The expression of potential molecular candidates for chloride ion channels in primary human granulocytes and granulocytic cell lines

INTRODUCTION AND HYPOTHESIS: The project aims to identify potential candidates for chloride (Cl-) ion channels in granulocytes, and granulocytic cell lines. It is hypothesised that Cl- ion channels, in particular hBest1, are implicated in the role of granulocytes in response to inflammation. METHODOLOGY: Two main methodologies were used; laboratory techniques and systematic review. Laboratory techniques included RT-PCR, flow cytometry and western blot analysis to characterise the expression of hANO1, hBest1 and hCLCA1 as potential chloride ion channels in granulocytes and granulocytic cell lines. Systematic review was performed to identify whether chloride ion channels are up-regulated in COPD and asthma. RESULTS: RT-PCR demonstrated hCLCA1 expression in granulocytes and eosinophils but not HL60. hBest1 and hBest3 was expressed in all 3 cell types. In granulocytes, flow cytometry demonstrated greater hCLCA1 protein expression intracellularly, compared to hBest1 protein, and greater hBest1 plasma membrane expression compared to hCLCA1 (P<0.05). There was a negative correlation between hBest1, and hCLCA1 but also a weak negative correlation between hBest1 and hANO1 (P<0.05). Granulocytes stimulated with IL-13 over 24 hours, had a greater protein expression both intracellularly and at the plasma membrane. There was increased migration of HL60s when transfected with hBest1, in response to fMLP (P<0.05). Systematic review did not support the project due to limitations. CONCLUSIONS: There is a complex relationship between hBest1, hCLCA1 and hANO1 which may contribute to the function of granulocytes. HBest1 protein expression peaked 24 hours after continuous stimulation with IL-13. This correlates with peak symptom expression in diseases such as COPD and asthma. It is suggested that hBest1 has a role in migration and activation of granulocytes, through regulation of cell shape and volume. It is concluded that hBest1 is a novel therapeutic target in the control of symptoms in chronic inflammatory lung diseases

Sociology, Science and Sustainability: Developing Relationships in Scotland

This paper considers the application of the sociological imagination during the analysis of data collected during an ethnographic study of an environmental regulator, the Scottish Environment Protection Agency (SEPA). SEPA is tasked with implementing the European Water Framework Directive (WFD) in Scotland, which will radically alter the regulation of water use. Applying a sociological imagination allowed the researcher to advocate for a more interdisciplinary and equitable understanding of sustainable water use when feeding back initial research results at the end of the data collection period. The researchers introduced socialised definitions of the environment, which linked social justice and ecological concerns. These insights provided a challenge to the traditional bio-physical science focus of the organisational participants, for whom sustainability is a relatively new addition to their duties. The paper concludes by discussing the importance of developing these interdisciplinary relationships in the future.Sociological Imagination, Water Framework Directive, Sustainability Science, Consumer-Citizens

The influence of cervical and thoracic lymphadenectomy on corneal allograft rejection in inbred rats

Aim To investigate the site of alloantigen presentation in the rat following orthotopic corneal transplantation. Methods Adult inbred Fischer 344 rats received penetrating corneal allografts from inbred Wistar Furth donors (n¼17), without lymphadenectomy. A second group (n¼8) underwent bilateral removal of superficial cervical and facial lymph nodes 7 days before transplantation. A third group (n¼9) underwent bilateral removal of superficial cervical, facial, internal jugular and posterior cervical nodes. Graft survival was assessed by corneal clarity and rejection was confirmed histologically. Results All allografts underwent rejection. The median time to rejection for unmodified allografts was day 15, compared with day 14.5 for minimally lymphadenectomised recipients and day 18 for more extensively lymphadenectomised recipients (p>0.05, all comparisons). The median day to rejection for the combined group of lymphadenectomised rats was day 17 (p>0.05 compared with unmodified grafts). The rejection process was similar in all recipients. Conclusions Removal of multiple lymph nodes in the neck and thorax did not significantly influence the incidence, tempo or nature of the corneal allograft response. Sensitisation and clonal expansion of corneal alloantigen-reactive cells cannot occur only in superficial cervical, facial, internal jugular and posterior cervical lymph nodes in the rat.We acknowledge financial support from the Australian National Health and Medical Research Council and the Ophthalmic Research Institute of Australia

Insights into the biodegradation of weathered hydrocarbons in contaminated soils by bioaugmentation and nutrient stimulation

Acknowledgements This work was supported by the LINK Bioremediation programme (BIOREM_35), the Environment Agency and the Biotechnology and Biological Sciences Research Council BBSRC (Grant BB/B512432/1). The authors also thank the UK Engineering and Physical Sciences Research Council (EPSRC) for financial support to carry out this work through a CASE award supported by the former FIRSTFARADAY (Environmental Sustainability KTN) partnership (Ref No. 5010978). The views expressed are authors’ alone and may not reflect the views or policies of their employing organisations.Peer reviewedPublisher PD

Data release notes : UK Geoenergy Observatories Glasgow Geothermal Energy Research Field Site (GGERFS) ground gas, 2018 and 2019 surveys

In 2014, the British Geological Survey (BGS) and the Natural Environment Research Council (NERC) were tasked with developing new centres for research into the sub-surface environment to aid the responsible development of new low-carbon energy technologies in the United Kingdom (UK) and internationally. Under the United Kingdom Geoenergy Observatories (UKGEOS) project, two sites were chosen, including the Glasgow Geothermal Energy Research Field Site (GGERFS) in the Cuningar Loop-Dalmarnock area in the east of Glasgow (Figure 1). The aims of the GGERFS facility include de-risking technical aspects of mine water geothermal to assess the feasibility of extracting/storing heat energy in an urbanised former coal mine setting (Monaghan 2019; Monaghan et al. 2017; Monaghan et al. 2018). The initial phase of the GGERFS project entails installing a network of boreholes into the superficial deposits and bedrock in the Cuningar Loop-Dalmarnock area of Glasgow to characterise the geological and hydrogeological setting and assess the potential for shallow geothermal energy. The borehole network is also designed for baseline monitoring to assess the environmental status before and during the lifetime of the project. A ground gas baseline is considered important at the GGERFS site to enable us to determine if there are significant ongoing ground gas contributions from sources such as (i) leakage from mine workings/features related to legacy mine workings (ii) gas generated from components of the made ground (building rubble, mine water, other waste) and (iii) natural soil processes. The made ground at Cuningar Loop is known to have been formed from a range of prior land uses (see Ramboll 2018 a, b) and is commonly around 10 m thick. Ground gas measurement is an important tool for monitoring geoenergy sites since sensitive measurements of, for example, CO2, CH4 and associated gases can be made directly within the biosphere in which we live. Monitoring of ground gas in the vadose zone has been undertaken as part of a broader GGERFS environmental monitoring effort that includes groundwater, soil and surface water chemistry, ground movement and seismicity. The intention of ground gas monitoring, indeed the environmental monitoring effort as a whole, is to characterise pre-existing i.e. pre-operational or baseline conditions, particularly with respect to former coal mining, building demolition, waste disposal/landfill, or other industrial activities, before significant development occurs in relation to GGERFS. As such, it should be noted that the August 2018 survey precedes any development of GGERFS and can be considered ‘baseline’ in the conventional sense, whereas the May and October 2019 surveys were conducted alongside site construction but ahead of site operation. Approaches to monitoring ground gas may include long term continuous monitoring using permanently deployed instruments, and discrete surveys involving mobile, wide area screening techniques (for example open path laser, cavity ring down laser) to augment high density grids of detailed point measurements. Point measurement data from ground gas surveys conducted at the Glasgow Geothermal Energy Research Field Site (GGERFS) in August 2018, and May and October 2019 are reported. Ground gas is defined here as: a. gas concentrations in the shallow (c.70-100 cm below ground level) unsaturated zone of the subsurface, and b. gas flux at the soil-atmosphere interfac

Expression of an anti-CD4 single-chain antibody fragment from the donor cornea can prolong corneal allograft survival in inbred rats

To investigate whether expression of an anti-CD4 antibody fragment (scFv) by a lentivector-transduced donor cornea can prolong rat corneal allograft survival. Methods Inbred Fischer 344 rats received penetrating corneal allografts from Wistar-Furth donors after a 3 h transduction of the donor cornea with a lentivector carrying anti-CD4scFv cDNA (Lv-CD4scFv), a lentivector carrying the reporter gene-enhanced yellow fluorescence protein (LV-eYFP), or an adenoviral vector carrying anti-CD4 scFv cDNA (Ad-CD4scFv). Unmodified controls were also performed. Graft survival was assessed by corneal clarity, and rejection was confirmed histologically. Results In organ-cultured corneas, expression of anti-CD4 scFv was detected at 2 days post-transduction with the adenoviral vector, compared with 5 days post-transduction with the lentivector, and was 10-fold higher than the former. More inflammation was observed in Ad-CD4scFv-modified allografts than in Lv-CD4scFv-modified grafts at 15 days postsurgery (p=0.01). The median time to rejection for unmodified, LV-eYFP and Ad-CD4scFv grafts was day 17, compared with day 22 for Lv-CD4scFv grafts (p≤0.018). Conclusion Donor corneas transduced with a lentiviral vector carrying anti-CD4scFv cDNA showed a modest but significant prolongation in graft survival compared with unmodified, Lv-eYFP and Ad-CD4scFv grafts. However, rejection still occurred in all Lv-CD4scFv grafts, indicating that sensitisation may have been delayed but was not prevented.Australian National Health & Medical Research Council and the Ophthalmic Research Institute of Australia

Changes in organic carbon to clay ratios in different soils and land uses in England and Wales over time

Realistic targets for soil organic carbon (SOC) concentrations are needed, accounting for differences between soils and land uses. We assess the use of SOC/clay ratio for this purpose by comparing changes over time in (a) the National Soil Inventory of England and Wales, first sampled in 1978–1983 and resampled in 1994–2003, and (b) two long-term experiments under ley-arable rotations on contrasting soils in the East of England. The results showed that normalising for clay concentration provides a more meaningful separation between land uses than changes in SOC alone. Almost half of arable soils in the NSI had degraded SOC/clay ratios ( 1/8, respectively. Given the wide range of soils and land uses across England and Wales in the datasets used to test these targets, they should apply across similar temperate regions globally, and at national to sub-regional scales.Biotechnology and Biological Sciences Research Council (BBSRC): BBS/E/C/000I0310 and BBS/E/C/000 J0300. Lawes Agricultural Trus

A social network analysis of social cohesion in a constructed pride: Implications for ex situ reintroduction of the African Lion (Panthera leo)

Animal conservation practices include the grouping of captive related and unrelated individuals to form a social structure which is characteristic of that species in the wild. In response to the rapid decline of wild African lion (Panthera leo) populations, an array of conservational strategies have been adopted. Ex situ reintroduction of the African lion requires the construction of socially cohesive pride structures prior to wild release. This pilot study adopted a social network theory approach to quantitatively assess a captive pride's social structure and the relationships between individuals within them. Group composition (who is present in a group) and social interaction data (social licking, greeting, play) was observed and recorded to assess social cohesion within a released semi-wild pride. UCINET and SOCPROG software was utilised to represent and analyse these social networks. Results indicate that the pride is socially cohesive, does not exhibit random associations, and the role of socially influential keystone individuals is important for maintaining social bondedness within a lion pride. These results are potentially informative for the structure of lion prides, in captivity and in the wild, and could have implications for captive and wild-founder reintroductions

Prevalence and architecture of de novo mutations in developmental disorders.

The genomes of individuals with severe, undiagnosed developmental disorders are enriched in damaging de novo mutations (DNMs) in developmentally important genes. Here we have sequenced the exomes of 4,293 families containing individuals with developmental disorders, and meta-analysed these data with data from another 3,287 individuals with similar disorders. We show that the most important factors influencing the diagnostic yield of DNMs are the sex of the affected individual, the relatedness of their parents, whether close relatives are affected and the parental ages. We identified 94 genes enriched in damaging DNMs, including 14 that previously lacked compelling evidence of involvement in developmental disorders. We have also characterized the phenotypic diversity among these disorders. We estimate that 42% of our cohort carry pathogenic DNMs in coding sequences; approximately half of these DNMs disrupt gene function and the remainder result in altered protein function. We estimate that developmental disorders caused by DNMs have an average prevalence of 1 in 213 to 1 in 448 births, depending on parental age. Given current global demographics, this equates to almost 400,000 children born per year

Design and validation of a multi-task, multi-context protocol for real-world gait simulation

Background: Measuring mobility in daily life entails dealing with confounding factors arising from multiple sources, including pathological characteristics, patient specific walking strategies, environment/context, and purpose of the task. The primary aim of this study is to propose and validate a protocol for simulating real-world gait accounting for all these factors within a single set of observations, while ensuring minimisation of participant burden and safety. Methods: The protocol included eight motor tasks at varying speed, incline/steps, surface, path shape, cognitive demand, and included postures that may abruptly alter the participants’ strategy of walking. It was deployed in a convenience sample of 108 participants recruited from six cohorts that included older healthy adults (HA) and participants with potentially altered mobility due to Parkinson’s disease (PD), multiple sclerosis (MS), proximal femoral fracture (PFF), chronic obstructive pulmonary disease (COPD) or congestive heart failure (CHF). A novelty introduced in the protocol was the tiered approach to increase difficulty both within the same task (e.g., by allowing use of aids or armrests) and across tasks. Results: The protocol proved to be safe and feasible (all participants could complete it and no adverse events were recorded) and the addition of the more complex tasks allowed a much greater spread in walking speeds to be achieved compared to standard straight walking trials. Furthermore, it allowed a representation of a variety of daily life relevant mobility aspects and can therefore be used for the validation of monitoring devices used in real life. Conclusions: The protocol allowed for measuring gait in a variety of pathological conditions suggests that it can also be used to detect changes in gait due to, for example, the onset or progression of a disease, or due to therapy. Trial registration: ISRCTN—12246987