilee Ph. D. Program (PHD0270/2551)

A content analysis of the consumer-facing online information about my health record: Implications for increasing knowledge and awareness to facilitate uptake and use

© The Author(s) 2017. Background: Low health literacy, low levels of positive belief and privacy and security concerns have been identified as a significant barrier to personal electronic health record uptake and use. An important tool for overcoming these barriers is the consumer-facing information which accompanies the system. My Health Record (MyHR) is the Australian national e-health record system, for which a large suite of online resources exists to facilitate consumer registration and use. This study uses a number of different measures of health resource quality to assess the MyHR online consumer-facing information and identify any gaps or areas for improvement. Objective: To analyse the quality and content of the online consumer-facing resources which support the uptake and use of MyHR. Method: Australian information resources aimed at healthcare consumers about the MyHR were included in this study. A comprehensive search using Internet search engines was conducted to locate all online consumer-facing resources about MyHR from both government and non-government sources. Readability (measured by Flesch–Kincaid grade level), year of publication/review, publishing organisation type, presentation style, linked websites, target audience, and themes were identified as important measures of health information quality, and these were recorded and reported on for each resource. Results: Eighty resources met the inclusion criteria. The mean Flesch–Kincaid grade level was 11.8. Most resources were created by Australian government sources (n = 55), and the most common target audience was the general public (n = 65). Registration (n = 51), privacy/security (n = 49), and benefits of use (n = 46) were the most common resource themes. Conclusion: The authors identified a number of gaps and areas for improvement in the provision of consumer-facing information about MyHR. Readability is too high for the general Australian population, and there are few translated resources, which means that the information provided does not cater to people with low literacy levels, communication disability, and/or difficulties in understanding written English. The target audiences for resources do not reflect priority groups that were identified during the MyHR development processes. There are also gaps in information provision about how consumers can use MyHR as a tool to meaningfully engage with health professionals and services to support their own person-centred care

Design and chemical evaluation of reduced machine-yield cigarettes

AbstractExperimental cigarettes (ECs) were made by combining technological applications that individually reduce the machine measured yields of specific toxicants or groups of toxicants in mainstream smoke (MS). Two tobacco blends, featuring a tobacco substitute sheet or a tobacco blend treatment, were combined with filters containing an amine functionalised resin (CR20L) and/or a polymer-derived, high activity carbon adsorbent to generate three ECs with the potential for generating lower smoke toxicant yields than conventional cigarettes. MS yields of smoke constituents were determined under 4 different smoking machine conditions. Health Canada Intense (HCI) machine smoking conditions gave the highest MS yields for nicotine-free dry particulate matter and for most smoke constituents measured. Toxicant yields from the ECs were compared with those from two commercial comparator cigarettes, three scientific control cigarettes measured contemporaneously and with published data on 120 commercial cigarettes. The ECs were found to generate some of the lowest machine yields of toxicants from cigarettes for which published HCI smoke chemistry data are available; these comparisons therefore confirm that ECs with reduced MS machine toxicant yields compared to commercial cigarettes can be produced. The results encourage further work examining human exposure to toxicants from these cigarettes, including human biomarker studies