The In Vitro Antioxidant and Anti-Inflammatory Activities of Selected Australian Seagrasses.

Recent studies have shown that seagrasses could possess potential applications in the treatment of inflammatory disorders. Five seagrass species (Zostera muelleri, Halodule uninervis, Cymodocea rotundata, Syringodium isoetifolium, and Thalassia hemprichii) from the Great Barrier Reef (QLD, Australia) were thus collected, and their preliminary antioxidant and anti-inflammatory activities were evaluated. From the acetone extracts of five seagrass species subjected to 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging antioxidant assay, the extract of Z. muelleri had the highest activity (half minimal concentration of inhibition (IC50) = 138 µg/mL), with the aerial parts (IC50 = 119 µg/mL) possessing significantly higher antioxidant activity than the roots (IC50 ≥ 500 µg/mL). A human peripheral blood mononuclear cells (PBMCs) assay with bacterial lipopolysaccharide (LPS) activation and LEGENDplex cytokine analysis showed that the aerial extract of Z. muelleri significantly reduced the levels of inflammatory cytokines tumour necrosis factor alpha (TNF-α), interleukin (IL)-1ß, and IL-6 by 29%, 74%, and 90%, respectively, relative to the LPS treatment group. The aerial extract was thus fractionated with methanol (MeOH) and hexane fraction, and purification of the MeOH fraction by HPLC led to the isolation of 4-hydroxybenzoic acid (1), luteolin (2), and apigenin (3) as its major constituents. These compounds have been previously shown to reduce levels of TNF-α, IL-1ß, and IL-6 and represent some of the major bioactive components of Z. muelleri aerial parts. This investigation represents the first study of the antioxidant and anti-inflammatory properties of Z. muelleri and the first isolation of small molecules from this species. These results highlight the potential for using seagrasses in treating inflammation and the need for further investigation.

Inner Nuclear Membrane Asi Ubiquitin Ligase Catalytic Subunits Asi1p and Asi3p, but not Asi2p, confer resistance to aminoglycoside hygromycin B in Saccharomyces cerevisiae.

The heterotrimeric Asi ubiquitin ligase (encoded by ASI1, ASI2, and ASI3) mediates protein degradation in the inner nuclear membrane in Saccharomyces cerevisiae. Asi1p and Asi3p possess catalytic domains, while Asi2p functions as an adaptor for a subset of Asi substrates. We hypothesized the Asi complex is an important mediator of protein quality control, and we predicted that Asi would be required for optimal growth in conditions associated with elevated abundance of aberrant proteins. Loss of Asi1p or Asi3p, but not Asi2p, sensitized yeast to hygromycin B, which promotes translational infidelity by distorting the ribosome A site. Surprisingly, loss of quality control ubiquitin ligase Hul5p did not sensitize yeast to hygromycin B. Our results are consistent with a prominent role for an Asi subcomplex that includes Asi1p and Asi3p (but not Asi2p) in protein quality control.

Green turtle (Chelonia mydas) grazing plot formation creates structural changes in a multi-species Great Barrier Reef seagrass meadow.

The Great Barrier Reef (GBR) contains extensive seagrass meadows with abundant and diverse herbivore populations. Typically, meadows in the region are multi-species and dominated by fast growing opportunistic seagrass species. However, we know little about how herbivores modify these types of seagrass meadows by grazing. We conducted the first megaherbivore exclusion study in the GBR at Green Island (Queensland) to understand how green turtle grazing structures these multi-species tropical seagrass meadows. After excluding green turtles for three months, we found that grazing only impacted seagrasses at one site, where green turtles created a grazing plot by actively feeding on both above and below ground seagrass structures, a rare observation for the species. Within this grazing plot at the end of the experiment, the un-caged control treatments open to grazing had a 60% reduction in both above and below ground biomass, and shoot height was reduced by 75%, but there was no impact of grazing on the seagrass species mix. Our study shows that grazing plot formation by green turtles occurs in GBR fast growing seagrass communities and reduces both above and below ground seagrass biomass, this behaviour may be targeting elevated leaf nutrients, or nutritional content of rhizomes. This study is the first documented case of grazing plot formation by green turtles in the GBR and suggests that grazing pressure has a major influence on seagrass meadow structure.

Transient Receptor Potential Ankyrin-1 and Vanilloid-3 Differentially Regulate Endoplasmic Reticulum Stress and Cytotoxicity in Human Lung Epithelial Cells After Pneumotoxic Wood Smoke Particle Exposure.

This study investigated the roles of transient receptor potential (TRP) ankyrin-1 (TRPA1) and TRP vanilloid-3 (TRPV3) in regulating endoplasmic reticulum stress (ERS) and cytotoxicity in human bronchial epithelial cells (HBECs) treated with pneumotoxic wood smoke particulate matter (WSPM) and chemical agonists of each channel. Functions of TRPA1 and TRPV3 in pulmonary epithelial cells remain largely undefined. This study shows that TRPA1 activity localizes to the plasma membrane and endoplasmic reticulum (ER) of cells, whereas TRPV3 resides primarily in the ER. Additionally, treatment of cells using moderately cytotoxic concentrations of pine WSPM, carvacrol, and other TRPA1 agonists caused ERS as a function of both TRPA1 and TRPV3 activities. Specifically, ERS and cytotoxicity were attenuated by TRPA1 inhibition, whereas inhibiting TRPV3 exacerbated ERS and cytotoxicity. Interestingly, after treatment with pine WSPM, TRPA1 transcription was suppressed, whereas TRPV3 was increased. TRPV3 overexpression in HBECs conferred resistance to ERS and an attenuation of ERS-associated cell cycle arrest caused by WSPM and multiple prototypical ERS-inducing agents. Alternatively, short hairpin RNA-mediated knockdown of TRPV3, like the TRPV3 antagonist, exacerbated ERS. This study reveals previously undocumented roles for TRPA1 in promoting pathologic ERS and cytotoxicity elicited by pneumotoxic WSPM and TRPA1 agonists, and a unique role for TRPV3 in fettering pathologic facets of the integrated ERS response. SIGNIFICANCE STATEMENT: These findings provide new insights into how wood smoke particulate matter and other transient receptor potential ankyrin-1 (TRPA1) and transient receptor potential vanilloid-3 (TRPV3) agonists can affect human bronchial epithelial cells and highlight novel physiological and pathophysiological roles for TRPA1 and TRPV3 in these cells.

Temporal correspondence of selenium and mercury, among brine shrimp and water in Great Salt Lake, Utah, USA.

The specific source of high burdens of selenium (Se) and mercury (Hg) in several bird species at Great Salt Lake (GSL) remain unknown. Frequent co-located water and brine shrimp samples were collected during 2016 through 2017 to identify potential correlations of element concentrations among brines and brine shrimp, a keystone species in the GSL. Like many aquatic systems, GSL is characterized by elevated methylmercury (MeHg) in deep waters. However, in contrast to thermally-stratified aquatic systems, biota in the salinity-stratified GSL do not reside in its deep waters, obscuring the presumed relationship between elevated MeHg in biota and in the deep brine. Brine shrimp and water column (shallow and deep, filtered and unfiltered) samples were collected from six sites spanning the South Arm of GSL approximately every other month. Mercury concentrations in brine shrimp (on average 89% of which is MeHg) were correlated only with total mercury in surface filtered water, and displayed little spatial variability, but consistent seasonal trends across the two sampled years. In contrast to Hg, temporal correspondence was observed between Se concentrations in brine shrimp and those in all water samples regardless of filtering and depth, with maxima and minima at higher-than-seasonal frequency. The data suggest a spatially diffuse source of bioavailable mercury to the shallow brine that responds to seasonal influences, for which the underlying deep brine, surficial sediments, and overlying atmosphere were evaluated in terms of potential temporal correspondence to shallow brine and brine shrimp Hg concentrations, as well as potential to mix across the extent of the shallow brine. Bioaccumulation factors were at the low end of those reported for marine systems, and decreased at higher trace element concentrations in water.

Wood Smoke Particles Stimulate MUC5AC Overproduction by Human Bronchial Epithelial Cells Through TRPA1 and EGFR Signaling.

Mucus hypersecretion is a pathological feature of acute inflammatory and chronic obstructive pulmonary diseases. Exposure to air pollutants can be a cause of pathological mucus overproduction, but mechanisms by which different forms of air pollutants elicit this response are not fully understood. In this study, particulate matter (PM) generated from burning pine wood and other types of biomass was used to determine mechanisms by which these forms of PM stimulate mucin gene expression and secretion by primary human bronchial epithelial cells (HBECs). Biomass PM < 2.5 µm generated from pine wood and several other fuels stimulated the expression and secretion of the gel-forming glycoprotein MUC5AC by HBECs. Muc5ac gene induction was also observed in mouse airways following subacute oropharyngeal delivery of pine wood smoke PM. In HBECs, MUC5AC was also induced by the transient receptor potential ankyrin-1 (TRPA1) agonists' coniferaldehyde, a component of pine smoke PM, and allyl isothiocyanate, and was attenuated by a TRPA1 antagonist. Additionally, inhibition of epidermal growth factor receptor (EGFR/ErbB1) and the EGFR signaling partners p38 MAPK and GSK3ß also prevented MUC5AC overexpression. Collectively, our results suggest that activation of TRPA1 and EGFR, paired with alterations to p38 MAPK and GSK3ß activity, plays a major role in MUC5AC overproduction by bronchial epithelial cells exposed to biomass smoke PM. These results reveal specific processes for how biomass smoke PM may impact the human respiratory system and highlight potential avenues for therapeutic manipulation of lung diseases that are affected by air pollutants.

Wood and Biomass Smoke: Addressing Human Health Risks and Exposures.

Air pollutants from burning wood and biomass pose serious human health risks. Recent discoveries link the chemistry of smoke emissions with biochemical sensors and biological effect mediators. Strategic thinking of complex underlying factors is needed to protect people from harm.

The Role of Herbivory in Structuring Tropical Seagrass Ecosystem Service Delivery.

Seagrass meadows support key ecosystem services, via provision of food directly for herbivores, and indirectly to their predators. The importance of herbivores in seagrass meadows has been well-documented, but the links between food webs and ecosystem services in seagrass meadows have not previously been made explicit. Herbivores interact with ecosystem services - including carbon sequestration, cultural values, and coastal protection. Interactions can be positive or negative and depend on a range of factors including the herbivore identity and the grazing type and intensity. There can be unintended consequences from management actions based on a poor understanding of trade-offs that occur with complex seagrass-herbivore interactions. Tropical seagrass meadows support a diversity of grazers spanning the meso-, macro-, and megaherbivore scales. We present a conceptual model to describe how multiple ecosystem services are influenced by herbivore pressure in tropical seagrass meadows. Our model suggests that a balanced ecosystem, incorporating both seagrass and herbivore diversity, is likely to sustain the broadest range of ecosystem services. Our framework suggests the pathway to achieve desired ecosystem services outcomes requires knowledge on four key areas: (1) how size classes of herbivores interact to structure seagrass; (2) desired community and management values; (3) seagrass responses to top-down and bottom-up controls; (4) the pathway from intermediate to final ecosystem services and human benefits. We suggest research should be directed to these areas. Herbivory is a major structuring influence in tropical seagrass systems and needs to be considered for effective management of these critical habitats and their services.

Developing nurse medication safety training in a health partnership in Mozambique using behavioural science.

BACKGROUND: Globally, safe and effective medication administration relies on nurses being able to apply strong drug calculation skills in their real-life practice, in the face of stressors and distractions. These may be especially prevalent for nurses in low-income countries such as Mozambique and Continuing Professional Development post-registration may be important. This study aimed to 1) explore the initial impact of an international health partnership's work to develop a drug calculation workshop for nurses in Beira, Mozambique and 2) reflect upon the role of health psychologists in helping educators apply behavioural science to the training content and evaluation. METHODS: In phase one, partners developed a training package, which was delivered to 87 Portuguese-speaking nurses. The partnership's health psychologists coded the training's behaviour change content and recommended enhancements to content and delivery. In phase two, the refined training, including an educational game, was delivered to 36 nurses in Mozambique and recoded by the health psychologists. Measures of participant confidence and intentions to make changes to healthcare practice were collected, as well as qualitative data through post-training questions and 12 short follow-up participant interviews. RESULTS: In phase one six BCTs were used during the didactic presentation. Most techniques targeted participants' capability to calculate drug doses accurately; recommendations aimed to increase participants' motivation and perceived opportunity, two other drivers of practice change. Phase two training included an extra seven BCTs, such as action planning and further skills practice. Participants reported high confidence before and after the training (p = 0.25); intentions to use calculators to check drug calculations significantly increased (p = 0.031). Qualitative data suggested the training was acceptable, enjoyable and led to practice changes, through improved capability, opportunity and motivation. Opportunity barriers to medication safety were highlighted. CONCLUSIONS: Reporting and measuring medication errors and related outcomes is a complex challenge affecting global efforts to improve medication safety. Through strong partnership working, a multi-disciplinary team of health professionals including health psychologists developed, refined and begin to evaluate a locally-led drug calculation CPD workshop for nurses in a low-resource setting. Applying behavioural science helped to collect feasible evaluation data and hopefully improved impact and sustainability.

Teaching Quality Improvement in Graduate Medical Education: An Experiential and Team-Based Approach to the Acquisition of Quality Improvement Competencies.

PROBLEM: An emerging priority in medical education is the need to facilitate learners' acquisition of quality improvement (QI) competencies. Accreditation bodies in both Canada and the United States have included QI and patient safety in their core competencies. APPROACH: In 2010, the Department of Family Medicine at Queen's University designed a graduate medical education curriculum to engage residents in a clinical QI program that would meet accreditation requirements. Monthly didactic sessions were combined with an experiential, team-based QI project that aligned with existing clinic priorities. The curriculum spans the first year of residency and is divided into three stages: (1) Engaging, (2) Understanding, and (3) Improving and translating. In Stage 1, teams of residents select a clinical QI topic, engage stakeholders, and collect baseline data related to their topic. In Stage 2, they focus on understanding their problem, interpreting their results, and applying QI tools. In Stage 3, they develop change ideas, translate their knowledge, and prepare to hand over their project. OUTCOMES: This QI curriculum aided residents in effectively acquiring QI competencies and allowed them to experience real-world challenges, such as securing project buy-in, negotiating with peers, and developing solutions to problems. Unlike in many QI programs, residents learned how to improve quality rather than about QI; thus, they formed the necessary foundation to carry out QI work in the future. NEXT STEPS: The curriculum will be evaluated using a knowledge assessment and satisfaction tool and postproject resident interviews. Facilitators will focus more on improving faculty develop ment in QI.

Empower U: effectiveness of an adolescent outreach and prevention program with sixth-grade boys and girls: a pilot study.

Sixth graders are at a prime age to modify behaviors and beliefs regarding exercise, nutrition, body image, and smoking. Empower U was created to change knowledge, beliefs, and behaviors regarding these topics. This pilot study utilized pre/post assessments of 58 sixth graders from a private middle school in the midsouth. Results showed a significant increase in self-esteem as well as in exercise and nutrition knowledge and beliefs at posttest and a significant increase in body image as well as in self-reported exercise and nutrition behaviors at the 1-month follow-up. Empower U provides nurses with an effective educational program that may be useful in positively impacting health behaviors.

Statistical selection of maintenance genes for normalization of gene expressions.

Maintenance genes can be used for normalization in the comparison of gene expressions. Even though the absolute expression levels of maintenance genes may vary considerably among different tissues or cells, a set of maintenance genes may provide suitable normalization if their expression levels are relatively constant in the specific tissues or cells of interest. A statistical procedure is proposed to select maintenance genes for normalization of gene expression data from tissues or cells of interest. This procedure is based on simultaneous confidence intervals for practical equivalence of relative gene expressions in these tissues or cells. As an illustration, the procedure is applied to the maintenance gene expression data from Vandesompele et al. (2002).

CGHScan: finding variable regions using high-density microarray comparative genomic hybridization data.

BACKGROUND: Comparative genomic hybridization can rapidly identify chromosomal regions that vary between organisms and tissues. This technique has been applied to detecting differences between normal and cancerous tissues in eukaryotes as well as genomic variability in microbial strains and species. The density of oligonucleotide probes available on current microarray platforms is particularly well-suited for comparisons of organisms with smaller genomes like bacteria and yeast where an entire genome can be assayed on a single microarray with high resolution. Available methods for analyzing these experiments typically confine analyses to data from pre-defined annotated genome features, such as entire genes. Many of these methods are ill suited for datasets with the number of measurements typical of high-density microarrays. RESULTS: We present an algorithm for analyzing microarray hybridization data to aid identification of regions that vary between an unsequenced genome and a sequenced reference genome. The program, CGHScan, uses an iterative random walk approach integrating multi-layered significance testing to detect these regions from comparative genomic hybridization data. The algorithm tolerates a high level of noise in measurements of individual probe intensities and is relatively insensitive to the choice of method for normalizing probe intensity values and identifying probes that differ between samples. When applied to comparative genomic hybridization data from a published experiment, CGHScan identified eight of nine known deletions in a Brucella ovis strain as compared to Brucella melitensis. The same result was obtained using two different normalization methods and two different scores to classify data for individual probes as representing conserved or variable genomic regions. The undetected region is a small (58 base pair) deletion that is below the resolution of CGHScan given the array design employed in the study. CONCLUSION: CGHScan is an effective tool for analyzing comparative genomic hybridization data from high-density microarrays. The algorithm is capable of accurately identifying known variable regions and is tolerant of high noise and varying methods of data preprocessing. Statistical analysis is used to define each variable region providing a robust and reliable method for rapid identification of genomic differences independent of annotated gene boundaries.