Functional analysis of the leading malaria vaccine candidate AMA-1 reveals an essential role for the cytoplasmic domain in the invasion process

A key process in the lifecycle of the malaria parasite Plasmodium falciparum is the fast invasion of human erythrocytes. Entry into the host cell requires the apical membrane antigen 1 (AMA-1), a type I transmembrane protein located in the micronemes of the merozoite. Although AMA-1 is evolving into the leading blood-stage malaria vaccine candidate, its precise role in invasion is still unclear. We investigate AMA-1 function using live video microscopy in the absence and presence of an AMA-1 inhibitory peptide. This data reveals a crucial function of AMA-1 during the primary contact period upstream of the entry process at around the time of moving junction formation. We generate a Plasmodium falciparum cell line that expresses a functional GFP-tagged AMA-1. This allows the visualization of the dynamics of AMA-1 in live parasites. We functionally validate the ectopically expressed AMA-1 by establishing a complementation assay based on strain-specific inhibition. This method provides the basis for the functional analysis of essential genes that are refractory to any genetic manipulation. Using the complementation assay, we show that the cytoplasmic domain of AMA-1 is not required for correct trafficking and surface translocation but is essential for AMA-1 function. Although this function can be mimicked by the highly conserved cytoplasmic domains of P. vivax and P. berghei, the exchange with the heterologous domain of the microneme protein EBA-175 or the rhoptry protein Rh2b leads to a loss of function. We identify several residues in the cytoplasmic tail that are essential for AMA-1 function. We validate this data using additional transgenic parasite lines expressing AMA-1 mutants with TY1 epitopes. We show that the cytoplasmic domain of AMA-1 is phosphorylated. Mutational analysis suggests an important role for the phosphorylation in the invasion process, which might translate into novel therapeutic strategies.

Damming the flow: cultural barriers to perceived ‘procedural justice’ in Wonthaggi, Victoria

 Drier conditions in Australia have compelled governments to implement projects such as the desalination plant in the South Gippsland town of Wonthaggi. The desalination plant is still under construction, but South Gippsland is already host to wind turbines and marine protected areas, reflecting public pressure to develop renewable energy sources and conserve resources. However, all projects have been met with vocal opposition. Using the desalination project as a case study, this paper will address public concerns about a perceived lack of procedural justice in implementing such projects. Drawing on data from a pilot survey of 320 residents, we argue that procedural shortcomings of the project include inattention to past political disputes in the region and to the culturally entrenched sense of division between city and country. Attention to political and cultural histories is vital to the successful and ethical implementation of projects in regional areas.

Comparing towed and baited underwater video techniques for assessing temperate marine fishes

Accurate estimates of fish species occurrence are important to any species’ assessments and distribution model. With increasing emphasis on nondestructive sampling, underwater video techniques are commonly used without a thorough understanding of their advantages and disadvantages. This study compared data collected from baited remote underwater stereo-video systems (stereo BRUVS) and towed-video systems to determine; (1) the differences between these video techniques in terms of fish assemblages, functional groups (i.e. pelagic carnivore, epibenthic carnivore/omnivore or herbivore) and observability (i.e. conspicuous or cryptic), and (2) what impact do these two techniques have on the interpretation of spatially-explicit, predictive models. We found stereo BRUVS and towedvideo techniques recorded very different assemblages, functional groups and observability categories across structurally complex benthic biological habitats (i.e. macroalgae dominated habitats). However, as the habitat complexity became less (e.g. seagrass and areas with no visible macro-biota) both techniques appeared to provide similar fish assemblage information. We also found considerable differences in the predicted extents of habitat suitability between the two video techniques.

Integrating time-series hydroacoustics and video observations for detecting changes in benthic habitats

The ability to quantify change in marine benthic habitats must be considered a key goal of marine habitat mapping activities. Changes in distribution of distinct suites of benthic biological species may occur as a result of natural or human induced processes and these processes may operate at a range of temporal and spatial scales. It is important to understand natural small scale inter-annual patterns of change in order to separate these signals from potential patterns of longer term change. Work to describe these processes of change from an acoustic remote sensing stand point has thus far been limited due to the relatively recent availability of full coverage swath acoustic datasets and cost pressures associated with multiple surveys of the same area. This paper describes the use of landscape transition analysis as a means to differentiate seemingly random patterns of habitat change from systematic signals of habitat transition at a shallow (10–50 m depth) 18 km2 study area on the temperate Australian continental shelf between the years 2006 and 2007. Supervised classifications for each year were accomplished using independently collected high resolution (3 m cell-size) multibeam echosounder (MBES) and video-derived reference data. Of the 4 representative biotic classes considered, signals of directional systematic changes were observed to occur between a shallow kelp dominated class, a deep sessile invertebrate dominated class and a mixed class of kelp and sessile invertebrates. These signals of change are interpreted as inter-annual variation in the density and depth related extent of canopy forming kelp species at the site, a phenomenon reported in smaller scale temporal studies of the same species. The methods applied in this study provide a detailed analysis of the various components of the traditional change detection cross tabulation matrix allowing identification of the strongest signals of systematic habitat transitions across broad geographical regions. Identifying clear patterns of habitat change is an important first step in linking these patterns to the processes that drive them.

Testing the influence of survey method for building species distribution models of marine fishes

Accurate estimates of fish species occurrence are important to any species’ assessments and habitat suitability model. However, surveys of marine fishes are often biased by method. Surveys of marine fishes are often biased by method. Such bias could influence the interpretation of any habitat suitability model. With increasing emphasis on non-destructive sampling, underwater video techniques are commonly used without a thorough understanding of their advantages and disadvantages. This study compared data collected from baited remote underwater stereo-video systems and towed-video systems to provide occurrence data to develop habitat suitability models of nine temperate marine fishes. While numerous studies have compared modelling approaches in terms of model performance (i.e. via AUC or Kappa) the point of this paper was to highlight how very sensiblelooking, well-performing (based on AUC) models can provide different predictions of habitat suitability depending on which dataset is used.

Evaluating the travel, physical activity and carbon impacts of a 'natural experiment' in the provision of new walking and cycling infrastructure : methods for the core module of the iConnect study

Introduction Improving infrastructure to support walking and cycling is often regarded as fundamental to encouraging their widespread uptake. However, there is little evidence that specific provision of this kind has led to a significant increase in walking or cycling in practice, let alone wider impacts such as changes in overall physical activity or carbon emissions. Connect2 is a major new project that aims to promote walking and cycling in the UK by improving local pedestrian and cycle routes. It therefore provides a useful opportunity to contribute new evidence in this field by means of a natural experimental study.

Methods and analysis iConnect is an independent study that aims to integrate the perspectives of public health and transport research on the measurement and evaluation of the travel, physical activity and carbon impacts of the Connect2 programme. In this paper, the authors report the study design and methods for the iConnect core module. This comprised a cohort study of residents living within 5 km of three case study Connect2 projects in Cardiff, Kenilworth and Southampton, supported by a programme of qualitative interviews with key informants about the projects. Participants were asked to complete postal questionnaires, repeated before and after the opening of the new infrastructure, which collected data on demographic and socioeconomic characteristics, travel, car fuel purchasing and physical activity, and potential psychosocial and environmental correlates and mediators of those behaviours. In the absence of suitable no-intervention control groups, the study design drew on heterogeneity in exposure both within and between case study samples to provide for a counterfactual.

Ethics and dissemination The study was approved by the University of Southampton Research Ethics Committee. The findings will be disseminated through academic presentations, peer-reviewed publications and the study website (http://www.iconnect.ac.uk) and by means of a national seminar at the end of the study.

Interventions to promote cycling: a systematic review

Objectives To determine what interventions are effective in promoting cycling, the size of the effects of interventions, and evidence of any associated benefits on overall physical activity or anthropometric measures.

Design Systematic review.

Data sources Published and unpublished reports in any language identified by searching 13 electronic databases, websites, reference lists, and existing systematic reviews, and papers identified by experts in the field.

Review methods Controlled “before and after” experimental or observational studies of the effect of any type of intervention on cycling behaviour measured at either individual or population level.

Results Twenty five studies (of which two were randomised controlled trials) from seven countries were included. Six studies examined interventions aimed specifically at promoting cycling, of which four (an intensive individual intervention in obese women, high quality improvements to a cycle route network, and two multifaceted cycle promotion initiatives at town or city level) were found to be associated with increases in cycling. Those studies that evaluated interventions at population level reported net increases of up to 3.4 percentage points in the population prevalence of cycling or the proportion of trips made by bicycle. Sixteen studies assessing individualised marketing of “environmentally friendly” modes of transport to interested households reported modest but consistent net effects equating to an average of eight additional cycling trips per person per year in the local population. Other interventions that targeted travel behaviour in general were not associated with a clear increase in cycling. Only two studies assessed effects of interventions on physical activity; one reported a positive shift in the population distribution of overall physical activity during the intervention.

Conclusions Community-wide promotional activities and improving infrastructure for cycling have the potential to increase cycling by modest amounts, but further controlled evaluative studies incorporating more precise measures are required, particularly in areas without an established cycling culture. Studies of individualised marketing report consistent positive effects of interventions on cycling behaviour, but these findings should be confirmed using more robust study designs. Future research should also examine how best to promote cycling in children and adolescents and through workplaces. Whether interventions to promote cycling result in an increase in overall physical activity or changes in anthropometric measures is unclear.

Risk factors for Mycobacterium ulcerans infection, southeastern Australia

Buruli/Bairnsdale ulcer (BU) is a severe skin and soft tissue disease caused by Mycobacterium ulcerans. To better understand how BU is acquired, we conducted a case-control study during a sustained outbreak in temperate southeastern Australia. We recruited 49 adult patients with BU and 609 control participants from a newly recognized BU-endemic area in southeastern Australia. Participants were asked about their lifestyle and insect exposure. Odds ratios were calculated by using logistic regression and were adjusted for age and location of residence. Odds of having BU were at least halved for those who frequently used insect repellent, wore long trousers outdoors, and immediately washed minor skin wounds; odds were at least doubled for those who received mosquito bites on the lower legs or lower arms. This study provides new circumstantial evidence that implicates mosquitoes in the transmission of M. ulcerans in southeastern Australia.

A review of population-based prevalence studies of physical activity in adults in the Asia-Pacific region

Background: Physical activity (PA) surveillance is an important component of non-communicable disease risk factor monitoring, and occurs through national and international surveillance systems. This review identifies population PA estimates for adults in the Asia-Pacific region, and examines variation in trends and prevalence rates obtained using different PA measures.

Methods: Data were obtained from a MEDLINE search; World Health Organization’s Global Health Infobase; Government websites and reference lists of relevant papers. Inclusion criteria included: national studies or those reporting large scale population-level data; data published from 2000 to 2010 and trend data prior; sample sizes over n = 1000, or fewer subjects in small nations.

Results: In total, 56 population surveys from 29 Asia-Pacific countries were identified. Data on ‘sufficient physical activity’ amongst adults were available from 45 studies (80%), with estimates ranging from 7% to 93% (median 62%, inter-quartile range 40%-85%). For 14 countries, estimates of ‘sufficient activity’ were documented in multiple surveys using different methods, with the largest variation from 18% to 92% in Nepal. Median or mean METminutes/ day, reported in 20 studies, ranged from 6 to 1356. Serial trend data were available for 11 countries (22%), for periods spanning 2-10 years. Of these, five countries demonstrated increases in physical activity over time, four demonstrated decreases and three showed no changes.

Conclusions: Many countries in the Asia-Pacific region collect population-level PA data. This review highlights differences in estimates within and between countries. Some differences may be real, others due to variation in the PA questions asked and survey methods used. Use of standardized protocols and measures, and combined reporting of data are essential goals of improved international PA surveillance.

Toughening of a carbon-fibre composite using electrospun poly(Hydroxyether of Bisphenol A) nanofibrous membranes through inverse phase separation and inter-domain etherification

The interlaminar toughening of a carbon fibre reinforced composite by interleaving a thin layer (~20 microns) of poly(hydroxyether of bisphenol A) (phenoxy) nanofibres was explored in this work. Nanofibres, free of defect and averaging several hundred nanometres, were produced by electrospinning directly onto a pre-impregnated carbon fibre material (Toray G83C) at various concentrations between 0.5 wt % and 2 wt %. During curing at 150 °C, phenoxy diffuses through the epoxy resin to form a semi interpenetrating network with an inverse phase type of morphology where the epoxy became the co-continuous phase with a nodular morphology. This type of morphology improved the fracture toughness in mode I (opening failure) and mode II (in-plane shear failure) by up to 150% and 30%, respectively. Interlaminar shear stress test results showed that the interleaving did not negatively affect the effective in-plane strength of the composites. Furthermore, there was some evidence from DMTA and FT-IR analysis to suggest that inter-domain etherification between the residual epoxide groups with the pendant hydroxyl groups of the phenoxy occurred, also leading to an increase in glass transition temperature (~7.5 °C).

Structural characterisation and bioactivities of hybrid carrageenan-like sulphated galactan from red alga Furcellaria lumbricalis

The red alga, Furcellaria lumbricalis from the coast of the Prince Edward Island (PEI) in Atlantic Canada, was extracted with hot water and fractionated with 0.125. M KCl to obtain a carrageenan-like polysaccharide. The polysaccharide was further purified on ion-exchange and gel-permeation chromatography to yield a fraction (FB1) of uniform size and charge, with an average molecular weight of 428. kD. Oligosaccharides generated with acid hydrolysis of FB1 were sequenced using the electrospray ionisation collision induced dissociation tandem mass spectrometry (ES-CID-MS/MS) technique. On the basis of chemical and spectroscopic analysis, FB1 was characterised to be composed of 1,4-linked 3,6-anhydro-galactose (40%), 1,3-linked 4-sulphated-galactose (30%), 1,3-linked galactose (20%), 1,4-linked galactose (8%) and 1,4-linked 3-O-methyl-galactose (2%), which makes it be a novel sulphated galactan hybrid. The β-secretase (BACE) inhibition and immunomodulation activities of FB1-derived oligosaccharides were evaluated in vitro.

Developing a placebo-controlled trial in surgery : issues of design, acceptability and feasibility

Background: Surgical placebos are controversial. This in-depth study explored the design, acceptability, and feasibility issues relevant to designing a surgical placebo-controlled trial for the evaluation of the clinical and cost effectiveness of arthroscopic lavage for the management of people with osteoarthritis of the knee in the UK.
Methods: Two surgeon focus groups at a UK national meeting for orthopaedic surgeons and one regional surgeon focus group (41 surgeons); plenary discussion at a UK national meeting for orthopaedic anaesthetists (130 anaesthetists); three focus groups with anaesthetists (one national, two regional; 58 anaesthetists); two focus groups with members of the patient organisation Arthritis Care (7 participants); telephone interviews with people on consultant waiting lists from two UK regional centres (15 participants); interviews with Chairs of UK ethics committees (6 individuals); postal surveys of members of the British Association of Surgeons of the Knee (382 surgeons) and members of the British Society of Orthopaedic Anaesthetists (398 anaesthetists); two centre pilot (49 patients assessed).
Results: There was widespread acceptance that evaluation of arthroscopic lavage had to be conducted with a placebo control if scientific rigour was not to be compromised. The choice of placebo surgical procedure (three small incisions) proved easier than the method of anaesthesia (general anaesthesia). General anaesthesia, while an excellent mimic, was more intrusive and raised concerns among some stakeholders and caused extensive discussion with local decision-makers when seeking formal approval for the pilot. Patients were willing to participate in a pilot with a placebo arm; although some patients when allocated to surgery became apprehensive about the possibility of receiving placebo, and withdrew. Placebo surgery was undertaken successfully.
Conclusions: Our study illustrated the opposing and often strongly held opinions about surgical placebos, the ethical issues underpinning this controversy, and the challenges that exist even when ethics committee approval has been granted. It showed that a placebo-controlled trial could be conducted in principle, albeit with difficulty. It also highlighted that not only does a placebo-controlled trial in surgery have to be ethically and scientifically acceptable but that it also must be a feasible course of action. The place of placebo-controlled surgical trials more generally is likely to be limited and require specific circumstances to be met. Suggested criteria are presented.