• Enhancing the formulation of Shared Mental Models in football players

      Robbins, Mandy; Newton, Joseph L. (University of Chester, 2020-06-24)
      A significant amount of research has examined the development of Decision-Making (DM) in sport. However, only a limited amount of research has explored decisions in the context of which they are made. Using the Naturalistic Decision-Making (NDM) paradigm this thesis employs Cognitive Task Analysis (CTA) approaches and Performance Analysis (PA) within a professional football environment, through the lens of a performance analyst. Applying the DM Framework, outlined by Richards, Collins and Mascarenhas (2016), this thesis applied a mixed method approach using three CTA approaches; Critical Decision Method (CDM), Thinking Aloud Problem Solving (TAPS) and Concept Mapping (CM) to examine the development of Shared Mental Models (SMMs). Additionally, PA data was collected to measure on-field application of SMMs in the format of match performance. Method: The development of DM ability was examined using professional footballers (n=16) and professional coaches (n=2), over an eight-week period. PA provided video footage of critical attacking play situations for use in team meetings. Meetings were designed to empower players in the DM process and involved the integration of CTA approaches in the form of a DM booklet. The booklet consisted of questions and diagrams relating to six clips identified by the coaches. Players and coaches would reflect on the clips individually and collectively as a team in an off-field setting. Each clip was split into three Phases, and six clips were shown to all team members in weekly meetings. To ascertain the retention of SMMs developed over the eight weeks, qualitative narratives recorded by the players (DM booklets) were analysed. No CTA processes were applied on week seven, as this was classified as a retention week. The CTA booklet recorded individual team members understanding of the situation and facilitated group discussions after clips. PA analysed data in the form of match statistics to assess transference of SMMs to the field of play. Results: CTA analysed data qualitatively indicated that individual Situational Awareness (SA) improved. Players identified more key themes in weeks six and eight compared to week one, indicating a development in SMMs and increased compatibility of SMM outlined by the two expert coach’s SA. Additionally, the complexity of the players SMMs and team SMMs developed. The PA data illustrated that the team generated more shots on target and more shots on target per Phase 3 entry in weeks six and eight than week zero (pre-investigation). The improved on-field performance of key performance indicators, combined with the increased identification of key themes and growing compatibility of players SMM in line with the expert coaches, demonstrate a more developed SMM which resulted in enhanced DM by the team. In summary, the application of PA and CTA methods within an off-field environment provides a mechanism to develop SMMs in a professional football team which transfer to enhancing on-field team DM in competitive play. However, this work utilising off-field learning environments to enhance DM, is still in its early stages and more research is needed.
    • Spinal motor neurite outgrowth over glial scar inhibitors is enhanced by coculture with bone marrow stromal cells

      Wright, Karina; Johnson, William Eustace Basil; Uchida, Kenzo; Bara, Jennifer J.; Roberts, Sally; Masari, Wagih E.; Aston University; Keele University
      BACKGROUND CONTEXT: Transplantation of bone marrow cells into spinal cord lesions promotes functional recovery in animal models, and recent clinical trials suggest possible recovery also in humans. The mechanisms responsible for these improvements are still unclear. PURPOSE: To characterize spinal cord motor neurite interactions with human bone marrow stromal cells (MSCs) in an in vitro model of spinal cord injury (SCI). STUDY DESIGN/SETTING: Previously, we have reported that human MSCs promote the growth of extending sensory neurites from dorsal root ganglia (DRG), in the presence of some of the molecules present in the glial scar, which are attributed with inhibiting axonal regeneration after SCI. We have adapted and optimized this system replacing the DRG with a spinal cord culture to produce a central nervous system (CNS) model, which is more relevant to the SCI situation. METHODS: We have developed and characterized a novel spinal cord culture system. Human MSCs were cocultured with spinal motor neurites in substrate choice assays containing glial scar–associated inhibitors of nerve growth. In separate experiments, MSC-conditioned media were analyzed and added to spinal motor neurites in substrate choice assays. RESULTS: As has been reported previously with DRG, substrate-bound neurocan and Nogo-A repelled spinal neuronal adhesion and neurite outgrowth, but these inhibitory effects were abrogated in MSC/spinal cord cocultures. However, unlike DRG, spinal neuronal bodies and neurites showed no inhibition to substrates of myelin-associated glycoprotein. In addition, the MSC secretome contained numerous neurotrophic factors that stimulated spinal neurite outgrowth, but these were not sufficient stimuli to promote spinal neurite extension over inhibitory concentrations of neurocan or Nogo-A. CONCLUSIONS: These findings provide novel insight into how MSC transplantation may promote regeneration and functional recovery in animal models of SCI and in the clinic, especially in the chronic situation in which glial scars (and associated neural inhibitors) are well established. In addition, we have confirmed that this CNS model predominantly comprises motor neurons via immunocytochemical characterization. We hope that this model may be used in future research to test various other potential interventions for spinal injury or disease states
    • Early Transplantation of Mesenchymal Stem Cells After Spinal Cord Injury Relieves Pain Hypersensitivity Through Suppression of Pain-Related Signaling Cascades and Reduced Inflammatory Cell Recruitment

      Johnson, William Eustace Basil; Watanabe, Shuji; Uchida, Kenzo; Nakajima, Hideaki; Matsuo, Hideaki; Sugita, Daisuke; Yoshida, Ai; Honjoh, Kazuya; Baba, Hisatoshi; Aston University, University of Fukui
      Bone marrow-derived mesenchymal stem cells (BMSC) modulate inflammatory/immune responses and promote motor functional recovery after spinal cord injury (SCI). However, the effects of BMSC transplantation on central neuropathic pain and neuronal hyperexcitability after SCI remain elusive. This is of importance because BMSC-based therapies have been proposed for clinical treatment. We investigated the effects of BMSC transplantation on pain hypersensitivity in green fluorescent protein (GFP)-positive bone marrow-chimeric mice subjected to a contusion SCI, and the mechanisms of such effects. BMSC transplantation at day 3 post-SCI improved motor function and relieved SCI-induced hypersensitivities to mechanical and thermal stimulation. The pain improvements were mediated by suppression of protein kinase C-γ and phosphocyclic AMP response element binding protein expression in dorsal horn neurons. BMSC transplants significantly reduced levels of p-p38 mitogen-activated protein kinase and extracellular signal-regulated kinase (p-ERK1/2) in both hematogenous macrophages and resident microglia and significantly reduced the infiltration of CD11b and GFP double-positive hematogenous macrophages without decreasing the CD11b-positive and GFP-negative activated spinal-microglia population. BMSC transplants prevented hematogenous macrophages recruitment by restoration of the blood-spinal cord barrier (BSCB), which was associated with decreased levels of (a) inflammatory cytokines (tumor necrosis factor-α, interleukin-6); (b) mediators of early secondary vascular pathogenesis (matrix metallopeptidase 9); (c) macrophage recruiting factors (CCL2, CCL5, and CXCL10), but increased levels of a microglial stimulating factor (granulocyte-macrophage colony-stimulating factor). These findings support the use of BMSC transplants for SCI treatment. Furthermore, they suggest that BMSC reduce neuropathic pain through a variety of related mechanisms that include neuronal sparing and restoration of the disturbed BSCB, mediated through modulation of the activity of spinal-resident microglia and the activity and recruitment of hematogenous macrophages.
    • The developing landscape of diagnostic and prognostic biomarkers for spinal cord injury in cerebrospinal fluid and blood

      Hulme CH; Brown SJ; Fuller HR; Riddell J; Osman A; Chowdhury J; Kumar N; Johnson WE; Wright KT; Keele University, RJAH Orthopaedic Hospital, University of Glasgow, University of Chester (Nature Publishing Group, 2016-12-20)
      STUDY DESIGN: Review study. OBJECTIVES: The identification of prognostic biomarkers of spinal cord injury (SCI) will help to assign SCI patients to the correct treatment and rehabilitation regimes. Further, the detection of biomarkers that predict permanent neurological outcome would aid in appropriate recruitment of patients into clinical trials. The objective of this review is to evaluate the current state-of-play in this developing field. SETTING: Studies from multiple countries were included. METHODS: We have completed a comprehensive review of studies that have investigated prognostic biomarkers in either the blood or cerebrospinal fluid (CSF) of animals and humans following SCI. RESULTS: Targeted and unbiased approaches have identified several prognostic biomarkers in CSF and blood. These proteins associate with cellular damage following SCI and include components from neurons, oligodendrocytes and reactive astrocytes, that is, neurofilament proteins, glial fibrillary acidic protein, Tau and S100 calcium-binding protein β. Unbiased approaches have also identified microRNAs that are specific to SCI, as well as other cell damage-associated proteins. CONCLUSIONS: The discovery and validation of stable, specific, sensitive and reproducible biomarkers of SCI is a rapidly expanding field of research. So far, few studies have utilised unbiased approaches aimed at the discovery of biomarkers within the CSF or blood in this field; however, some targeted approaches have been successfully used. Several studies using various animal models and some with small human patient cohorts have begun to pinpoint biomarkers in the CSF and blood with putative prognostic value. An increased sample size will be required to validate these biomarkers in the heterogeneous clinical setting.
    • The influence of pH and fluid dynamics on the antibacterial efficacy of 45S5 Bioglass Short title: Antibacterial efficacy of 45S5 Bioglass

      Begum, Saima; Johnson, William Eustace Basil; Worthington, Tony; Martin, Richard; Aston University (IOP Publishing, 2016-02-02)
      In recent years, there has been considerable interest in the potential antibacterial properties that bioactive glasses may possess. However, there have been several conflicting reports on the antibacterial efficacy of 45S5 Bioglass®. Various mechanisms regarding its mode of action have been proposed, such as changes in the environmental pH, increased osmotic pressure, and 'needle-like' sharp glass debris which could potentially damage prokaryotic cell walls and thus inactivate bacteria. In this current study, a systematic investigation was undertaken on the antibacterial efficacy of 45S5 Bioglass® on Escherichia coli NCTC 10538 and Staphylococcus aureus ATCO 6538 under a range of clinically relevant scenarios including varying Bioglass® concentration, direct and indirect contact between Bioglass® and microorganisms, static and shaking incubation conditions, elevated and neutralised pH environments. The results demonstrated that, under elevated pH conditions, Bioglass® particles have no antibacterial effect on S. aureus while a concentration dependent antibacterial effect against E. coli was observed. However, the antibacterial activity ceased when the pH of the media was neutralised. The results of this current study, therefore, suggest that the mechanism of antibacterial activity of Bioglass® is associated with changes in the environmental pH; an environment that is less likely to occur in vivo due to buffering of the system.
    • Bone Marrow-Derived Mesenchymal Stem Cells Become Antiangiogenic When Chondrogenically or Osteogenically Differentiated: Implications for Bone and Cartilage Tissue Engineering

      Bara, Jennifer; Johnson, William Eustace Basil; Roberts, Sally; McCarthy, Helen E.; Humphrey, Emma; AO Institute, Davos, Switzerland; Aston University; Keele University
      Osteochondral tissue repair requires formation of vascularized bone and avascular cartilage. Mesenchymal stem cells stimulate angiogenesis both in vitro and in vivo but it is not known if these proangiogenic properties change as a result of chondrogenic or osteogenic differentiation. We investigated the angiogenic/antiangiogenic properties of equine bone marrow-derived mesenchymal stem cells (eBMSCs) before and after differentiation in vitro. Conditioned media from chondrogenic and osteogenic cell pellets and undifferentiated cells was applied to endothelial tube formation assays using Matrigel. Additionally, the cell secretome was analysed using LC-MS/MS mass spectrometry and screened for angiogenesis and neurogenesis-related factors using protein arrays. Endothelial tube-like formation was supported by conditioned media from undifferentiated eBMSCs. Conversely, chondrogenic and osteogenic conditioned media was antiangiogenic as shown by significantly decreased length of endothelial tube-like structures and degree of branching compared to controls. Undifferentiated cells produced higher levels of angiogenesis-related proteins compared to chondrogenic and osteogenic pellets. In summary, eBMSCs produce an array of angiogenesis-related proteins and support angiogenesis in vitro via a paracrine mechanism. However, when these cells are differentiated chondrogenically or osteogenically, they produce a soluble factor(s) that inhibits angiogenesis. With respect to osteochondral tissue engineering, this may be beneficial for avascular articular cartilage formation but unfavourable for bone formation where a vascularized tissue is desired.
    • CD271-selected mesenchymal stem cells from adipose tissue enhance cartilage repair and are less angiogenic than plastic adherent mesenchymal stem cells

      Kohli, Nupur; Johnson, William Eustace Basil; Uchida, Kenzo; Aston University, University of Chester, University of Fukui (Nature, 2019-02-28)
      CD271 is a marker of bone marrow MSCs with enhanced differentiation capacity for bone or cartilage repair. However, the nature of CD271+ MSCs from adipose tissue (AT) is less well understood. Here, we investigated the differentiation, wound healing and angiogenic capacity of plastic adherent MSCs (PA MSCs) versus CD271+ MSCs from AT. There was no difference in the extent to which PA MSCs and CD271+ MSCs formed osteoblasts, adipocytes or chondrocytes in vitro. In contrast, CD271+ MSCs transplanted into athymic rats significantly enhanced osteochondral wound healing with reduced vascularisation in the repair tissue compared to PA MSCs and control animals; there was little histological evidence of mature articular cartilage formation in all animals. Conditioned medium from CD271+ MSC cultures was less angiogenic than PA MSC conditioned medium, and had little effect on endothelial cell migration or endothelial tubule formation in vitro. The low angiogenic activity of CD271+ MSCs and improved early stage tissue repair of osteochondral lesions when transplanted, along with a comparable differentiation capacity along mesenchymal lineages when induced, suggests that these selected cells are a better candidate than PA MSCs for the repair of cartilaginous tissue.
    • An In Vitro Comparison of the Incorporation, Growth, and Chondrogenic Potential of Human Bone Marrow versus Adipose Tissue Mesenchymal Stem Cells in Clinically Relevant Cell Scaffolds Used for Cartilage Repair

      Kohli, Nupur; Johnson, William Eustace Basil; Wright, Karina T.; Sammons, Rachel L.; Jeys, Lee; Snow, Martyn
      Aim: To compare the incorporation, growth, and chondrogenic potential of bone marrow (BM) and adipose tissue (AT) mesenchymal stem cells (MSCs) in scaffolds used for cartilage repair. Methods: Human BM and AT MSCs were isolated, culture expanded, and characterised using standard protocols, then seeded into 2 different scaffolds, Chondro-Gide or Alpha Chondro Shield. Cell adhesion, incorporation, and viable cell growth were assessed microscopically and following calcein AM/ethidium homodimer (Live/Dead) staining. Cell-seeded scaffolds were treated with chondrogenic inducers for 28 days. Extracellular matrix deposition and soluble glycosaminoglycan (GAG) release into the culture medium was measured at day 28 by histology/immunohistochemistry and dimethylmethylene blue assay, respectively. Results: A greater number of viable MSCs from either source adhered and incorporated into Chondro-Gide than into Alpha Chondro Shield. In both cell scaffolds, this incorporation represented less than 2% of the cells that were seeded. There was a marked proliferation of BM MSCs, but not AT MSCs, in Chondro-Gide. MSCs from both sources underwent chondrogenic differentiation following induction. However, cartilaginous extracellular matrix deposition was most marked in Chondro-Gide seeded with BM MSCs. Soluble GAG secretion increased in chondrogenic versus control conditions. There was no marked difference in GAG secretion by MSCs from either cell source. Conclusion: Chondro-Gide and Alpha Chondro Shield were permissive to the incorporation and chondrogenic differentiation of human BM and AT MSCs. Chondro-Gide seeded with BM MSCs demonstrated the greatest increase in MSC number and deposition of a cartilaginous tissue.
    • Comparison of Mesenchymal Stromal Cells Isolated From Murine Adipose Tissue and Bone Marrow in the Treatment of Spinal Cord Injury

      Takahashi, Ai; Johnson, William Eustace Basil; Uchida, Kenzo; Matsumine, Akihiko; University of Chester, University of Fukui (SAGE, 2018-05-10)
      The use of mesenchymal stromal cell (MSC) transplantation to repair the injured spinal cord has shown consistent benefits in preclinical models. However, the low survival rate of grafted MSC is one of the most important problems. In the injured spinal cord, transplanted cells are exposed to hypoxic conditions and exposed to nutritional deficiency caused by poor vascular supply. Also, the transplanted MSCs face cytotoxic stressors that cause cell death. The aim of this study was to compare adipose-derived MSCs (AD-MSCs) and bone marrow-derived MSCs (BM-MSCs) isolated from individual C57BL6/J mice in relation to: (i) cellular characteristics, (ii) tolerance to hypoxia, oxidative stress and serum-free conditions, and (iii) cellular survival rates after transplantation. AD-MSCs and BM-MSCs exhibited a similar cell surface marker profile, but expressed different levels of growth factors and cytokines. To research their relative stress tolerance, both types of stromal cells were incubated at 20.5% O2 or 1.0% O2 for 7 days. Results showed that AD-MSCs were more proliferative with greater culture viability under these hypoxic conditions than BM-MSCs. The MSCs were also incubated under H2O2-induced oxidative stress and in serum-free culture medium to induce stress. AD-MSCs were better able to tolerate these stress conditions than BMMSCs; similarly when transplanted into the spinal cord injury region in vivo, AD-MSCs demonstrated a higher survival rate post transplantation Furthermore, this increased AD-MSC survival post transplantation was associated with preservation of axons and enhanced vascularization, as delineated by increases in anti-gamma isotype of protein kinase C and CD31 immunoreactivity, compared with the BM-MSC transplanted group. Hence, our results indicate that AD-MSCs are an attractive alternative to BM-MSCs for the treatment of severe spinal cord injury. However, it should be noted that the motor function was equally improved following moderate spinal cord injury in both groups, but with no significant improvement seen unfortunately following severe spinal cord injury in either group
    • Canine mesenchymal stem cells are neurotrophic and angiogenic: An in vitro assessment of their paracrine activity.

      Johnson, William Eustace Basil; Al Delfi, Ibtesam; Aston University, University of Chester, Veterinary Tissue Bank Ltd (Elsevier, 2016-09-19)
      Mesenchymal stem cells (MSCs) have been used in cell replacement therapies for connective tissue damage, but also can stimulate wound healing through paracrine activity. In order to further understand the potential use of MSCs to treat dogs with neurological disorders, this study examined the paracrine action of adipose-derived canine MSCs on neuronal and endothelial cell models. The culture-expanded MSCs exhibited a MSC phenotype according to plastic adherence, cell morphology, CD profiling and differentiation potential along mesenchymal lineages. Treating the SH-SY5Y neuronal cell line with serum-free MSC culture-conditioned medium (MSC CM) significantly increased SH-SY5Y cell proliferation (P <0.01), neurite outgrowth (P = 0.0055) and immunopositivity for the neuronal marker βIII-tubulin (P = 0.0002). Treatment of the EA.hy926 endothelial cell line with MSC CM significantly increased the rate of wound closure in endothelial cell scratch wound assays (P = 0.0409), which was associated with significantly increased endothelial cell proliferation (P <0.05) and migration (P = 0.0001). Furthermore, canine MSC CM induced endothelial tubule formation in EA.hy926 cells in a soluble basement membrane matrix. Hence, this study has demonstrated that adipose-derived canine MSC CM stimulated neuronal and endothelial cells probably through the paracrine activity of MSC-secreted factors. This supports the use of canine MSC transplants or their secreted products in the clinical treatment of dogs with neurological disorders and provides some insight into possible mechanisms of action.
    • The Development and Growth of Tissues Derived From Cranial Neural Crest and Primitive Mesoderm Is Dependent on the Ligation Status of Retinoic Acid Receptor γ: Evidence That Retinoic Acid Receptor γ Functions to Maintain stem/progenitor Cells in the Absence of Retinoic Acid

      Johnson, William Eustace Basil; Wai, Htoo Aung; Aston University (Mary Ann Liebert, Inc, 2015-02-15)
      Retinoic acid (RA) signaling is important to normal development. However, the function of the different RA receptors (RARs)--RARα, RARβ, and RARγ--is as yet unclear. We have used wild-type and transgenic zebrafish to examine the role of RARγ. Treatment of zebrafish embryos with an RARγ-specific agonist reduced somite formation and axial length, which was associated with a loss of hoxb13a expression and less-clear alterations in hoxc11a or myoD expression. Treatment with the RARγ agonist also disrupted formation of tissues arising from cranial neural crest, including cranial bones and anterior neural ganglia. There was a loss of Sox 9-immunopositive neural crest stem/progenitor cells in the same anterior regions. Pectoral fin outgrowth was blocked by RARγ agonist treatment. However, there was no loss of Tbx-5-immunopositive lateral plate mesodermal stem/progenitor cells and the block was reversed by agonist washout or by cotreatment with an RARγ antagonist. Regeneration of the caudal fin was also blocked by RARγ agonist treatment, which was associated with a loss of canonical Wnt signaling. This regenerative response was restored by agonist washout or cotreatment with the RARγ antagonist. These findings suggest that RARγ plays an essential role in maintaining stem/progenitor cells during embryonic development and tissue regeneration when the receptor is in its nonligated state.
    • Retention of cardiac rehabilitation services during the COVID-19 pandemic A joint position statement from the British Association for Cardiovascular Prevention and Rehabilitation (BACPR) British Cardiovascular Society (BCS) British Heart Foundation (BHF)

      Dawkes, Susan; Hughes, Sally; Ray, Simon; Nichols, Simon; Hinton, Sally; Roberts, Ceri; Butler, Thomas; Delal, Hayes; Docherty, Patrick; University of York/National Audit of Cardiac Rehabilitation
      The British Association for Cardiovascular Prevention and Rehabilitation (BACPR), the British Cardiovascular Society (BCS) and the British Heart Foundation (BHF) have issued a joint position statement ‘Retention of cardiac rehabilitation services during the COVID-19 pandemic’.
    • Open Carboniferous Limestone pavement grike microclimates in Great Britain and Ireland: understanding the present to inform the future

      Burek, Cynthia; Hosie, Lottie; Geary, Matt; York, Peter, J. (University of ChesterUniversity of Chester, 2020-04)
      Limestone pavements are a distinctive and irreplaceable geodiversity feature, in which are found crevices known as grikes. These grikes provide a distinct microclimate conferring a more stable temperature, higher relative humidity, lower light intensity and lower air speed than can be found in the regional climate. This stability of microclimate has resulted in an equally distinctive community of flora and fauna, adapted to a forest floor but found in an often otherwise barren landscape. This thesis documents the long-term study of the properties of the limestone pavement grike in order to identify the extent to which the microclimate may sustain its distinctive biodiversity, to provide recommendations for future research which may lead to more effective management. Over a five-year study, recordings of temperature, relative humidity, light intensity and samples of invertebrate biodiversity were collected from five limestone pavements situated in the Yorkshire Dales and Cumbria in Great Britain, and The Burren in the Republic of Ireland. An extensive description of the grike microclimate was undertaken using the data collected to understand the extent of the microclimate stability of the grike and the conditions for variation in the grike microclimate. Further insights into the grike microclimate were gained through simulation techniques more commonly used in engineering, to explore the effects of air flowing over a grike, the light from the sun entering the grike and regression analysis to simulate the temperature within the grike in the present and projected for the future. This study has indicated that although the whole of the grike confers a degree of microclimatic stability, it is made up of a less stable upper zone and a more stable lower zone. The instability of the upper zone is hypothesised to result from the extent to which the majority of light and external air can enter the grike, whereas the stability of the lower zone may be governed by the thermal stability of the limestone surrounding it. Based on this zonation and the projections for the grike temperature, it is hypothesised that climate change will have the most substantial effects in the upper grike zone where species obligated to this area could be most heavily impacted. This study recommends a range of areas in which research may be employed so that the limestone pavement habitat may be successfully managed in Great Britain and the Republic of Ireland.
    • The importance of long-term genetic monitoring of reintroduced populations: inbreeding in the natterjack toad (Epidalea calamita)

      Muir, Anna P; orcid: 0000-0002-6896-6915; Phillips, Susanna; Geary, Matt; email: m.geary@chester.ac.uk; Allmark, Matthew; Bennett, Sarah; Norman, Kim; Ball, Rachel J; Peters, Catherine; University of Chester; Cheshire Wildlife Trust; Eni UK Ltd
      Genetic monitoring is an important, but frequently lacking, component of management actions to support long-term persistence in reintroduced populations. Populations that remain small, due to demographic processes and genetic diversity, are more likely to experience a second extinction event. The natterjack toad (Epidelea calamita) is legally protected in Britain and was the subject of a reintroduction programme in the 1990s. However, subsequent genetic assessment has been mostly lacking. The aim of this study was to assess the genetic diversity of two reintroduced populations of natterjack toads in order to inform conservation management. Adults were sampled and nine microsatellites amplified to assess neutral genetic variation within each site and for comparison with the source population. Inbreeding was observed at the reintroduction sites, as evidenced by high FIS values (0.43 and 0.72), low observed compared to expected heterozygosities, and significant deviation from Hardy-Weinberg equilibrium. Observed heterozygosity is currently lower in the reintroduction sites than it was in the source population at the time of the reintroductions (Red Rocks: 0.15±0.20; Talacre: 0.12±0.20; Ainsdale (source): 0.29). Evidence for a bottleneck was not found, although this is likely a result of sampling overlapping generations. No within-site population structuring was observed. Such low genetic diversity has not previously been recorded in any natterjack population. Genetic rescue, combined with pool creation, is the most viable option for safeguarding the species at these sites into the future. Our work highlights the importance of ongoing genetic monitoring, in collaboration with conservation organisations, to support conservation management.
    • Factors influencing dietary behaviours in urban food environments in Africa: a systematic mapping review.

      Osei-Kwasi, Hibbah; orcid: 0000-0001-5084-6213; Mohindra, Aarti; Booth, Andrew; Laar, Amos; Wanjohi, Milka; Graham, Fiona; Pradeilles, Rebecca; Cohen, Emmanuel; orcid: 0000-0001-5643-1473; Holdsworth, Michelle; orcid: 0000-0001-6028-885X (2020-05-26)
      To identify factors influencing dietary behaviours in urban food environments in Africa and identify areas for future research. We systematically reviewed published/grey literature (protocol CRD4201706893). Findings were compiled into a map using a socio-ecological model on four environmental levels: individual, social, physical and macro. Urban food environments in Africa. Studies involving adolescents and adults (11-70 years, male/female). Thirty-nine studies were included (six adolescent, fifteen adolescent/adult combined and eighteen adult). Quantitative methods were most common (twenty-eight quantitative, nine qualitative and two mixed methods). Studies were from fifteen African countries. Seventy-seven factors influencing dietary behaviours were identified, with two-thirds at the individual level (45/77). Factors in the social (11/77), physical (12/77) and macro (9/77) environments were investigated less. Individual-level factors that specifically emerged for adolescents included self-esteem, body satisfaction, dieting, spoken language, school attendance, gender, body composition, pubertal development, BMI and fat mass. Studies involving adolescents investigated social environment-level factors more, for example, sharing food with friends. The physical food environment was more commonly explored in adults, for example, convenience/availability of food. Macro-level factors associated with dietary behaviours were food/drink advertising, religion and food prices. Factors associated with dietary behaviour were broadly similar for men and women. The dominance of studies exploring individual-level factors suggests a need for research to explore how social, physical and macro-level environments drive dietary behaviours of adolescents and adults in urban Africa. More studies are needed for adolescents and men, and studies widening the geographical scope to encompass all African countries.
    • Investigating the Prevalence of Anaemia in Rural Gambia, in Relation to Levels of Zinc Protoporphyrin, Haemoglobin and Haptoglobin (Phenotype and Genotype)

      Bah, Ebrima; Michelangeli, Frank (Oxford University Press (OUP), 2020-05-29)
      Abstract Objectives To find out the overlapping and correlating relationships between serum haptoglobin level, haptoglobin genotype and phenotype, blood haemoglobin level and zinc protoporphyrin (measured in washed RBCs) in association to prevalence of anaemia. It will focus on comparing all the mention components in contrast to each other. The study will also look for the frequency distribution of the major HP alleles. Methods 1278 participants were randomly selected. Blood samples collected by trained nurses. Data generation was done at the Medical research council (keneba field station) research site. Data Analysis was conducted at the university of Chester with the assistance of the computer department team. Results P = 0.000 indicating anaemia prevalence with HP 1 allele. P &amp;gt; 0.05 when ID, IDA and AI relates with HP genotype. Positive correlation between ZnPP and HP serum level, but negative between ZnPP and Hb. P = 0.000 between ZnPP and IDA. P = 0.024 between HP genotype and Hb level. P = 0.013 between HP genotype and HP serum. P = 0.100 between HP genotype and ZnPP. P = 0.000 between ZnPP and IDA. P = 0.024 between HP genotype and Hb. ZnPP shared a positive correlation with HP serum level, and a negative correlation with Hb level. The correlation significant = 0.01 level (2-tailed) P = 0.01. The correlation between HP genotype and HP serum level was significant with P = 0.013, but the correlation between HP genotype and ZnPP was not significant with P = 0.100. Conclusions HP genotype had association with anaemia prevalence and more occurrence was observed in carriers of the type ‘1’ allele. It had no association with ID, IDA and AI. HP genotype had association with HP serum level and Hb level but had no association with ZnPP level. ZnPP level was observed to have had association with HP serum level, Hb level and IDA; but had no association with ID and AI in the region. Funding Sources All the resources used in this study were from MRC Keneba (International Nutrition Group) which is supported by funds from the UK Medical Research Council (MRC) and the UK Department for International Development (DFID) under the MRC/DFID Concordat agreement (Hennig et al., 2015).
    • Patterns in island endemic forest-dependent bird research: the Caribbean as a case-study

      Devenish-Nelson, Eleanor S.; orcid: 0000-0002-9029-4772; email: ellie.devenish@ed.ac.uk; Weidemann, Douglas; Townsend, Jason; Nelson, Howard P. (Springer Netherlands, 2019-05-04)
      Abstract: Unequal patterns in research effort can result in inaccurate assessments of species extinction risk or ineffective management. A group of notable conservation concern are tropical island endemic birds, many of which are also forest-dependent, which increases their vulnerability to extinction. Yet, island bird species have received limited research attention compared to their continental congeners, despite this taxon being globally regarded as well-studied. We used the insular Caribbean, a globally important endemism hotspot with high rates of deforestation, to explore research bias of island and regional endemic forest-dependent birds. A review of the published literature (n = 992) found no significant increase in the number of studies over the search period. Research effort was significantly higher among species with threatened status, long generation time, wide habitat breadth and low to intermediate elevational distributions. Among family groups, the Psittacidae received the highest research effort, while the Cuculidae were the most underrepresented family (30-fold higher and six-fold less than expected, respectively). We found geographic biases in effort, with Jamaica having six-fold less and Puerto Rico eight times more research than expected for their level of endemism. These patterns likely reflect individual interests and limited capacity and funding, typical of Small Island Developing States. With over 50% of species in this review having declining population trends, we recommend prioritizing research that emphasises conservation- and management-relevant data across underrepresented families and islands, by fostering greater collaboration between researchers, practitioners and the existing local amateur ornithological community.
    • The Functionality of “Ogi” – a fermented cereal gruel made in Nigeria, in the Management of Gastrointestinal Disease

      Pickles, Neil; Weili, Li; Olayanju, Adetokunbo T (University of ChesterUniversity of Chester, 2019-04)
      Fermentation is an ancient food processing technique which has over the years had significant impact on food availability, production, economy and health. Fermented corn, ‘ogi’, is a popular meal in South and West African countries and is also traditionally administered to people, especially children, during the incidence of diarrhoea as it is believed to enhance gut health. Various authors have supported the advantageous health functions of fermented corn but have focussed on the possible health benefits that may be derived from the action of the fermentation organisms by investigating their probiotic potentials. Furthermore, in vivo studies have investigated effective functionality of fermented foods and where studies have suggested positive effect of the fermented food intervention, the component of the complex food matrix that may be responsible is often unknown. Though hypothesised, there has been little information on the possible effects of the end products of the fermentation on gut health. Therefore, this thesis aimed to explore the functionality potentials for ‘ogi’, to enhance gut health by investigating the mechanisms through which it may be able to confer the health benefits when consumed. An independent meta-analysis was undertaken to explore and review the analysis of the effectiveness of fermented foods to significantly reduce the duration of diarrhoea. The first experimental study was a comparison between spontaneously fermented vs controlled method of ‘ogi’ production with the aim to explore the possibility of achieving similar end products. The probiotic potential of fermentation organisms and antimicrobial effect of fermentation organisms against E. coli were also investigated. Colonic short chain fatty acids (SCFAs) are known to enhance immune function in the gut. The second experimental study explored the production of SCFAs in fermented corn with the theory that fermented corn may be a dietary source of SCFA and may produce a comparable effect in the gut as the colonic SCFA. It is established that one of the functions of endogenously synthesised nitric oxide (NO) in the body is to enhance immune functions. Therefore, the potential for ‘ogi’ to enhance immune response in the gut by being a dietary source of Nitric Oxide (NO) was explored in the third experimental study. The outcomes of this thesis suggest that it may not be possible to mimic the exact spontaneous fermentation process in a controlled setting. Weissella confuse, Pediococcus pentosaceus and Saccharomyces cerevisiae were the lactic acid bacteria (LAB) and yeast isolated from spontaneously fermented corn using 16S rRNA sequencing. The isolated organisms along with Lactobacillus plantarum were used as starter culture for controlled fermentation. There was a significant difference between the end products of both fermentations. All organisms except W. confuse showed probiotic potential. Acetate was produced during both fermentations but butyrate was produced during spontaneous fermentation only. There was however higher concentration of NO in controlled fermented corn than spontaneous fermentation with nitrate values of 30.54 µM and 23.21 µM and nitrite values of 17.50 µM and 10.11 µM respectively. These suggest that fermented corn may have probiotic potential and may be able to further enhance immune health in the gut by being a dietary source of SCFAs and NO.
    • An investigation of canine mesenchymal stem cells and their secretome in the context of spinal cord injury

      Johnson, Eustace; Wood, Chelsea R (University of Chester, 2020-05-26)
      Spinal cord injury (SCI) is a condition that has devastating effects on both humans and animals alike. Damage inflicted causes loss of neural tissue and secondary inflammatory mechanisms produce an inhibitory environment that results in partial or complete loss of motor and sensory functions. Additionally, SCI can cause multisystem issues such as organ failures, infections, muscle atrophy and decrease in mental health. Coupled with emotional and financial burdens, these effects can reduce quality of life. Mesenchymal Stem Cells (MSC) are known to have immunomodulatory, angiogenic and paracrine activity, all of which are beneficial to wound healing following SCI. Pre-clinical studies have shown encouraging results of MSC therapy for SCI, however replication of results has been difficult to achieve in the clinic. Dogs also suffer from SCI and show the same heterogenous nature and pathophysiology of SCI as humans. This provides a good potential clinical model for MSC therapies for SCI, as well as providing benefit in the veterinary clinic. Therefore, the overall aim of this study was to assess if canine MSC (cMSC) and cMSC secretome (conditioned medium; CM) could potentially be used for treatment of SCI in veterinary clinics, simultaneously providing model data that could be translated into the human clinic. It was first required to confirm efficacy of cMSC when used to treat other conditions in dogs, such as arthritis, along with safety of autologous transplantation. Characterisation of both cMSC phenotype and paracrine (angiogenic and neurogenic) activity was confirmed using ISCT criteria and the established cell lines EA.hy926 and SH-SY5Y. Further examination showed that exposure to certain elements of the injured spinal cord, such as CSPG which are found within the inhibitory glial scar, exerted some effects on cMSC and cMSC angiogenic and neurogenic paracrine activity. To finish, the study aimed to assess the effect of cMSC CM on an ex vivo model of the spinal cord, a multicellular environment and it was found that cMSC CM increased astrocyte reactivity but reduced neuronal maturation and growth, suggesting that cMSC paracrine activity depends in part on the spinal cord microenvironment. Overall, this study has shown that cMSC, in particular cMSC CM, could be used as complete or partial treatment for SCI in dogs.
    • Mangrove or mudflat: Prioritising fish habitat for conservation in a turbid tropical estuary

      Marley, Guy; Deacon, Amy; Philip, Dawn; Lawrence, Andrew; University of the West Indies, University of the West Indies, University of the West Indies, University of Chester
      Mangrove habitats are typically the focus of conservation efforts in tropical estuaries because their structural complexity is thought to support greater biodiversity and nursery function than unvegetated habitats. However, evidence for this paradigm has been equivocal in turbid tropical estuaries where unvegetated mudflats are also highly productive. The present study compared the community composition, biodiversity, nursery-role and commercial fish biomass in two mangrove habitats and one mudflat habitat in the Gulf of Paria, Trinidad. A total of 12 705 fishes, comprising 63 species from 26 families, were sampled in mangrove creeks, seaward mangrove fringe and the subtidal margin of an intertidal mudflat from June 2014 to June 2015. The composition of the creek and mudflat communities were distinct, while the community of the mangrove fringe more closely resembled the mudflat than the mangrove creeks. Mean species richness (MSR), total species richness (TSR) extrapolated from species accumulation curves, and juvenile species richness (JSR) were significantly greater in the mudflat (MSR = 11.4 ± 1.0; TSR = 75 ± 14; JSR = 9.1 ± 0.8) than mangrove creeks (MSR = 9.0 ± 0.5; TSR = 49 ± 3; JSR = 6.1 ± 0.4) and the seaward mangrove fringe (MSR = 6.4 ± 0.7; TSR = 58 ± 14; JSR = 5.2 ± 0.4). Meanwhile, Shannon Weiner diversity, juvenile fish abundance and commercial fish biomass were comparable between habitats. These findings caution against the generalisation that mangroves are the most important habitat for fishes in turbid tropical estuaries. There is now a growing body of evidence that mudflats warrant consideration as important repositories of biodiversity and nursery function for juvenile fishes.