• Quality Control Systems of Gum Arabic in Sudan

      Al-Assaf, Saphwan; Eldigair, Hashim, Y. (University of Chester, 2018-11)
      Gum arabic is the oldest tree gum exudate, and has been in use since 4000 BC. Currently, it is used as an emulsifier (E414) in a number of processes, such as producing sweets and soft drinks. This study examines local practices such as tapping, collecting, transporting, sorting, and storing in various production areas in Sudan. Furthermore, it also investigates statistical differences in the key variables such as moisture content, optical rotation, ash content, viscosity, pH, colour Gardner index, colour Lovibond and tannin content in various production regions in Sudan during four seasons, namely 2013, 2014, 2015 and 2016. This study offers originality as the first investigation to combine labour practices related to quality control systems in a Sudanese context, using both primary and secondary data. Primary data were obtained from survey questionnaires (n=413 out of 800) distributed, giving a response rate of 52%, using chain-referral sampling among gum arabic farmers, managers, supervisors, and stakeholders. In addition to the questionnaires, open-ended (interview) questions were distributed to stakeholders and experts, by employing maximum variation sampling (n=15 out of 20 questions distributed, providing a response rate of 75%). Other primary data, namely, analytical and experimental data, were obtained from Nopec Quality Control Laboratory in Khartoum, and from the Hydrocolloids Research Centre at the University of Chester. Secondary data (national production) was obtained from the Sudan Customs Corporation via the Gum Arabic Board in Sudan. The findings of the survey questionnaires generally reveal that most workers tend to (i) work for relatively long time in the gum arabic industry, (ii) are knowledgeable about quality control systems, and (iii) aware about the best methods for maintaining product quality, collecting and storing gum arabic (r=-0.821). Specifically, the strongest correlation coefficient (p=0.001) were found between the worker’s age and the duration of working in the gum arabic industry (r=0.655). That is, the older respondents tend to be male while younger respondents tend to be female (r=-0.623). In addition, the majority of respondents (r=0.476) were knowledgeable about the production areas of gum arabic in Sudan as well as the location of the main auction market in Al Obeid. The findings from expert interviews indicate that there are both facilitative and hindrance factors that affect gum arabic development; these are related to infrastructure, technology, socio-economy, and relevant institution. The hindrance factors are, inter alia, the existence of relatively higher taxes, inefficient transportation, outdated technology and inconsistent quality control systems used by various gum arabic processing companies. Conversely, there are also facilitative factors such as financial assistance (the sheilla system) for farmers from banks, regular training, and methodical improvement of tapping through the use of modernised tools. The most significant factor is the agreement by all interviewees that better quality control systems should be a key to the development of this product therefore, allowing the suppliers to offer a quality product rather than a commodity. The results of secondary data reveal an increase in export trends from 2012 to 2018, indicating continuous growth in the industry and in particular for Acacia seyal compared to the previously held standard of Acacia Senegal. Finally, the findings of the analytical data reveal that key variables while differ across the various production season, the quality of the material from a given production area does not differ significantly. This is the major finding of this study whereby using reliable supply chain, traceability system and quality control measurements it would be possible to supply gum arabic with certain characteristics suitable for a given application. In conclusion, the findings are useful addition to our knowledge and potentially of commercial impact.
    • Development, Digestibility and Oxidation Properties of LC3PUFA Nanoemulsion and Its Effects on Sensory Profile of Food

      Zhou, QiQian (University of Chester, 2019-02)
      The long chain omega-3 polyunsaturated fatty acids (LC3PUFA) in human diets are mainly derived from oily fish and fish oil based supplements. Currently, the consumption of oily fish in the UK is far below the recommended level. LC3PUFA's non-fish sources such as algal oil with DHA (docosahexaenoic acid) are particularly important for vegetarians, non-fish eaters, and pregnant women. In previous work, high DHA vegetative algal oil load 50% w/w was successfully used to develop an oil-in-water nanoemulsion system suitable for functional food enrichment. The aims of this study included to investigate the effect of selected emulsifiers on oil-in-water nanoemulsions of algal oil prepared using ultrasonic technology. To improve the stability and digestibility of nanoemulsions within an In vitro digestion model. To examine the oxidation stability of nanoemulsions of algal oil and bulk algal oil with composition and droplet size changes during a 5 weeks storage trial at a temperature of 4 °C, 20 °C and 40 °C respectively. To evaluate sensory properties and consumer acceptability of food products with the incorporation of resulted nanoemulsion and find out possible relationship between the sensory profile of foods and the characteristics of added nanoemulsion. Nanoemulsion of LC3PUFA algal oil was developed with selected 6% w/w emulsifiers, including Lecithin (LN), Tween 40 (TN), Tween 60, equal ratio of Tween 40 and lecithin (LTN), 50% w/w Algal oil and 44%w/w water using a homogenizer and ultrasound processor. The results show that the nanoemulsion has been stabilised with selected emulsifiers (LN, TN & LTN) and the smallest droplet size of nanoemulsion was obtained using the combination of lecithin and Tween 40 at ratio 50:50. The In vitro digestion experiments were conducted with a model of fed state gastric and duodenal digestion using method of Lin et al (2014). The results show that the omega-3 oil nanoemulsion (LE/TW 50:50) were stable over 60 min in the gastric phase, in contrast omega-3 nanoemulsion (LE 100%) was destabilised at the gastric phase in 60 min, in which the droplet size diameter was significantly larger than at the beginning of gastric phase (P ≤ 0.05). The droplet size, fatty composition and oxidised compounds were measured to compare bulk algal oil and nanoemulsions stabilised with lecithin (LN) and Tween 40 (TN) solely and in combination (LTN) over a storage period of 5 weeks at temperatures of 4, 20 and 40°C. The results show the droplet size of nanoemulsions had no significant changes for samples stored at tested temperatures over 5 weeks storage. There were no significant differences in DHA composition within the weeks and temperatures used. For the GCHS analysed results, the increase in temperature to 40 ºC and storage time had a significant effect on the development of propanal for all samples (P≤0.05). Nanoemulsions prepared with lecithin alone had significantly higher development of propanal in week 1 at both 40 ºC and 20 ºC (P≤0.05). Lecithin (sole and combination with Tween 40) had more significant increases in oxidised volatiles at 40°C, which may be due to the instability of linoleic acid found in lecithin molecules which located in the outer layer of the oil droplets. There were no significant increase in oxidised compounds from the beginning to the end of storage for all tested samples stored at 4 °C. The sensory testing was also conducted on white sauce incorporated with omega-3 nanoemulsions with selected emulsifiers and bulk algal oil. The results show that the sensory attributes and overall acceptability of foods enriched with omega-3 nanoemulsion were statistically significantly lower than that of control sample (P≤ 0.05). Overall, the smallest droplet size of nanoemulsion was achieved with combination of lecithin and Tween 40 at a ratio of 50:50 by using ultrasonic processor. The stability and digestibility of nanoemulsion with the combination of lecithin and Tween 40 was improved in an In vitro digestion approach. A storage period of 5 weeks and temperature have no significant effect on the droplet size of tested nanoemulsion samples. However, there is a significant increase of the oxidised volatiles at 40 °C for all samples. Sensory testing show the white sauce with nanoemulsion has a stronger fishy taste and less overall liking than with bulk oil, indicating the smaller drop size is more ready to spread and reach the sensors of the mouth.
    • The interaction between the physical and mental loads associated with actual and simulated rugby league performance

      Highton, Jamie; Twist, Craig; Mullen, Thomas (University of Chester, 2019-09)
      The aim of the current thesis was to develop knowledge of the ‘loads’ associated with rugby league match-play, with a particular focus on the effects of altered mental loads before and during exercise indicative of a rugby league match. Chapter 3 examined the test-retest reliability of movement, physiological and perceptual measures during and after a novel rugby match simulation, where movement commands were more random than those typical of match simulations. The most reliable measure of external load during bouts of the simulation was relative distance (typical error [TE] and coefficient of variation [CV%] = 1.5-1.6 m.min-1 and 1.4-1.5%, respectively), with all other movement characteristics possessing a CV% <5%. The most reliable measure of internal load, neuromuscular function and perceptual measures were for %HRmax during bout 1 (TE and CV% = 1.4-1.7% and 1-4-2.1%, respectively), MVC before (TE and CV% = 10.8-14.8 N·m and 3.8-4.6%, respectively), and average RPE (TE and CV% = 0.5-0.8 AU and 3.6-5.5%, respectively). The conclusion of this chapter was that randomisation of the movements during simulated activity to better reflect intermittent team sports has no detrimental effect on its reliability. Studies can therefore confidently examine alterations in several perceptual, neuromuscular, physiological and movement load measures related to rugby activity using stochastic movements. Chapter 4 examined the responses to a simulated rugby league protocol that was designed to include more random commands, and therefore require greater vigilance, than traditional team sport simulation protocols. The randomised simulation (RDM) was matched for the number and types of activity performed every 5.45 min in a control trial (CON), but included no repeated cycles of activity. The RDM trial was more mentally demanding than CON (Effect size (ES) = 0.56; ±0.57). Self-paced mean sprint performance increased in RDM (22.5 ± 1.4 vs. 21.6 ± 1.6 km∙h-1; ES = 0.50; ±0.45), which was accompanied by a higher RPE (14.3 ± 1.0 vs. 13.0 ± 1.4; ES = 0.87; ±0.54) and a greater number of errors in the Stroop Test (10.3 ± 2.5 vs. 9.3 ± 1.4 errors; ES = 0.65; ±0.67). MVC peak torque (CON = -48.4 ± 31.6 N.m, RDM = -39.6 ± 36.6 N.m) and voluntary activation (CON = -8.3 ± 4.8%, RDM = -6.0 ± 4.1%) was similarly reduced in both trials. Providing more random commands, requiring greater vigilance, can therefore alter performance and associated physiological, perceptual and cognitive responses to team sport simulations. Chapter 5 describes the subjective task load of elite rugby league match play using the NASA-TLX and examines their association with several contextual match factors, technical ii performance and external movement demands. Linear mixed modelling revealed that various combinations of contextual factors, technical performance and movement demands were associated with subjective task load (NASA-TLX). Greater number of tackles (η2 = 0.18), errors (η2 = 0.15) decelerations (η2 = 0.12), increased sprint distance (η2 = 0.13), losing matches (η2 = 0.36) and increased perception of effort (η2 = 0.27) lead to most likely – very likely increases in subjective total workload. These data provide a greater understanding of the internal load and their association with several contextual factors, technical performance and external movement demands during rugby league competition. The purpose of the final empirical chapter (Chapter 6) was to describe the effects of mental fatigue on simulated rugby league performance and to determine the effects of caffeine supplementation on simulated rugby league performance in the presence of mental fatigue. Completing a mentally demanding task increases participants’ subjective rating of mental fatigue (pre = 29 ± 25 AU; post = 55 ± 20 AU) immediately before completing a simulation protocol. Impairments in sprint speed (ES = -0.18; ±0.19), sprint to contact speed (ES = -0.20; ±0.27), high-intensity running (ES = -0.30; ±0.24), high metabolic power > 20 W·kg-1 (ES =-0.50; ±0.51) and time to complete a passing accuracy task (ES = 0.54; ±0.63) were observed after mental fatigue. Caffeine supplementation (5 mg.kg-1) attenuated several adverse effects of mental fatigue before exercise replicating the demands of rugby league match play, with increased sprint speed (ES = 0.40; ±0.18), high-intensity running (ES = 0.50; ±0.53), high metabolic power > 20 W·kg-1 (ES = 0.33; ±0.38) and decreased time to complete a passing accuracy test (ES =-0.70; ±0.45). Mental fatigue affected internal loads, external loads and skill performance during simulated rugby league match play that appear to be centrally regulated by a decreased motivation and increased perception of effort. However, a single dose of caffeine taken 60 min before performance can attenuate several of these negative effects. In summary, the current thesis highlights several interactions between the physical and mental loads associated with actual and simulated rugby league performance.
    • A Geoconservation perspective on the trace fossil record associated with the end – Ordovician mass extinction and glaciation in the Welsh Basin

      Burek, Cynthia; Hosie, Lottie; Nicholls, Keith H. (University of Chester, 24/03/2019)
      In this thesis I have illustrated the value of our geological heritage and geodiversity by focussing on a particular detailed aspect of the geological and palaeontological record, i.e. the trace fossil record associated with the end Ordovician (Hirnantian) global glaciation and extinction episode. The major elements of this work that are new are: • a significantly improved understanding of the nature of the soft sediment deformation, and in particular the role of “debrites” as basal landslide decollements in the Lower Palaozic Llangrannog rock succession of West Wales, • a much more detailed description of the trace fossil ichnocoenose present in the Llangrannog succession than has previously been published • an improved understanding of the nature of the ecological perturbation associated with the Hirnantian (Late Ordovician) Glacial “ice-house”, and the apparent role of an opportunistic soft body fauna in filling ecological niches vacated as a consequence of the associated extinction. • Considerable thought has been given to the question of how to value abiotic nature, and it is argued that the methods of conservation valuation associated with “Geosystem services” and in particular “Natural Capital” hold considerable potential for the Geoconservation community to engage with the public and with policy makers. • As a direct result of this research, two formal proposals have been put forward for new RIGS sites, together with a new geological SSSI.
    • The effects of targeted therapy on cell viability and apoptosis on CML and AML cell lines

      Williams, John; Ireland, Elyse; Marsico, Paolo (University of Chester, 15/01/2019)
      Tyrosine kinase inhibitors (TKIs) are currently the first therapy option for chronic myeloid leukaemia (CML) and acute myeloid leukaemia (AML) patients. However, many patients affected by CML and AML may develop resistance to TKIs or may not recover under this treatment regime. New potential and more effective treatments are recently emerging. Heat shock protein inhibitors (HSPIs) and the proteasome inhibitor Bortezomib are drugs which have been yet to be successfully tested on leukemic patients, despite being successful on other malignancies such as multiple myeloma (MM). The combination between HSPIs and Bortezomib could potentially be successful in killing leukemic cells, by enhancing their respective molecular mechanisms. Indeed, HSPIs would bind to HSP72 avoiding the protein to exert its ligase function to the proteasome, whilst Bortezomib could stop the ubiquitinated proteins to enter the proteasome and ultimately inducing apoptosis. To test the effects of such combination, cell viability was measured via MTS assay, apoptosis levels were tested through Annexin V\PI assays. Involvement of HSP72 and pro-survival protein Bcl-2 were measured via flow-cytometry. The cells were administered with HSPIs and Bortezomib first as single agents for 24 hours, to establish working minimal concentration. Also, the drugs were tested for a shorter time, to understand when the drugs start to be effective. It emerged that one hour is sufficient for the drugs to give an initial effect in terms of cell viability and apoptosis. Following, combination experiments of HSPIs and Bortezomib were performed; the first drug was administered for one hour, the second following one hour and the cells were incubated for 24 hours. This was repeated alternatively for both type of drugs on the different cell lines. MTS and Annexin V\PI showed that there is not a synergistic effect between drugs, but instead there is antagonism. No necrosis was found at any level of the study. The cells were then probed for HSP72 and Bcl-2, to investigate their involvement in apoptosis mechanisms. Following 6 hours of combined and single agent treatment, both type of drugs inhibit HSP72 but failed to reduce the expression of Bcl-2, particularly on AML cells. It is thus proposed that CML and AML cells may die by apoptosis following a short time of treatment with HSPIs and Bortezomib by an extrinsic pathway of apoptosis, independent from Bcl-2 involvement and from mitochondrial pathway of apoptosis. This study may be the first to indicate a potential use of HSPIs and Bortezomib on CML and AML patients for a short time of treatment, although not in combination. Future studies are needed to further investigate the mechanisms of action of these drugs, aiming to potentially give CML and AML patients another successful therapy option to overcome resistance to canonic chemotherapy.
    • ‘Life in the Travelling Circus’: A Sociological Analysis of the Lives of Touring Professional Golfers

      Fry, John (University of Chester, 2014-08)
      As sports become more professionalised and international in scope athletes increasingly migrate from one country to another. These individuals are required to adjust and adapt quickly when moving internationally. Literature on sports migration, however, tends to focus on routes and pathways rather than the effects of movement on the athletes themselves. The aim of this study, therefore, was to explore how the frequent workplace circulation inherent in the lives of highly skilled migrants affects their social selves. Using professional golf as a case study, this project includes an analysis of family issues, relationships between players, pay and conditions, and technical approaches to playing golf. Interviews were conducted with 20 male professional golfers and analysed from a figurational standpoint. As golf tournaments are increasingly staged in a myriad of different countries players are required to spend longer periods of time away from home and experience intense feelings of loneliness and isolation. It is argued that golfers are not isolated in terms of people who they have around them while on tour, but rather in terms of lack of contact with people who they have positive meaningful feelings towards, such as their family and friends. To help reduce this loneliness, golfers develop behaviours that foster temporary we-group alliances with other players they perceive to be similar to themselves. People in such groups are friends, characterised by bonds of togetherness, while also enemies showing evidence of conflicts as they are in direct competition for a share of the overall prize money. Indeed the monetary rewards available for top golfers continues to increase, however, such recompense is only available to small numbers and the majority fare poorly. It is argued that the prize money breakdown fosters internalised behaviour constraints whereby many players ‘gamble’ on pursuing golf as their main source of income despite the odds against them. This habitus is strengthened given the significant financial investments many players have made to fulfil their childhood dreams, which further blurs their ability to see the reality of their lives. The result is many golfers are constrained to develop networks with sponsors for financial reasons which leaves some with conflicting choices between regular income, and adhering to restrictive contractual agreements, or the freedom to choose between different brands. As such, overall the results of this study highlight the importance of considering the cultural and social adaptations required in the life of a transient migrant.
    • A sociological analysis of the monetisation of social relations within the working lives of professional footballers

      Bloyce, Daniel; Law, Graeme C. (University of Chester, 22/11/2018)
      In recent years one of the most commonly discussed issues in professional sport, and in particular Association Football, has been the pay of professional athletes. However, much of this literature is largely based on assumptions, speculation or broad financial reports, with little, if any, focus on the potential impact on the athletes’ lives. Therefore, the aim of this research was to examine the role money plays in the relationships within the working lives of professional footballers. Using professional football as a case study, this project examined a number of key areas: the consumption of products by footballers as a demonstration of economic power and wealth in an environment where wages are a taboo subject, the complex nature of contract negotiations and the impact this can have on relationships within their working lives. In addition to these areas, the thesis examined how money is used as punishment for players to try to encourage them to conform to the expected codes of behaviour set by club managers and officials, and ways in which players attempt to break their highly routinised daily life. Semi-structured interviews were conducted with 34 male professional footballers and analysed using concepts from the sociology of money. It is argued that image has become an important factor for many professional footballers. Displaying wealth through ‘conspicuous consumption’ was also important in an environment where wages are a secretive subject, as it is suggested that the ‘more you have, the better you are’ and therefore some players even felt that this would impact on the way in which they were valued by the club hierarchy (as well as their teammates within the club). Value was also important through contract negotiations, as the more a player was valued by a club, the greater balance of power they had within the negotiation process. It is argued the negotiation process has become more complex since the introduction of the Premier League, as more people are typically involved. It was also evident that money was a major factor for players when deciding on contracts or having to relocate, which led to feelings of loneliness for some players and their families. Players are heavily regulated and constrained within their lives, one-way players are constrained, by the club officials, is through financial punishment. Players discussed several methods of trying to break the routinisation that such constraints introduce. One of those was gambling. It is argued that some players, due to the technological advances, were able to gamble in a covert manner and keep their gambling losses private, which can impact on the performance, health and wellbeing of the players. Overall the results of this study highlight the increasing monetisation of social relationships within professional football and that such trends are significantly impacting on the relationships within the working lives of professional footballers.
    • The Use of Qualitative Risk Analysis Methods to Facilitate Decision Making in the Management of Health and Welfare in Wildlife

      Hill, Sonya; Smith, Tessa; Hartley, Matthew (University of Chester, 08/10/2018)
      This thesis is composed of a series of papers, all of which have been published in peer reviewed publications. The papers use the recognised process of qualitative risk assessment in a range of scenarios in the field of wildlife health and welfare in both in situ and ex situ environments. Chapter 1 discusses the challenges faced regarding availability of empirical data in field of wildlife and zoological health and welfare and justifies the exploration of techniques to assist with decision making. The development of risk analysis and its integration with risk management and risk communication to become risk assessment is described before being put into the specific context of wildlife and zoological disease. Chapters 2 and 3 consider two scenarios where disease risk assessment is well established as a tool, importation across national borders and in conservation interventions. Chapter 2 develops the standard import risk assessment approach to include multiple species and multiple diseases. Chapter 3 reviews developments made over the last 25 years and proposes best practice approaches to implement. Chapter 4 describes how the risk assessments formulated as described in Chapter 3 are used for licensing purposes emphasising the importance of risk management and communication. This theme is continued in Chapter 6 where the integration of risk assessment and evidence based decision making is considered in the broad context of a strategic approach to wildlife health bringing together the outcomes and processes described in Chapters 2, 3, 4 and 5. The papers in Chapters 2,5 and 8 are focused on how risk analysis aids in development of disease control approaches and policy. The evidence base is composed primarily of peer-reviewed literature supported by expert review of the finalised assessment. Chapter 7 uses risk assessment in an applied scenario, taking the recognised process and modifying it to structure an active disease investigation demonstrating the versatility of the technique. Chapter 9 takes this a step further by again adapting the methodology which, has historically been used primarily for infectious diseases, to consider reproduction and assess risks to welfare rather than purely health. The paper in Chapter 9 builds on the methodology by combining existing peer-reviewed literature with data collected specifically for the purpose of feeding into the assessment and utilising a stakeholder and expert opinion elicitation workshop to obtain data too. These process are proposed and described in Chapter 3. The final chapter critically reviews risk assessment, highlighting three key areas of potential weakness and proposing approaches to address these criticisms. The value of the approach in wildlife and zoological health and welfare as demonstrated by this series of papers is described
    • The role of MAPK signalling pathways in leukemic cell death

      Williams, John; Ireland, Elyse; Cordingley, Michelle (University of Chester, 18/09/2018)
      Mitogen-activated protein kinase (MAPK) signalling pathways are important signalling pathways involved in mediating various cellular processes including both cell survival and cell death. The c-Jun N-terminal kinase (JNK) pathway, the p38 pathway and the extracellular signal-regulated kinase (ERK1/2) pathway are three well-studied conventional MAPK signalling pathways. Previous research has shown these MAPK signalling pathways play an important role in the development and progression of leukaemia and in the response of leukemic cells to therapy. Whilst it appears to be well established that the constitutive activation of ERK mediates leukemic cell survival, the roles of the JNK and p38 signalling pathways in leukemogenesis, in particular the role in acute myeloid leukaemia (AML), are less well understood. This thesis investigates the role of the JNK, p38 and ERK signalling pathways in leukemic death. MAPK signalling pathways were targeted in the U937 monocytic cell line using small molecule MAPK inhibitors in combination with various cell stressors: UV light, chemotherapeutic agents (doxorubicin and vincristine) and heat treatment. The effects on cell death were examined using plate-based assays, flow cytometry and fluorescence microscopy. Preliminary investigations were also performed in peripheral blood mononuclear cells (PBMCs) from healthy individuals to allow a comparison to non-leukemic cells. Results show inhibition of ERK signalling in U937 cells induced cell death and ERK signalling had little effect on UV-induced and heat treatment-induced cell death. JNK signalling and p38 signalling provided protection against UVinduced cell death in both U937 cells and in PBMCs from healthy individuals. JNK and p38 signalling mediated cell survival in response to heat treatment to a certain extent. JNK signalling was required for the induction of cell death induced by doxorubicin whereas p38 signalling provided a level of protection against doxorubicin-induced cell death. U937 cells were found to be more sensitive to vincristine treatment than PBMCs from healthy individuals and the activation of JNK and p38 signalling was essential for vincristine-induced cell death in U937 cells. Taken together, the results presented in this thesis demonstrate that the roles of the JNK, p38 and ERK signalling pathways in leukemic cell death are stimuli-specific. This highlights the importance of understanding the involvement of particular pathways in the response to specific chemotherapeutic agents, in order to provide effective leukaemia therapy. Therapeutic inhibition of MAPK signalling pathways to increase the sensitivity of leukemic cells to chemotherapy could be beneficial when MAPKs are involved in providing protection against chemotherapy-induced cell death. For chemotherapies which require MAPK activation for cell death, failure to activate MAPKs may provide a mechanism for chemoresistance. Therapeutic methods to enhance activation of the pathways provide a possible approach to increase the susceptibility of leukemic cells to death.
    • Manipulation of apoptosis in cancer cells

      Williams, John H. H.; Ireland, Elyse; Sahib, Muneera M (University of Chester, 18/09/2018)
      Conventional cancer therapies can have severe side effects, so new strategies to limit these needs to be investigated. Several anticancer agents induce the expression of tumour suppressor gene p21 in colorectal cancer cell line HT-29. Interestingly, the stress protein HSPA1A is also often elevated in tumour cells and has an anti - apoptotic activity. The main aim of this study was to examine whether a two - pronged approach, overexpressing p21 (using genetic approach and inhibition of HSPA1A using pifithrin - µ would be effective in inducing apoptosis in tumour cells. Chitosan or BSA based delivery systems were evaluated for cytotoxicity, with the intension of using it for plasmid DNA based cell transfections in this study. The interaction of HSPA1A protein in combination treatments involving UV radiation and hyperthermia at 42℃ were also evaluated to perceive the various roles of HSPA1A in arresting colorectal cancer cells. Colorectal cancer cell lines HT-29 and leukaemia cancer cell lines U937 were used in the study. All experiments were performed with cancer cell lines maintained in culture medium devoid of antibiotics. Cell cytotoxicity were evaluated using MTS and PI assays. The rate of apoptosis was determined using annexin V and PI staining by flow cytometry. Chitosan or BSA based microparticles or microgels were observed for size determination or morphology using scanning electron microscopy. Full length human p21 inserted plasmid DNA was a gift from Mien - Chie Hung, Addgene, USA. HT- 29 cells were subjected to p21 plasmid DNA transfection effects. Cells were treated with pifithrin - µ (15µM) prior to gene transfection to address its combined effect with p21 plasmid DNA transfection. HSPA1A and p21 protein expression studies were analysed using FITC labelled antibodies by flow cytometer. Combination studies with HSPA1A inhibitor pifithrin - µ and UV reflected enhanced cytotoxicity compared with either of the treatments independently. Hyperthermia at 42℃ induced apoptosis by MTS assay, which was confirmed by flow cytometric analysis in both the cell lines tested. Considering the cytotoxicity reflected by the chitosan or BSA delivery systems in drug free states, the p21 plasmid DNA transfection was carried out using lipofectamine 2000. Both overexpression of p21 and inhibition of HSPA1A protein with pifithrin - µ enhanced the rate of apoptosis with statistical significance of (p-<0.0001****) compared to the respected controls. The data in this thesis suggests the inhibition of HSPA1A in combination with increased p21 would be a promising therapeutic strategy for the treatment of colorectal cancers.
    • Assessment of strength and power responses to resistance exercise in young and middle-aged trained males

      Twist, Craig; Lamb, Kevin; Fernandes, John (University of Chester, 31/08/2018)
      Little is known about the muscle function capabilities of trained middle-aged males and how they differ to younger counterparts. Accordingly, the overall aim of the research documented in this thesis was to compare the acute muscle function responses to resistance exercise in middle-aged and young resistance trained males. The first study (Chapter 3) examined the intra- and inter-day reliability of an ecologically valid device (FitroDyne rotary encoder) for measuring upper and lower-body muscle function during three popular multi-jointed resistance training exercises (bench press, squat, and bent-over-row), and confirmed that it was capable of detecting moderate changes in muscle function across a range of submaximal loads. In the second study (Chapter 4) the load-velocity and load-power relationships were investigated during the same exercises among 20 young (age 21.0 ± 1.6 y) and 20 middle-aged (age 42.6 ± 6.7 y) resistance trained males, and it emerged that, despite their regular training, the middle-aged males were unable to achieve velocities at low external loads and peak powers at all external loads as high as the young males across a range of external resistances. Study three (Chapter 5) proceeded to compare the internal (heart rate (HR), OMNI-ratings of perceived exertion (RPE) and sRPE) and external (peak velocity and power and volume load) loads experienced during high volume squatting exercise, and the fatigue responses among nine young (age 22.3 ± 1.7 years) and nine middle-aged (age 39.9 ± 6.2 years) resistance trained males. The findings highlighted that internal, but not certain markers of external (peak power and volume load), load responses can be monitored during exercise in a like manner between these age groups. Moreover, compared to young resistance trained males, middle-aged males can expect greater decrements in peak power after lower-limb resistance exercise. In the final study (Chapter 6), the time-course of recovery in nine trained young (age 22.3 ± 1.7 years) and nine trained (39.9 ± 6.2 years) and nine untrained (44.4 ± 6.3 years) middle-aged males after high volume lower-body resistance (muscle damaging) exercise was investigated. Of practical importance, it emerged that compared to the young males, the trained middle-aged males experienced greater symptoms of muscle damage and an impaired recovery profile, the implication of which is the need for trained middle-aged males to adopt strategies to enhance their recovery. Furthermore, both middle-aged groups experienced similar symptoms of muscle-damage, albeit the untrained group demonstrated greater losses in peak power at low and high external loads. For the first time, the current research has determined that middle-aged males, despite regular resistance training, are subject to losses in peak velocity and power output across a range external loads, compared to young males. When undergoing lower-body resistance training to ameliorate these decrements, applied practitioners can use internal load markers and peak velocity, but not peak power or volume load, to monitor trained young and middle-aged males alike. Furthermore, the muscle damage response (24 to 72 h), and losses in peak power (0 to 72 h), after lower-body resistance exercise are greater in trained middle-aged than young males. Consequently, future research should seek to corroborate these observations in upper-body exercise and determine the effectiveness of strategies (e.g. nutritional intake) to enhance recovery in middle-aged males.
    • Quantification of physical contact and its influence on simulated performance and recovery in rugby players.

      Twist, Craig; Norris, Jonathan (University of Chester, 07/08/2018)
      The aim of this thesis was to investigate the influence of physical collisions on internal (physiological and perceptual) and external (locomotive and accelerometer) load during simulated rugby league performance and fatigue responses in the days after. Chapter 4 examined the influence of physical contact type on internal and external load using a traditional soft tackle bag and custom-built tackle sled. Using a traditional tackle bag to simulate physical collisions resulted in likely faster sprint to contact speed (16.1 ± 1.5 c.f. 14.8 ± 1.1 km.h -1 ) but possibly lower overall high-speed running distance (27.7 ± 2.4 c.f. 28.4 ± 2.6 m.min-1 ). Also, the heavier tackle sled likely increased time at 91-100% HRpeak (12:58 ± 13:21 c.f. 6:44 ± 8:06 min:s) and resulted in greater lower limb fatigue reflected by the likely larger decrease in countermovement jump (CMJ) performance (5.9 ± 4.9 c.f. 2.6 ± 5.4%). Also of note was the variation in number of tackles detected using the automatic tackle detection feature compared to the actual number in the match simulation. During the Bag and Sled simulations ~53 and ~59 tackles were detected compared to 48 performed. The purpose of Chapter 5 was to investigate the influence of sprint to contact speed and contact type on automatic tackle detection using microtechnology. Repetitions were divided into three speed categories; walking, jogging and striding (1, 2.5 and 4 m.s -1 ) and four conditions: i) no contact standing upright (NCST), ii) no contact dropping to the ground in a prone position (NCGR), iii) contact with the tackle bag and remaining upright (CST), iv) contact with the tackle bag and going to ground (CGR). Similar tackle detection accuracy was observed between NCGR and CST conditions with one tackle observed in 41 and 43% of trials, respectively. While CGR resulted in the greatest frequency of correct tackle detection (62%), during 16% of trials two tackles were detected. During NCST, there were no tackles detected and 100% accuracy. The PlayerLoadTM results demonstrated that the metric can detect differences in movement speed, the inclusion of physical contact and changes in orientation during short periods of activity (8-10 s). In Chapter 6 the rugby league movement simulation protocol for interchange players (RLMSP-i) was modified to include a tackle shield collision to investigate the reliability of PlayerLoadTM metrics to quantify collision load. The coefficient of variation (%CV) for locomotive metrics ranged from 1.3 to 14.4%, with greatest variability observed for high-speed running distance (8.0 and 14.4% for Bouts 1 and 2, respectively). Accelerometer metrics CV% were 4.4 to 10.0%, while internal load markers were 4.8 to 13.7%. All variables presented a CV% less than the calculated moderate change during one or both bouts of the match simulation except from high-speed distance (m.min-1 ), %HRpeak and RPE (AU). The aim of Chapter 7 was to investigate the influence of contact type on external load metrics including PlayerLoadTM derivatives whilst controlling for total running distance. Participants were randomly assigned to one group to complete the match simulation with either a tackle shield (n = 10), tackle bag (n = 7) or no-contact (n = 10). Total PlayerLoadTM, PlayerLoadTM 2D (AU), PlayerLoadTM slow (AU) and PlayerLoadTM slow-ratio (%) were analysed from the accelerometer in addition to high- and low-speed running and sprint speed. Total PlayerLoadTM was likely lower for the Bag group compared to the Run group (498 c.f. 460 AU), with no clear differences between the other groups. 3 PlayerLoadTM slow for the Shield group (167 ± 26 AU) was very likely greater than both the Bag (133 ± 11 AU) and Run groups (128 ± 20 AU) but no clear difference was observed between the Bag and Run groups. No differences were observed in PlayerLoadTM 2D between any groups. High-speed running distance was likely lower in the Shield group (1056 ± 225 m) compared to the Bag group (1326 ± 245 m) and very likely lower compared to the Run group (1318 ± 175 m). Total PlayerLoadTM is not sensitive to contact type during simulated rugby league activity but does reflect greater high-speed running distance during a rugby league match simulation. However, PlayerLoadTM slow can detect the types of contact and might be preferred for quantifying match and training loads associated with physical contact. The purpose of the final empirical chapter (Chapter 8) was to determine the influence of contact type on in neuromuscular, perceptual and biochemical parameters associated with exercise-induced muscle damage. The participants were again assigned to one of three groups to complete the match simulation with a tackle shield (n = 6), tackle bag (n = 7) or no contact (n = 7). In addition to internal and external load measured during the match simulation, venous blood, muscle function and soreness measures were collected immediately (+0), +24 and +72 hours after the match simulation. Upper body neuromuscular performance and knee flexion torque likely decreased in the Shield group +0 and +72 hours after the simulation compared to the other groups while CMJ power likely decreased more in the Run group. All three groups demonstrated a very likely increase in IL-6 and IL-10 concentration immediately after the match simulation, but differences between the groups were unclear and values returned to baseline +24 hours after the simulation. In conclusion, current automatic tackle detection metric should be used with caution, particularly in training sessions where physical contact is replicated. Instead PlayerLoadTM and associated derivatives from the embedded accelerometer can provide a useful measure of contact-specific load during training and competitive matches. Physical contact type affected external load by modifying a participant’s running strategy during simulated match performance, thereby influencing site-specific fatigue during and after a simulated rugby league match. However, regardless of contact type, large increases in cytokine and leukocyte concentration are apparent with a return to basal values 24 hours after. Therefore it is not recommended to use such biomarkers in applied settings to quantify the magnitude of muscle damage specifically associated with physical contact.
    • Musculoskeletal Disorders in the Workplace: An examination of the underlying causes and contributory risk factors

      Fallows, Stephen; Gellatly, Pamela (University of Chester, 16/05/2018)
      Introduction The incidence of musculoskeletal disorders remain the most common single condition, by incidence, affecting the working population. This remains true even though the apparent historic causation of manual handling, has reduced significantly. Back pain alone has been termed a 20thcentury medical disaster, which has reverberated into the 21stcentury, with 85% of low back pain having no clear clinical diagnosis yet individuals continue to seek a clinical solution. Understanding pain remains as complex as ever with very little evidence to suggest progress. The overall scale and cost of MSDs in the workplace are not easily identifiable as objective and accurate data are rare. Other workplace incidence and costs are either, not recorded or not published, in documents or grey literature, that are generally only accessible to individual organisations on a regular basis. Objectives The epistemology of this thesis is complexity and the extent to which this influences outcomes. The trilogy of complexity considered includes: 1. The issues facing organisations in how they prevent and manage MSDs; 2. The individual’s perspective and what they understand about possible causation, their beliefs, fears and expectations; 3. The interface with clinical and non-clinical practitioners, and whether interventions provided, are beneficial to the individual. Consideration of the multiple perspectives that arise from the various influences affecting the organisation, the employees within that organisation and the practitioners, has been possible by the metaphoric use of a “bricolage” methodology, and suggests that the current medical model is no longer appropriate. Methods A mixed method research design comprising four studies was undertaken. Firstly, a retrospective quantitative study of data (n = 21,092) from benefits provided by four organisations followed by a qualitative case example study (n = 21) of supporting documents and clinical information. These studies then informed the need for a qualitative study (n= 9) symptomatic individuals who participated in a focus group and (n= 6) face-to-face interviews and finally a qualitative study of practitioners involved in the provision of treatment services to the participating organisations. The data from each study informed the others and the data merged with the findings from the literature review and common interventions. Conclusions A disparity was found between what has been identified in literature and what actually is considered in clinical practice. The healthcare industry operates in “silos” and this separation of disciplines is reflected in organisational management. The range of underlying risk factors, evident in modern society, which are affecting or may affect an individual’s future musculoskeletal health are not being addressed by the medical model, and practitioners require training, or need to work in a multidisciplinary team, if they are to improve long-term outcomes. This thesis discusses the complexity of the multifactorial nature of musculoskeletal health, and provides a framework to challenge current practice and promote a fundamental change in the way in we assess, and treat the range of MSDs including a move towards educating individuals to take personal responsibility.
    • Factors affecting the quality of Acacia senegal gums

      Al-Assaf, Saphwan; Hamouda, Yasir (University of Chester, 2017-04)
      Gum arabic is a natural gummy exudate from acacia trees and exhibits natural built-in variations commonly associated with hydrocolloids. This study is concerned with the determination of factors which could influence its properties and functionality. These factors include origin (location, soil type, rainfall), different collections, age of the trees and storage condition. Previous studies acknowledged the influence of some of these factors but somehow lack providing definitive answers to questions being asked by the end user and required for the development of Gum arabic industry in Sudan. Local knowledge as well as the various stages of gum collection and processing were reviewed in order to provide a clear background and the justification for the experimental design. In this study samples were collected from six plantations located in the west and east regions in Sudan. Samples were collected from trees of different age (5, 10, 15 and 20 years old) and also from different picking interval (1-4). Each sample was divided into three portions (UK, Khartoum and Port Sudan) and stored for 5 years in order to determine the effect of the respective location. Various analytical parameters (% loss on drying, Optical rotation, % protein, intrinsic viscosity, molecular weight and molecular weight distribution) were measured to fully characterise the gum samples and to determine their functionality (emulsification). The results obtained for all samples were consistent with those previously reported in the literature (see Chapter 4). The only exception, identified in a number of samples from the western region, is the high proportion (~30%) of high molecular weight fraction termed arabinogalactan-protein complex (AGP). The results clearly demonstrated significant variations between plantations located in western region compared with the eastern region. However, the variations between the plantations within the same region are statistically not significant. High values of % protein, viscosity, Mw and % AGP were obtained from the 1st pick, from both regions, and then significantly decreased thereafter to the fourth pick. Samples from west region in Sudan, from 1st and 2nd pick and from tree age (15) years gave the highest viscosity, molecular weight, % AGP and superior emulsification performance compared to other samples from different tree ages. The regression statistical analysis for the physiochemical properties correlation with emulsification performance demonstrated the role of % AGP to be the most influential factor followed by viscosity. The major finding of this study is the effect of storage condition on the properties and functionality of Acacia senegal. An increase in the molecular weight for all stored samples (for 5 years) irrespective of region was evident and statically significant. However, this increase was more prominent for samples from the western region compared to the eastern region. The AGP fraction was increased by the storage treatment up to 40% in Port-Sudan, 20% in Khartoum-Sudan and 15% in UK. The result clearly demonstrated that the temperature and humidity are the crucial factors to induce the natural maturation process in acacia gums. Statistical analysis (linear regression) suggested statistically significant models and equations to predict and explain the variations in the physiochemical and functional properties based on the environmental factors, picking set and age of the tree.
    • The genetics and evolution of the critically endangered Trinidad Piping Guan Pipile pipile, synonym Aburria pipile.

      McDowall, Ian; Hosie, Charlotte A.; Grass, Amelia (University of Chester, 2018-02)
      The Trinidad Piping Guan, Pipile pipile synonym Aburria pipile (Jaquin, 1784) is the only endemic Cracid on the island of Trinidad. The species is currently listed as Critically Endangered and is considered to be in ‘on-going decline’ by the IUCN, BirdLife International and Cracid Specialist Group. This study aims to examine aspects of genetic variation and the evolution of the mitochondrial genome in the Trinidad Piping Guan utilising, for the first time, samples collected from individuals in the wild and reference specimens of the genus Pipile sourced from museum collections. In this study the complete mitochondrial genome of the Trinidad Piping Guan was sequenced for the first time. Analysis of intra-specific variation of wild Trinidad Piping Guan individuals using single nucleotide polymorphisms demonstrates extremely limited variation within the genes of the mitochondrial genome and nuclear gene intron sequences. Limited variation within this population is consistent with both historical and contemporary contractions of populations within a restricted island system, which may have serious implications for the future of this species in terms of both genetic diversity and conservation management. Phylogenetic analysis of the complete mitochondrial genome of the Trinidad Piping Guan enabled placement of the genus Pipile within the Galliforme evolutionary tree for the first time, and subsequently places the genus within the broader context of the Aves class. Mito-genomic analysis confirms that the Cracids are one of the basal Galliforme clades, and sister taxa to the Megapodidae. Phylogenetic placement of the Pipile genus is basal to that of the Crax species within the Cracidae family, indicative of an earlier evolutionary origin of the Piping Guans. The inclusion of the Trinidad Piping Guan, in the avian evolutionary tree using the whole mitochondrial genomes expands the current genetic phylogeny of the Cracid family, yielding a better understanding of evolutionary relationships among the Galliforme order and the diversification of modern avian lineages. This study has established novel molecular techniques for the analysis of mitochondrial DNA in historical specimens of the genus Pipile from museum reference collections. The analysis of inter-specific relationships within the genus Pipile has clarified the evolutionary and biogeographic relationships between the Piping Guan species. Additionally, the Trinidad Piping Guan is genetically defined for the first time as an evolutionarily significant unit, which represents a unique evolutionary pathway within this important genus in a closed island system on the island of Trinidad.
    • Exploring hygiene compliance in the small independent restaurant sector in Abu Dhabi

      Fallows, Stephen; Bonwick, Graham A.; Idriss, Johaina (University of Chester, 2017-10)
      Introduction: Food safety is widely recognised as one of the problems in the fight for improving public health. Many governments are trying to improve public health through reducing foodborne illnesses and setting the climate for implementing HACCP-based food safety management systems (FSMS). Following the global trend, Abu Dhabi Food Control Authority (ADFCA) launched the HACCP for Catering Project (2010 – 2014), which aimed at helping foodservice businesses, licensed in the Emirate of Abu Dhabi, in implementing HACCP-based FSMS. Purpose: The project team recognised the limited resources and the diversity in education levels, ethnic backgrounds, and number of languages spoken among managers/supervisors and food handlers, as points of concern in the small independent restaurant (SIR) subsector. Thus, the Salamt Zadna (SZ) initiative, a simplified FSMS, was developed to train SIRs on implementing a set of safe operating procedures to improve compliance with food safety laws and regulations. Previous studies in the GCC region have mainly focused on governments’ attempts to enhance public health by developing laws, regulations, and policies, and recounting the barriers to implementing food safety controls. Methodology: This thesis took a different approach to food safety issues in the GCC region. It is comprised of two studies, which were conducted in two groups of SIRs – seven SZ participants and five non-participants – licensed in Al Ain, a major city in the Emirate of Abu Dhabi. The first explored awareness and understanding of food safety, related laws, regulations, and policies, and attitudes towards ADFCA services and inspectors, among managers/supervisors, by interviewing them. The second examined the efficacy of SZ in improving food handlers’ food-safety behaviours by observing their conduct, and comparing between the two SIR groups. Results: The study indicated low levels of awareness and understanding of food safety, related laws, regulations, and policies, in both groups of managers/supervisors; regardless of whether or not they were SZ Cparticipants. Both groups of interviewees expressed both negative and positive attitudes towards ADFCA’s services and inspectors; sometimes by the same interviewees, within the same, or between the two groups. However, SZ participating SIRs were slightly more positive than their counterparts. Key results highlighted the low impact of SZ on changing food handlers’ behaviours, except in two areas; namely, the food handlers working in SZ-participating SIRs scored higher than the other group in handwashing and changing gloves between handling raw meats and other foods. Implications: This research adds a new dimension to the food safety profile of the UAE, since it is the first of its kind in the UAE and the region as a whole. Its originality opens the door for other researchers to increase the volume of research in this field, which would help in understanding and tackling the barriers to improving the food safety status in the country, as well as the region.
    • Learning to Play: How working-class lads negotiate working-class physical education

      Green, Ken; Scattergood, Andrew J. (University of Chester, 2017)
      Adults from the middle-classes are up to three times more likely to be regularly involved in sport than those from the working-class. The reason for this participation anomaly has been consistently linked to the differing lifestyles and opportunities to which young people from working and middle-class backgrounds are exposed. More specifically, working-class children are more likely to develop narrow, class-related leisure profiles and sporting repertoires during their childhood that serve to limit the likelihood of them remaining physically active in adulthood. In relation to this, one of the key aims of physical education (PE) in mainstream schools is to develop the range of skills and knowledge for all pupils and widen their sporting repertoires in an attempt to promote long-term participation throughout their lives. However, not only has PE provision in British mainstream schools been shown to be unsuccessful in promoting working-class pupils’ sporting/ability development, some suggest that the subject may even be perpetuating the social difference that has been shown to exist in relation to sports participation between social class groups. In order to address these issues the study set out to examine the extent to which the wider social background of white, working-class ‘lads’ and the actions and attitudes of their PE teachers came to impact on the way the lads influenced and experienced their PE curriculum/lessons. It also aimed to examine the impact that school PE then had on their sporting repertoires and participation in sport/active leisure outside of school. A total of 24 days were spent in Ayrefield Community School (ACS), a purposively selected, working-class state secondary school as part of a case study design. Over 60 practical PE lessons were observed that led to differing roles being adopted and guided conversations being conducted before, during, and after these lessons. Eight focus group interviews were also conducted with specifically chosen lads as well as one with the four members of male PE staff. Additional observations were also carried out during off-site trips, external visits, and in a range of classroom-based lessons. The findings were then considered and examined in relation to the work of the sociologists Norbert Elias and Pierre Bourdieu. The findings revealed that the pressures related to the modern education system and the social expectations linked to their working-class backgrounds caused a split between the lads at ACS in to three broad groups, namely: Problematics, Participants and Performers. These groupings came to impact on the ways that these lads engaged and achieved in school as well as the ways in which they came to negotiate and experience PE. The ‘Problematic’ group held largely negative views of education, but valued PE, especially when playing football, the ‘Participants’ were relatively successful at school yet apathetic regarding the content and delivery of their PE lessons, and a Performer group of lads emerged who engaged and achieved highly at school and participated in a range of activities in PE, but showed little intention of participating outside of school due to their pragmatic attitude to ‘learning’ in PE. Despite these differing school and PE experiences between the lads’ groups, the potential and actual impact of school PE on their sporting repertoires, skills, and interests was ultimately constrained by a range of issues. In the first instance the lads’ narrow, class-related leisure profiles and sporting repertoires linked closely to recreational participation with friends, alongside a lack of proactive parenting were significant limiting factors. In addition, the ability of some lads to constrain the actions of PE staff and peers to get what they wanted in PE rather than what they needed, and the negative views of most lads to skill development and structured PE lessons meant that PE at ACS was never likely to have a positive impact on the sporting repertoires and participation types/levels of its male pupils either currently or in their future lives.
    • Production of DNA aptamers with specificity for bacterial food pathogens

      Bonwick, Graham A.; Drasbek, Mette R.; Young, Niall; Smith, Christopher; McDowall, Ian; Kärkkäinen, Riikka M. (University of Chester, 2012-09)
      Aptamers are biomolecular ligands composed of nucleic acids. They can be selected to bind specifically to a range of target molecules and subsequently exploited in a fashion analogous to more traditional biomolecules such as antibodies. In this study a method for selecting new aptamers which specifically bind whole live bacterial cells is described. A non-pathogenic strain of Escherichia coli K12 was used to develop the method. A DNA library containing 100 bases long random nucleotide sequences was produced and the aptamer selection process was repeated nine times. An enzyme-linked technique was first used to detect bound aptamers thereafter fluorimetry and fluorescence microscopy methods were used for the detection. The aptamers were cloned and sequenced and the cloned aptamers produced with fluorescent labels. The E. coli K12-binding aptamers were used to demonstrate the detection of the bacterial cells in a complex food matrix, namely probiotic yogurt, by using fluorescence based detection method. The aptamer selection method with some modifications was also used to select aptamers with specificity for the food pathogens E. coli O157, Listeria monocytogenes, L. innocua, S. typhimurium and S. enteritidis. The aptamers against E. coli O157 and S. typhimurium were cloned and the sequences and the binding properties of these aptamers were analysed. The use of E. coli K12 as a target organism and the aptamer sequences presented in this study, have not previously been published in scientific literature. This is also the first report where the aptamers have been used in detection of live bacterial cells in yogurt.
    • Vitamin D and cardiometabolic disease risk: A RCT and cross-sectional study

      Mushtaq, Sohail; Hughes, Stephen F.; Agbalalah, Tari (University of Chester, 30/01/2017)
      Given the strong evidence for a beneficial role of vitamin D in diabetes and CVD pathogenesis, and the prevalence of vitamin D deficiency, vitamin D supplementation has been advocated for the prevention of cardiometabolic disease. To provide information on the effects of 5,000IU (125µg) vitamin D3 on cardiometabolic risk, a double blind, RCT in a cohort of overweight and obese UK adult males with plasma 25(OH)D concentration < 75nmol/L for a duration of 8 weeks was conducted. To the best of my knowledge, this is the first RCT to investigate the effect of 5,000IU (125µg) vitamin D3 on cardiometabolic markers in vitamin D insufficient, non-hypertensive and non-diabetic overweight and obese adult males.
    • Investigating the role of heatshock on diabetic wound healing

      Contractor, Taha (University of Chester, 2017-05)
      The increasing occurrence of diabetes in the general population as a result of over nutrition and increasingly inactive lifestyle has led to an obesity epidemic which is set to grow over time. With an ever increasing obese population type 2 diabetes and cardiovascular complications are set to become the major causes of human mortality. Chronic non healing wounds are a major cause of mortality and morbidity in patients with type 2 diabetes. They are predominantly caused by macrophage dysfunction and a lack of migration of fibroblasts into the wound. This study aimed to investigate diabetic wound healing through development of an artificial scratch assay. An in vitro scratch assay developed in WS1 cells. The effect of heat shock treatments from 39°C to 45° was tested to determine if cell migration increased; however, no significant difference was seen. Mitomycin C was used to determine if wound closure occurred as a result of cell proliferation and migration or migration alone. 10μg/ml of mitomycin C inhibited cell division by 79.9% without exhibiting cytotoxicity over a 12h period. The effect of hyperglycaemia and heat shock was also tested and showed no significant difference when compared to control conditions, suggesting that fibroblast migration in vivo is hindered through other factors such as debridement or macrophage dysfunction in the wound. GLUT4 is present in insulin sensitive organs (liver, adipose and muscle) and is the major glucose transporter responsible for the clearance of glucose from the blood after a meal, thus playing a central role in glucose homeostasis. Monocytes are precursors to macrophages and can easily be isolated from whole blood. They have also been shown to express GLUT4 in response to insulin and could be used as model to assess inflammation in diabetes. A glucose uptake assay was developed in U937 cells using a fluorescent glucose analogue, 2NBDG. 2NBDG fluorescence was shown to be competitively inhibited by increasing concentrations of glucose suggesting that 2NBDG enters the cell through glucose transporters. 2NBDG uptake was also assessed at different pH and in presence of membrane fluidizers (DMSO, benzyl alcohol and phenethyl alcohol). Extremes of pH significantly reduced cell viability and only at pH 4 was 2NBDG fluorescence significantly reduced. Treatment with DMSO showed that at high concentrations (≤ 1.56%) cell viability was reduced with a concurrent reduction in 2NBDG fluorescence. The effect of benzyl alcohol and phenethyl alcohol was foundto be insignificant at the concentrations and time points tested. The presence of GLUT4 was also determined by flow cytometry and Western blotting and found to be situated in the cytoplasmic region of the cell. This study indicates that monocytes and macrophages could be a potential therapeutic target to improve diabetic wound healing as they are a source of growth factors and cytokines that can bring about resolution of inflammation and it is their dysfunction in diabetic wounds that causes poor clinical outcomes.