• Androgens in a female primate: relationships with reproductive status, age, dominance rank, fetal sex and secondary sexual color

      Setchell, Joanna M.; Smith, Tessa E.; Knapp, Leslie A.; Durham University; University of Chester; University of Cambridge (Elsevier, 2015-05-01)
      A comprehensive understanding of the role of androgens in reproduction, behavior andmorphology requires the examination of female, aswell as male, hormone profiles. However, we know far less about the biological significance of androgens in females than in males. We investigated the relationships between fecal androgen (immunoreactive testosterone) levels and reproductive status, age, dominance rank, fetal sex and a secondary sexual trait (facial color) in semi-free-ranging femalemandrills (Mandrillus sphinx), using samples collected from19 reproductively mature females over 13 months. Fecal androgens varied with reproductive status, being highest during gestation. Fecal androgens began to increase at 3 months of gestation, and peaked at 5 months. This pattern is more similar to that found in a platyrrhine than in other cercopithecine species, suggesting that such patterns are not necessarily phylogenetically constrained. Fecal androgens did not vary systematically with rank, in contrast to the relationship we have reported for male mandrills, and in line with sex differences in how rank is acquired and maintained. Offspring sex was unrelated to fecal androgens, either prior to conception or during gestation, contrasting with studies of other primate species. Mean facial color was positively related to mean fecal androgens across females, reflecting the same relationship inmalemandrills. However, the relationship between color and androgens was negative within females. Future studies of the relationship between female androgens and social behavior, reproduction and secondary sexual traits will help to elucidate the factors underlying the similarities and differences found between the sexes and among studies.
    • Animal Social Network Theory Can Help Wildlife Conservation

      Snijders, Lysanne; Blumstein, Daniel; Franks, Daniel Wayne; Stanley, Christina R.; University of Chester; Leibniz-Institute of Freshwater Ecology and Inland Fisheries, Berlin; Wageningen University & Research; University of California; University of York (Elsevier, 2017-06-22)
      Many animals preferentially associate with certain other individuals. This social structuring can influence how populations respond to changes to their environment, thus making network analysis a promising technique for understanding, predicting and potentially manipulating population dynamics. Various network statistics can correlate with individual fitness components and key population-level processes, yet the logical role and formal application of animal social network theory for conservation and management have not been well articulated. We outline how understanding of direct and indirect relationships between animals can be profitably applied by wildlife managers and conservationists. By doing so, we aim to stimulate the development and implementation of practical tools for wildlife conservation and management and to inspire novel behavioral research in this field.
    • ANNING, Mary (1799-1847)

      Burek, Cynthia V.; University College Chester (Thoemmes Continuum, 2004-06-01)
      This dictionary entry discusses the life and work of British fossil hunter Mary Anning (1799-1847).
    • Anthropometric and physical performance characteristics of top-elite, elite and non-elite youth female team handball players

      Moss, Samantha L.; McWhannell, Nicola; Michalsik, Lars B.; Twist, Craig (2015-02-16)
      In order to maximise the potential for success, developing nations need to produce superior systems to identify and develop talent, which requires comprehensive and up-to-date values on elite players. This study examined the anthropometric and physical characteristics of youth female team handball players (16.07 ± 1.30 y) in non-elite (n= 47), elite (n= 37) and top-elite players (n= 29). Anthropometric profiling included sum of eight skinfolds, body mass, stature, girths, breadths and somatotype. Performance tests included 20 m sprint, counter movement jump, throwing velocity, repeated shuttle sprint and jump ability test, and Yo-Yo Intermittent Recovery Test Level 1. Youth top-elite players had greater body mass, lean mass, stature, limb girths and breadths than elite and non-elite players, while only stature and flexed arm were higher in elite compared to non-elite players (all P < 0.05). Sum of skinfolds and waist-to-hip ratio were similar between groups (P > 0.05). Top-elite performed better in most performance tests compared to both elite and non-elite players (P < 0.05), although maximal and repeated10 m sprints were similar between standard (P > 0.05). Elite outperformed non-elite players in throwing velocity only. Findings reveal that non-elite players compare unfavourably to top-elite international European players in many anthropometric and performance characteristics, and differ in few characteristics compared to elite European club team players. This study is useful for emerging team handball nations in improving talent identification processes.
    • Anti-epileptic drugs and bone loss: phenytoin reduces pro-collagen I and alters the electrophoretic mobility of osteonectin in cultured bone cells.

      Wilson, Emma L.; Garton, Mark; Fuller, Heidi R.; RJAH Orthopaedic Hospital; RJAH Orthopaedic NHS Foundation Trust; Keele University (Elsevier, 2016-05-31)
      Phenytoin is an antiepileptic drug used in the management of partial and tonic-clonic seizures. In previous studies we have shown that valproate, another antiepileptic drug, reduced the amount of two key bone proteins, pro-collagen I and osteonectin (SPARC, BM-40), in both skin fibroblasts and cultured osteoblast-like cells. Here we show that phenytoin also reduces pro-collagen I production in osteoblast-like cells, but does not appear to cause a decrease in osteonectin message or protein production. Instead, a 24h exposure to a clinically relevant concentration of phenytoin resulted in a dose-dependent change in electrophoretic mobility of osteonectin, which was suggestive of a change in post-translational modification status. The perturbation of these important bone proteins could be one of the mechanisms to explain the bone loss that has been reported following long-term treatment with phenytoin.
    • Antibacterial activity of Manuka honey and its components: An overview

      Johnston, Matthew; McBride, Michael; Dahiya, Divakar; Owusu-Apenten, Richard K.; Nigam, Poonam S.; University of Chester, University of Ulster (AIMS Press, 2018-11-27)
      The importance of honey for medicinal purposes is well documented in some of the world’s oldest literature. Honey is well known and studied for its antimicrobial properties. The medicinal properties in honey originate from the floral source used by bees. Manuka honey is a dark monofloral honey rich in phenolic content, and currently it is gaining much attention for its antimicrobial activity. Researchers have found that honey is effective against a wide range of pathogens. The antibacterial potency of Manuka honey was found to be related to the Unique Manuka Factor (UMF) rating, which is correlated with the methylglyoxal and total phenols content. It is reported that different types of Manuka honey have differing effects and Gram-negative bacteria are more resistant than Gram-positive bacteria. Bacterial resistance to honey as antimicrobial agent has yet to be identified, possibly due to the presence of a complex mixture of methylglyoxal and other components. Honey is also reported to alter a bacterium’s shape and size through septal ring alteration, which affects cell morphology and growth. Research has shown that Manuka honey of different UMF values has medicinal properties of interest and it can be beneficial when used as a combination treatment with other antimicrobial agents
    • Antioxidant and genoprotective activity of selected cucurbitaceae seed extracts and LC–ESIMS/MS identification of phenolic components

      Yasir, Muhammad; Sultana, Bushra; Nigam, Poonam S.; Owusu-Apenten, Richard K.; University of Agriculture, Faisalabad, Pakistan; University of California; Ulster University (Elsevier, 2015-11-30)
      Cucurbitaceae are one of most widely used plant species for human food but lesser known members have not been examined for bioactive components. The purpose of this study was to evaluate the antioxidant and genoprotective activities from three cucurbitaceae seeds extracts and to identify phenolic components by LC–ESIMS/MS analysis. From the results, the yield of seeds extract was 20–41% (w/w) and samples had 16–40% total phenols as gallic acid equivalents (GAE). Compared with methanol solvent, using acidified methanol led to increased extraction yield by 1.4 to 10-fold, higher phenolic content (149.5 ± 1.2 to 396.4 ± 1.9 mg GAE/g), higher DPPH radical quenching and enhanced enoprotective activity using the pBR322 plasmid assay. LC–ESI-MS/MS analysis led to identification of 14–17 components, based on authentic standards and comparison with literature reports, as mainly phenolic acids and esters, flavonol glycosides. This may be the first mass spectrometric profiling of polyphenol components from cucurbitaceae seeds.
    • Antioxidant, Anticancer and Antibacterial Activity of Withania somnifera Aqueous Root Extract

      Barnes, D. A.; Barlow, R.; Nigam, Poonam S.; Owusu-Apenten, Richard K.; University of Chester, University of Ulster (Sciencedomain international, 2015-11-10)
      Aims: To evaluate total antioxidant capacity, anticancer activity and antibacterial effects Withania somnifera aqueous-root extracts. Study Design: In vitro study. Place of Study: School of Biomedical Sciences, Ulster University, UK. Methodology: Total antioxidant capacity (TAC) of whole powder and freeze dried W. somnifera aqueous-root extracts was determined using FRAP, DPPH, Folin and ABTS assays. Anticancer activity was accessed using MDA-MB-231 breast cells and Sulforhodamine B staining for cell viability. Antibacterial activity was by disk diffusion assay with penicillin, amoxicillin and streptomycin as positive controls. Results: The TAC for W. somnifera extract was 86, 47, 195,or 443 gallic acid equivalents per 100g dry basis (mgGAE/ 100 g) using FRAP, DPPH, Folin or ABTS assays, respectively. Corresponding TAC values for freeze dried W. somnifera aqueous-root extract were, 418, 553, 1898 or, 1770 (mgGAE/100 g). W. somnifera aqueous-root extract inhibited MDA-MB-231 cell proliferation in a dose-dependent manner with IC50 = 0.19 mg/ml (21 µM GAE). Nil antibacterial effects were detected for freeze dried W. somnifera extract (0-1 mg/ml) across six species of bacteria tested. Conclusion: Withania somnifera root water extract showed significant antioxidant and anticancer activity for MDA-MB-231 breast cancer cells but no antibacterial activity under the conditions of this study.
    • Antioxidant, Anticancer and Antimicrobial, Effects of Rubia cordifolia Aqueous Root Extract

      Barlow, R.; Barnes, D. A.; Campbell, Anna M.; Nigam, Poonam S.; Owusu-Apenten, Richard K.; University of Chester, University of Ulster (Sciencedomain international, 2015-11-10)
      Aims: To evaluate the total antioxidant capacity (TAC) of Rubia cordifolia root extracts, to test anticancer activity against MDA-MB-231 breast cancer cell lines, and to evaluate antimicrobial activity of the same extract versus six Gram-positive and negative bacteria. Study Design: In vitro. Place of Study and Duration: School of Biomedical Sciences, Ulster University, July 2014-Sept 2015. Methodology: TAC was tested using ABTS, DPPH, FRAP and Folin assays and values were expressed as mg-gallic acid equivalents per 100 g (GAE/100 g) of sample. Anticancer properties were examined against MDA-MB-231 breast cancer cell lines using Sulforhodamine B assay. Antimicrobial activity was examined using a disk diffusion assay with three Gram-positive (Staphylococcus epidermidis, Staphylococcus aureus and Bacillus cereus) and three Gram-negative (Escherichia coli, Pseudomonas aeruginosa, Salmonella typhi) bacteria. Results: TAC of dry extracts of Rubia cordifolia ranged from 523±43 to 4513±208 (mg GAE mg/100 g) depending on the method of analysis, ABTS> FRAP> Folin > DPPH methods. R. cordifolia dry extract showed cytotoxicity against MDA-MB-231 with IC50 = 44 µg/ml or 5.1µM GAE. No antimicrobial activity was observed against the three Gram-positive, or three Gram-negative bacterial species using the water extract or R. cordifolia. Conclusion: R. cordifolia aqueous extract possess high total antioxidant capacity but values depend on the method of analysis. R. cordifolia extract inhibits MDA-MB-231 breast cancer cells proliferation but nil anti-bacterial activity was observed for three Gram-positive and three Gram-negative bacterial strains tested.
    • Application of immunological methods for the detection of species adulteration in dairy products

      Hurley, Ian P.; Ireland, H. Elyse; Coleman, Robert C.; Williams, John H. H.; University College Chester (Wiley, 2004-10-20)
      A number of enzyme-linked immunosorbent assays (ELISAs) have been developed for the detection of milk adulteration in dairy products. Target antigens have been caseins, lactoglobulins, immunoglobulins and other whey proteins. Polyclonal and monoclonal antibodies have been used in a variety of formats including direct, indirect, competitive and sandwich ELISAs. ELISAs have been successfully applied to the detection of cows' milk adulteration of sheep, goat and buffalo milk. Goat milk adulteration of sheep milk has also been detected. A number of ELISAs have also been applied to cheese. It is recommended that ELISA should be used in combination with PCR to ensure compliance with current legislation.
    • Approaching the problem of defining 'health' and 'disease' from the perspectives of evolutionary psychology and Darwinian medicine

      Lewis, Stephen J.; Chester College of Higher Education (2001-09)
      Concepts of 'health' and 'disease' are of fundamental importance to ethical considerations regarding medical provision. Yet the terms have no clearly agreed definitions. In fact, the difficulty of defining health has led to most attention being given to defining disease instead. Here, two schools of thought have arisen: the 'naturalist' which argues that disease is an objective entity in itself and the 'normativist' which gives emphasis to the subjective nature of disease experience differing between cultures and through history. Respectively, these two schools characterize quantitative (or functional) and qualitative (or evaluative) views of disease. Although both schools offer important insights, they are essentially at odds. This poster outlines an approach that seeks to find a basis for a meeting (if not a unification) of these schools by adopting ideas and approaches from evolutionary psychology and Darwinian medicine. From the perspective of reproductive fitness, the question of whether health and disease can be said to exist as biological entities is addressed and the idea that all that matters is reproductivity is considered. It is suggested that attitudes regarding certain biological entities, such as physical or physiological states, serve adaptive functions. The suggestion is then made that, although open to social and cultural influence, attitudes towards and qualitative definitions of health and disease also have biological bases. Thus, it may be argued that evaluative definitions of disease have functional (evolutionary) bases, thereby linking the naturalist and normativist schools of thought. Important in this linkage, however, is acceptance of ideas from evolutionary psychology. The only discipline that currently unites the study of health and disease with that of evolutionary biology (including evolutionary psychology) is Darwinian medicine. It is within this discipline that new theoretical and evidence-based understanding of 'health' and 'disease' is likely to prove fruitful – in particular, in giving 'health' appropriately weighted attention.
    • Arabinoxylans from rice bran and wheat immunomodulatory potentials: a review article

      Fadel, Abdulmannan; Plunkett, Andrew; Li, Weili; Ranneh, Yazan; Tessu Gyamfi, Vivian Elewosi; Salmon, Yasser; Nyaranga, Rosemarie Roma; Ashworth, Jason (Emerald, 2018-02-12)
    • Arabinoxylans: Bioactivities in Relation to Their Molecular Structure

      Li, Weili; Zhang, Zhengxiao; Smith, Christopher J.; University of Chester; Manchester Metropolitan University (Bentham Science, 2017-03-09)
      Arabinoxylans are a group of compounds with a basic structure consisting of a xylose backbone with arabinose side chains. Variations in structure occur as a result of variations in the xylose chain length, the ratio of arabinose to xylose and the introduction of alternative side-chains. This allows for an enormous potential range of structures. Arabinoxylans are major components of the cell walls of cereals. They have been reported to have numerous health benefits. This chapter presents a systematic description of the molecular features of arabinoxylans and relates these to the different extraction technologies used to obtain them. The proposal, that their immune modulation activity is related to their molecular weight and structure, is presented. Results demonstrating the effects of various arabinoxylans in various in vitro immunological tests are discussed.
    • Archibald Geikie: His influence on and support for the roles of female geologists

      Burek, Cynthia V.; University of Chester (Geological Society of London, 2019-06-19)
      This chapter explores the interaction between Archibald Geikie and female geologists in their many different roles and within the social context of his life time (1835-1924). The roles adopted by female geologists altered around 1875 due to a change in the educational and legal background. Geikie’s attitude to female fieldwork and research publications changes through time too. His life is divided up into 5 different stages according to his influence. Case studies of both single and married women are explored looking at the influence and interaction they had with Archibald Geikie. They include Maria Ogilvie Gordon, Catherine Raisin, Annie Greenly, Gertrude Elles, Ethel Skeat and Ethel Wood. Was one female role more acceptable to him than others? Geikie seems to accept most of the roles they undertook and he supported them wherever he could.
    • Are urological patients at increased risks of developing haemostatic complications following shock wave lithotripsy (SWL) for solitary unilateral kidney stones?

      Thomas-Wright, S. J.; Banwell, Joseph; Mushtaq, Sohail; Williams, R.; Abdulmajed, I.; Shergill, Iqbal; Hughes, Stephen F.; University of Chester; Wrexham Maelor Hospital (Elsevier, 2014-04-01)
      INTRODUCTION & OBJECTIVES: During the past two years there has been an increase in the number of patients undergoing shock wave lithotripsy (SWL) in Welsh hospitals (United Kingdom) for solitary unilateral kidney stones. Serious complications of SWL include haematuria, acute kidney injury and sepsis. Currently, there are no simple blood tests available, which can predict complications following SWL. Here we have tested the hypothesis that SWL will result in changes to haemostatic function, increase endothelial and haemostatic involvement postoperatively. MATERIAL & METHODS: In this pilot study, ten patients undergoing SWL for solitary unilateral kidney stones, were recruited (n=10). From patients (6 male and 4 female) aged between 31-70 years (mean=50 years), venous blood samples were collected pre-operatively (baseline), at 30 minutes, 120 minutes and 240 minutes postoperatively. Specific haemostatic biomarkers [D-dimer, von Willebrand Factor (vWF), Prothrombin time and sE-selectin] were measured. RESULTS: D-dimer and vWF concentrations were significantly increased from baseline at 240 minutes postoperatively (p=0.05 and <0.01 respectively). Prothrombin time and sE-selectin parameters were not significantly changed following SWL. CONCLUSIONS: The observed increase in D-dimer and vWF concentrations suggests that these markers would provide a more clinically relevant assessment of the extent of haemostatic involvement due to surgery. Analysis of such markers, have the potential to improve the detection of complications occurring postoperatively, such as haematuria and acute kidney injury.
    • Aspartame in conjunction with carbohydrate reduces insulin levels during endurance exercise

      Siegler, Jason; Howell, Keith; Vince, Rebecca; Bray, James W.; Towlson, Chris; Peart, Daniel; Mellor, Duane; Atkin, Stephen; University of Western Sydney ; University of York ; University of Hull ; University of Hull ; University of Hull ; University of Hull ; University of Chester ; University of York (01/08/2012)
      As most sport drinks contain some form of non-nutritive sweetener (e.g. aspartame), and with the variation in blood glucose regulation and insulin secretion reportedly associated with aspartame, a further understanding of the effects on insulin and blood glucose regulation during exercise is warranted. Therefore, the aim of this preliminary study was to profile the insulin and blood glucose responses in healthy individuals after aspartame and carbohydrate ingestion during rest and exercise. Each participant completed four trials under the same conditions (45 min rest + 60 min self-paced intense exercise) differing only in their fluid intake: 1) carbohydrate (2% maltodextrin and 5% sucrose (C)); 2) 0.04% aspartame with 2% maltodextrin and 5% sucrose (CA)); 3) water (W); and 4) aspartame (0.04% aspartame with 2% maltodextrin (A)). Insulin levels dropped significantly for CA versus C alone (43%) between pre-exercise and 30 min, while W and A insulin levels did not differ between these time points. Aspartame with carbohydrate significantly lowered insulin levels during exercise versus carbohydrate alone.
    • Assessing efficacy of cardiac rehabilitation exercise therapy in heart failure patients

      Leslie, Rosalind (University of Chester, 2015-10)
      Background: Exercise-based cardiac rehabilitation (CR) is considered routine practice for patients following an acute cardiac event or surgical intervention. Although there is a seemingly strong evidence base supporting it for patients with chronic heart failure (CHF), provision in the UK remains poor for this patient group. In addition, data for CHF patients reported in key CR reviews and meta-analyses are not a true representation of the UKs CHF population. The transferability of current evidence into actual practice settings in the UK therefore remains incongruous. Rationale and aims: Study outcomes have typically included an increase in VO2 peak/ VO2 max, a decrease in natriuretic peptides, improved left ventricular function and improved health related quality of life (QoL). Access to facilities and equipment, such as cardiopulmonary exercise testing equipment is limited in the UK for the majority of CR services thus an alternative means of assessment and exercise prescription is required. The recommended alternative for testing CHF patients is the six-minute walk test (6MWT); this requires a given space and a full practice test, the latter which adds to valuable clinical and staff time available. Methods: The first set of studies of this thesis therefore investigated two adapted assessment procedures for use with CHF patients: i. the use of a shorter practice walk test of two minutes vs six minutes prior to a 6MWT and ii. the use of the space saving Chester step test with an adapted lower step height protocol to accommodate the anticipated lower fitness in CHF (4-inch vs 6-inch). Having determined a more practical and efficient means of assessing exercise capacity in CHF patients, this thesis then used the 6MWT to evaluate the efficacy of a typically recommended 12-week programme (for the UK) of exercise-based rehabilitation. It was the aim of this PhD to also combine the use of the Chester step test with cardiopulmonary measures as a corresponding physiological outcome in a sub-sample of participants; however due to resource problems, only validation of the low-step protocol was possible. In the main intervention study, the efficacy of a 12-week course of supervised moderate intensity exercise in CHF patients (ejection fraction <44%, NYHA class II to III) was then evaluated. For purposes of evaluating safety and recovery of any acute myocardial stress induced by exercise in CHF, a sub-group study was performed to evaluate the influence of an acute exercise session on two-day post-exercise levels of circulating NT-proBNP. Results: In this current suite of studies, participants were more representative of the UK CHF population than typically reported in the current evidence. Their profile involved a median age of 76 ± 16 years (mean: 67 years and range: 30 to 84 years). 98% of whom were prescribed beta-blockers, 66% were diagnosed with atrial fibrillation and 98% had two or more co-morbidities. Study 1 (Chapter 3a) verified the efficacy of a two-minute practice walk in comparison to the recommended six-minute practice walk prior to performing a baseline 6MWT in patients with CHF. Study 2 (Chapter 3b) demonstrated that a 4-inch Chester step test is a reliable assessment when space is an issue, but the criterion validity of the actual oxygen costs at each stage compared with those estimated in healthy populations were significantly lower than recommended estimations from healthy populations. Study 3 (Chapter 4) revealed individual variability in the acute response of NT-proBNP release to exercise that is worthy of further study. However the NT-proBNP data overall did not suggest a need for ‘rest days’ between exercise training sessions. The main intervention study (Study 4, Chapter 5) demonstrated a significant improvement in 6MWT performance responses, compared with control, where an increased walking distance of 25 m (p < .0001) was coupled with a reduction in heart-rate-walking speed index (T1 16.3 ± 7.3 vs T2 15.3 ± 8.7 beats per 10 walked; p < .0001). Perceptually, patients were walking faster for the same rating of perceived exertion (RPE 12 to 13). This improved aerobic functioning coincided with an improved NYHA class (T1 2.3 ± .5 vs T2 1.8 ± .6; p < .0001); however there was no change in resting NT-proBNP levels after 12 weeks. Patients in the “control group” who then went on to be offered the same 12-week intervention achieved similar outcomes, but delaying their commencement of an exercise programme by 12 weeks negatively impacted on participation uptake. Key findings and conclusions: These results have demonstrated that exercise training in CHF can lead to an improvement in both physical and perceived functioning (NYHA class). In light of some previous studies showing decreases in BNP following an exercise programme and others like this one showing no change, further questions are raised about the effect of different types and doses of activity being offered to CHF patients and the responsiveness to training of different types of patients (disease severity and demographics). The nature of the cross-over design of this study revealed that delayed commencement of exercise negatively affects participation uptake by patients, which supports current UK standards in aiming for early referral to CR.
    • Assessing Risk Factors for Reproductive Failure and Associated Welfare Impacts in Elephants in European Zoos

      Hartley, Matt; University of Chester (European Association of Zoos and Aquaria, 2016-08-02)
      Reproductive failure in elephants is thought to be caused or influenced by a range of factors such as obesity, infectious disease, husbandry, facilities, stress, behaviour, maternal experience, herd size and social grouping. Due to the low reproductive activity of the small zoo elephant population, scientific study into the relative importance of these factors is limited. This study takes an epidemiological approach using risk analysis methodologies to collate information from expert opinion, data set analysis and a targeted questionnaire to identify and assess a range of physical, behavioural and husbandry based risk factors, which may affect reproductive success in elephants housed in European Zoos. Much of our knowledge on reproduction in zoo elephant populations originates from North America where there are significant differences in herd structure, management practices, climate and mean age. By combining multiple sources of evidence including a large survey of reproduction in the European elephant population and eliciting expert opinion from scientists, zoo managers, veterinarians and keepers working with European zoo elephants in a structured, transparent and scientifically recognised process it has been possible to identify the most important causes of reproductive failure and assess the influence of a range of potential confounding factors. Important causes of reproductive failure included lack of access to a compatible bull, herd stability and compatibility, allomothering or maternal experience, management practices at parturition and the impact of Elephant Endotheliotropic Herpes Virus. This work is to be used in the development of evidence-based elephant management and welfare recommendations and highlights priority areas for further research.