• How men experience, understand, and describe masculinity: A phenomenological psychological analysis and photovoice exploration.

      Earnshaw, Deborah; University of Derby (University of Derby, 2018-04-17)
      This thesis is an examination of how men describe and experience masculinity. Psychological and sociological research has suggested that masculinity is malleable (Smiler, 2006), there are different versions or pluralities of masculinity (Connell 1995) and can be context-dependent (Gilbert & Gilbert, 2017). Often however hegemonic masculinity is considered the only type of masculinity, and is not flexible, especially when discussed on a social level (Cuthbert, 2015). Based on the researcher’s cultural, social and historical knowledge and understanding, masculinity is very different for people and so is understood, demonstrated and experienced in various ways. This research employed an unstructured interview design, incorporating photovoice, with five participants overall where each participant, except one, was interviewed twice. The first interview was researcher-led, with images provided by the researcher to be the focus for the participant. The second interview was participant-led, with the images provided by the participant to represent what they considered to be masculine or represented masculinity in their everyday life. The data collected was analysed using a combination of phenomenological methods; Descriptive Phenomenological Psychology (Giorgi, 2009) and Hermeneutic Phenomenology (van Manen, 2016). The thesis is presented in two halves. The first is researcher-led and draws on hermeneutic psychology and presents three themes were found from the first interviews: Hegemonic Masculinity with Traditional Masculinity, Characteristics and Non-Conformity; Societal Influence with Culture, Image and Media; and Feminism and Women. The second part of the thesis is participant-led, and draws out the descriptive phenomenological aspects by presenting each individual’s interaction with their chosen images and their ensuing descriptions of masculinity illustrated by them. Themes in this context are individually related rather than demonstrated through a cross-case analysis. Findings demonstrated masculinity as an individual identity, with a social expectation of how men should behave and portray themselves. The way it is perceived, understood, experienced and described is different for each person, as was demonstrated here with the participants’ second interviews. Future research should consider expanding research to include more on everyday factors, such as the use and influence of social media, the projection of masculinity throughout a man’s life, and how men and women both aid in the creation and maintenance of masculinity.
    • The impact of Polycystic Ovary Syndrome (PCOS) on quality of life: exploration, measurement and intervention.

      Williams, Sophie; University of Derby (2016)
      Polycystic Ovary Syndrome (PCOS) is one of the most common endocrine disorders amongst women, estimated to affect one out of 10 women. Symptoms include infertility, obesity, alopecia, acne, hirsutism and menstrual irregularities. Women with the syndrome are also more likely to experience co-morbid physical and psychological conditions such as diabetes, heart disease, endometrial cancer and also depression and anxiety. PCOS has also been found to have a negative impact on quality of life. This thesis aimed to further understanding, and improve quality of life of women with PCOS in the UK. To achieve this, the thesis aimed to investigate and identify how women with PCOS in the UK perceive and define their quality of life and to further understanding of the day-to-day experience of living with PCOS. Moreover, in order to measure quality of life, it aimed to develop and validate a UK disease-specific quality of life measure for women with PCOS. It also aimed to identify, develop and test a pilot intervention to increase quality of life in women with PCOS. To achieve these aims a mixed-methods approach was taken employing a variety of data generation and collection methods including: photovoice, online Skype™ interviews; LimeSurvey and Qualtrics. The findings of this thesis emphasise that PCOS has a negative impact on quality of life; encompassing psychological, social, environmental, and physical domains of quality of life. Women with PCOS who experienced the symptoms of infertility, hirsutism, weight, alopecia, skin discolouration, skin tags and mood swings had significantly lower scores of overall quality of life than those women who did not experience the symptoms. In addition, those women with PCOS who had a diagnosis of anxiety and/or depression had reduced quality of life. The dissemination of these findings will enable health care professionals to better understand the experience of living with PCOS and its impact on quality of life. Moreover, this thesis identifies many areas for future research which will enable a better understanding of the impact of PCOS on quality of life. Finally, this thesis makes recommendations for clinical practice which include improvement of support from health care professionals for women with PCOS in order to help them better manage their symptoms, and therefore improve their overall quality of life.
    • The influence of caffeine expectancies on simulated soccer performance and perceptual states

      Hooton, Andy; Sheffield, David; Higgins, Matthew; Shabir, Akbar (University of DerbyThe University of Derby, School of Human Sciences, 2020-10)
      Caffeine (CAF) is the most widely consumed ergogenic substance in sport and has been reported to improve various attributes associated with successful soccer performance including, endurance capacity, gross motor skill performance and cognitions. These benefits are typically ascribed to pharmacological mechanisms (i.e. central nervous and peripheral tissue stimulation). However, the psychological and perceptual permutations that are associated with CAF expectancies are largely unaddressed in most experimental designs but could be as important as CAF pharmacology in understanding if/how CAF elicits an ergogenic response on sport performance. As the consumption of pharmacologically active CAF may not be necessary in observing a CAF associated ergogenic response, this body of work may prove beneficial to individuals suffering from pre-existing health concerns (e.g. hypertension, genetic polymorphisms, depression, insomnia etc.), CAF habituation, and those participating in late evening sports competitions whereby CAF consumption may impair sleep quality/duration. The main aim of this thesis is to evaluate and explore the psychobiological effects of expectancies associated with oral CAF consumption on various facets of simulated soccer performance and perceptual states. This was achieved via completion of the following objectives: (1) conducting a systematic review and meta-analyses pertaining to literature exploring the potential psychobiological effects of CAF on sport and exercise (A) and cognitive performance (B) (2) exploration of the potential psychobiological effects of CAF on high-intensity intermittent endurance capacity, reaction time and soccer skill proficiency (3) exploration of the potential psychobiological effects of CAF on perceptual fatigue and mood states (4) exploration of the potential psychobiological effects of CAF on subjective perceptions using the double-dissociation design but in particular the mixed methods approach (Brooks et al., 2015). There remains a severe under representation of the mixed methods design in the literature pertaining to the phenomenon of CAF expectancies on sport, exercise, and cognitive performance. The mixed methods design and associated triangulation of quantitative and qualitative data is fundamental to validly compare CAF’s psychological versus pharmacological impetus. Prior to this thesis, only two other studies (Beedie et al., 2006; Duncan et al., 2009) implemented the mixed methods design, with neither providing a rigorous account of methodological decisions, researcher reflexivity, and/or evidence of applying an epistemological framework. These factors were incorporated within the current thesis and improved the quality of data collection, analysis, and reflection. The results of our, novel, meta-analyses demonstrated that all studies exploring the psychobiological effects of CAF on sport and exercise performance displayed a beneficial effect (p=0.01) with an overall moderate effect size (Cohen’s d (ES): 0.40) observed. In contrast, no significant effect was observed for studies exploring the psychobiological effects of CAF on cognitive performance (p=0.142) with a small effect size (ES=0.1) observed. Though, due to significant methodological heterogeneity associated with studies exploring the psychobiological effects of CAF on cognitive performance, any associated implications here should be taken with caution. Experimental study 1 explored the influence of CAF expectancies on facets of simulated soccer performance (e.g. exercise capacity, reaction time and passing ability (LSPT)) and perceptual states via utilisation of a mixed-methods approach and double-dissociation design. Exercise capacity was greater (p<0.05) for CAF psychology (given placebo (PLA)/told CAF) (623 ± 117 s) versus pharmacology (given CAF/told PLA) (578 ± 99 s) with all participants running longer during psychology. This benefit appeared to be driven by CAF expectancies and reductions in perceptual effort. Interestingly, positive perceptions for told CAF conditions appeared to impair BATAK performance via potential CAF over reliance. In contrast, negative perceptions possibly facilitated BATAK performance via augmented conscious effort. A similar trend to BATAK was observed for LSPT performance. Following the completion of experimental study 1 it became apparent that the techniques used to modulate expectancies across experimental conditions (i.e. told PLA/CAF groups) here or any other study with a primary aim of exploring the influence of CAF expectancies on sport, exercise and/or cognitive performance, require validation. This was the premise of experimental study 2. No meaningful findings were observed from baseline to post-intervention across any outcome measure during experimental study 2. This lack of effect may be related to environmental factors, whereby individuals completed trials in classrooms and/or a home cinema, prior to lectures/seminars and/or social interactions, respectively. In contrast, participants in experimental study 1 were administered the appropriate expectancy modulating techniques after they had perceived to consume PLA or CAF within an environment that necessitated an immediate importance for CAF (e.g. prior to exercise performance). In summary, the novelty and original contribution of the current body of research entails: completion of a systematic review and meta-analyses pertaining to the influence of CAF expectancies on sport, exercise and cognitive performance; assessment of the influence CAF expectancies may have on simulated soccer performance; the implementation of a mixed methods approach and double dissociation design; an in depth rationale, description and set of instructions for the utilisation of the mixed methods approach in any future research, including the use of an epistemological framework; a summary of ecological factors that are fundamental in understanding the phenomenon of CAF expectancies across sport and exercise performance. With respect of the main findings from the experimental data contained in this thesis: the benefits associated with CAF expectancies may better suit tasks that entail lesser cognitive/skill specific attributes but greater gross motor function (e.g. cycling, weightlifting, running etc.) and this is likely due to reduced perceptual effort. Moreover, future studies aiming to validate expectancy modulating techniques or generally assessing expectancies should provide a greater immediate importance for CAF and this may be achieved by replicating environmental and/or psychosocial conditions associated with sport performance (e.g. the utilisation of a performance measure) and the perception for CAF consumption.
    • Late quaternary fluvial system response to climatic change over the past 200ka on Mallorca, Illes Balears.

      Thompson, Warren; University of Derby (University of Derby, 2017-11-02)
      Outcrops of late Quaternary deposits along the north–east coast of Mallorca were examined, and a high resolution chronological framework established using optically stimulated luminescence of quartz and feldspar grains. Coastal sections at Es Barrancar and Cala Mata record a complex succession of alluvial fan deposition with a minor aeolian component, mainly deposited during the last two glacial cycles. For the last ~200ka different orbital configurations appear to have produced a series of subtly different climatic scenarios that resulted in great variations in the regional availability of moisture. In turn, each of these climatic scenarios set in motion a distinct set of sedimentary processes, which imprinted themselves upon the character of fluvial system response recorded in the alluvial archives on Mallorca. Within the resulting complex of sediments were units of fluvially reworked aeolianite which, although truncated in places, were traceable laterally along wide sections of the sea front outcrops of both fan systems. These archives yielded 47 new OSL and pIRIR290 ages which suggest a number of previously unrecognised periods of enhanced fluvial activity. Luminescence ages identify significant ephemeral fluvial activity taking place during MIS 6, MIS 5 sub-stages d/c, b/a, and across the MIS 5/4 boundary. Another major phase of reworking corresponds with the middle part of MIS 3, and continued sporadically into the Last Glacial Maximum. These fluvial reworking events have been interpreted as having taken place during cold arid climatic intervals, when vegetation was at a minimum, precipitation was low and displayed a much higher degree of seasonality, enhancing the effects of runoff.
    • Limitations and trainability of the respiratory system during exercise with thoracic loads

      Faghy, Mark; University of Derby (2016-02-01)
      Thoracic loads (i.e., a heavy backpack) commonly used in occupational and recreational settings significantly challenge human physiological systems and increase the work of breathing, which may promote respiratory muscle fatigue and negatively impacts whole body performance during physical tasks. Accordingly this thesis: (Chapter number: II) designed a laboratory based protocol that closely reflects occupational demands and (III) assessed the effect that load carriage (LC) has upon physiological and respiratory muscle function. Consequently the thesis addressed (IV) acute, (V) chronic and (VI) functional inspiratory muscle loading strategies to assess the limitations and trainability of the respiratory muscles to load carriage performance. The novel laboratory protocol, performed wearing a 25 kg backpack load, combined submaximal load carriage (LC; 60 min treadmill march at 6.5 km·h-1) and self-paced time trial exercise (LCTT; 2.4 km) to better reflect the physiological demands of occupational performance (between trials mean difference -0.34 ± 0.89 min, coefficient of variation 10.5%). Following LC, maximal inspiratory muscle pressure (PImax) and maximal expiratory muscle pressure (P¬Emax) were reduced by 11% and 13% respectively (P<0.05), and further by 5% and 6%, respectively (P< 0.05), after LCTT. Acute inspiratory loading (2 × 30 forced inspiratory efforts 40% PImax) following an active warm-up (10 min lactate turnpoint) failed to improve LCTT despite a transient increase in PImax of ~7% (P<0.05). Chronic inspiratory loading (6 wk, 50% PImax, 30 breaths twice daily) increased PImax (31%, p<0.05) reduced HR and perceptual responses post-LC, and improved LCTT (8%, P< 0.05) with no change in a placebo control. Combining IMT with functional core muscle exercises improved PImax and LCTT by 7% and 4% respectively (P< 0.05), which was greater than traditional IMT alone. Acute, chronic and functional inspiratory muscle loading strategies did not protect against respiratory muscle or locomotor muscle fatigue during LC and LCTT.
    • Oral Human Papillomavirus (HPV) Prevalence and Abundance in the UK Young Adult Population

      Marsh, Elizabeth; Knight, Gillian; Whitton, Aimee (University of DerbySchool of Biomedical & Forensic Science, 2021)
      BACKGROUND: The incidence rates of HPV positive oropharyngeal cancers (OPSCCs) are on the rise, yet oral HPV prevalence rates in clinically healthy populations are poorly understood. To determine the risk of healthy adults developing OPSCCs, first we must establish oral HPV prevalence, viral load, persistence, and clearance rates in healthy populations to understand the link between oral HPV and OPSCCs. This is even more pertinent within the young adult population as the HPV vaccination programme has been shown to reduce cervical cancer incidence but not OPSCC incidence. Therefore, other factors that could affect oral HPV contraction and OPSCC development need investigation such as population demographics, lifestyle risk behaviours, HPV screening methods and vaccination status. OBJECTIVES: The aims of this study were centred on establishing reproducible and sensitive HPV screening methods for the detection of oral HPV in clinically healthy young adults, for determining prevalence and abundance, and establishing if oral HPV was influenced by vaccination status, demographics, and lifestyle risk behaviours. METHODS: The study established a novel and sensitive real-time PCR HPV consensus screening method for the detection of multiple HPV subtypes in the oral cavity of 408 clinically healthy UK-based young adults (92.01%; 18-25 years old in 2016-17). HPV positive or undetermined samples were then screened using qPCR for HPV subtypes, HPV-6, HPV-11, HPV-16, HPV-18. All results were analysed alongside vaccination status, demographics and lifestyle risk behaviour data collected via questionnaire. RESULTS: An oral HPV prevalence rate of 22.79% was found, with HPV-16 being the most prevalent and abundant subtype at 19.12%; 1.08x105 copies/million cells, followed by HPV-18 at 1.72%; 1.89x104 copies/million cells, HPV-6 at 0.49%; 4.50x102 copies/million cells and HPV-11 at 0.25%; 1.06x102 copies/million cells. Unknown HPV subtypes were detected in 2.21% of the cohort. Oral HPV was found to be significantly associated with open-mouth kissing (p <.001), oral sex (p = .049), masturbation in males (p = .020), sexual intercourse (p = .026), sexual activity diversity (p = .043), frequent smoking (p = .024), wine drinking (p = .045) and drinking ≥2 types of alcohol per sitting (p = .015), especially in males (p = .023). HPV-16 was significantly associated with masturbation (p = .004), whilst there was a reduction in viral load in vaccinated individuals, but this was not statistically significant. CONCLUSIONS: Oral HPV is prevalent in the young adult UK population, especially HR-HPV subtype HPV-16, questioning the efficacy of the HPV vaccination on reducing oral prevalence. However, HPV vaccination may instead influence oral HPV viral load, but further research is required, demonstrating the importance of measuring both presence and abundance. Oral HPV prevalence did appear to be influenced by sexual practice, including open-mouth kissing and oral sex, but less so by smoking and alcohol consumption, reaffirming the link between oral HPV and OPSCCs.
    • Pain responses in athletes:

      Thornton, Claire; University of Derby (2018-09-04)
    • Problematic painkiller use in the general population: a multi-national comparison exploring the role of accessibility of painkillers and psychological factors

      Said, Omimah; University of Derby (2018)
      Problematic painkiller use is a large and increasing problem worldwide, leading to serious physical, psychological and social consequences. Existing research indicates that accessibility of painkillers and psychological factors could have a role in problematic painkiller use. To clarify, accessibility refers to ease of obtaining painkillers, whilst psychological factors refer to individual-level processes and meanings that influence mental states (Upton, 2013). However, there have been few studies conducted, and these studies have focused mainly on either clinical samples or women with childbirth pain. Hence, the role among the general population is less clear. The aim of the present thesis was therefore to focus on the role of accessibility and psychological factors in problematic painkiller use among the general population. Three studies were conducted: one study compared the general population of the UK (N = 295) and Egypt (N = 420) regarding the role of accessibility of painkillers and psychological factors, including attitudes and beliefs towards pain, painkillers, self-medication and alternative methods of pain relief; another study was a multi-national comparison of these variables among the general population of more countries including Germany (N = 217), USA (N = 146), Australia (N = 93) and China (N = 76); another study focused on the role of psychological factors over time among the UK general population (N = 529), specifically attitudes and beliefs towards painkillers, as well as pain catastrophising, pain acceptance, pain self-efficacy and alexithymia. In these studies, the role of accessibility and psychological factors was investigated using online surveys, with participants aged 18 years or over, who experienced pain in the last month, used over-the-counter (OTC) or prescription painkillers in the last month, and were residents of the countries concerned. An additional study was conducted to develop 14-item versions of the Survey of Pain Attitudes-Brief (SOPA-B-14) and the Pain Medication Attitudes Questionnaire (PMAQ-14), which also tested the validity of these scales. Results Accessibility of painkillers and psychological factors predicted problematic painkiller use. However, there were several differences between the countries regarding the particular role of these factors. In the longitudinal study of the UK general population, changes in psychological factors were found over time, but attitudes and beliefs about withdrawal from painkillers was the only psychological factor that predicted problematic painkiller use over time. In addition, testing the validity of the SOPA-B-14 and PMAQ-14 showed that these scales were valid. The present research provided understanding regarding the role of accessibility and psychological factors in problematic painkiller use among the general population, and the role of psychological factors over time. Based on this understanding, interventions focusing on accessibility and psychological factors should be developed to reduce problematic painkiller use, but tailored to the particular factors that were predictors for each country. The present research also developed a valid SOPA-B-14 and PMAQ-14, therefore these scales can be used rather than the full versions to make assessment easier.
    • Psychophysiological and emotional antecedents of climbing performance

      Giles, David; University of Derby (University of Derby, 2017-09-12)
      Recreational sport climbing is characterised by self-selected route choices, which place participants under both physiological and psychological stress. This thesis is comprised of four studies, each conducted with experienced climbers, exploring subjective psychological, objective psychophysiological and behavioural responses to anxiety-inducing stressors. Studies One and Two explored the means of protecting a climber in the event of a fall and the relative difficulty of a route. Significant and meaningful differences in self-reported anxiety and climbing performance were found in both studies. However, notably, psychophysiological measures of anticipatory heart rate and cortisol did not result in meaningful differences. Results suggested that situations, atypical of participants’ normal recreation sessions, with an increased likelihood of a climber falling or being unable to complete the route, were likely to be evaluated as threatening, elicit a negative emotional response and disrupt performance. However, the quantitative methods employed in Studies One and Two did not provide an explanation of the processes underlying participant’s anxious response and disrupted performance. Consequently, Study Three qualitatively explored individual experiences of climbers, with a focus on psychological factors that influence performance. The defining characteristics of lead climbing were discussed, as were the potential for taking falls, and/or the anticipation of falling. Further, interviewees described the choices they make, in order to increase or decrease the physical, psychological and technical challenges present. Critically, the choices made by a climber appear to potentiate or limit opportunities to perform optimally. Climber’s decisions were mediated by a number of antecedents, including a climber’s background in the sport, climbing partners and training status. Data suggests that while decisions made by the climbers allow them to engage with the sport on their own terms and exert a level of control over the challenges of their climbing sessions, it is often at the expense of performance. Interestingly, while interviewees were aware of techniques to reduce anxiety and improve performance, few regularly used these in training. Study Four examined the effectiveness of clip drops and repeat practice to reduce anxiety. Results indicated that neither technique resulted in reduced anxiety or improved performance when compared to the control group. While there were small differences in the success rate of participants in the intervention groups, they were less anxious and interpreted their level of self-confidence as more positive, compared to control, it was not possible to differentiate between the two interventions. However, when the combined means were considered there were significant and meaningful differences observed in the post-intervention red-point ascent compared to the initial on-sight. This thesis highlights the difficulty that arises in attempting to quantitatively examine anxiety. While there might not be (a) anxiety in climbers or (b) quantifiable differences between climbers of different abilities, it may be that what is possibly ‘noise’ in data arises due to weaknesses in the markers themselves. The findings of Study Three provide evidence of the true nature of anxiety for climbers, which was not evident from the quantitative markers; as well as the lengths climbers will go to, to avoid anxiety. Climbers’ responses to anxiety were individualised, consequently, generalised interventions may have a limited effect on reducing anxiety to a level which supports performance improvements. It may be that an individualised approach to anxiety reduction and avoidance behaviours has a more significant impact on performance improvement than any of the latest training programmes, equipment or nutritional strategies.
    • Risk-taking and expenditure in digital roulette: Examining the impact of tailored dynamic information and warnings on gambling attitudes and behaviours.

      McGivern, Paul R.; University of Derby (2018-07-20)
      Digital gambling is the fastest growing form of gambling in the world (Reilly & Smith, 2013a). Technological advancements continually increase access to gambling, which has led to increased social acceptance and uptake (Dragicevic & Tsogas, 2014) with Roulette being among the most popular games played both online and on Electronic Gaming Machines. In response, gambling stakeholders have drawn on the structural characteristics of gambling platforms to develop and improve Responsible Gambling (RG) devices for casual gamblers. Many RG data-tracking systems employ intuitive ‘traffic-light’ metaphors that enable gamblers to monitor their gambling (e.g. Wood & Griffiths, 2008), though uptake of voluntary RG devices is low (Schellinck & Schrans, 2011), leading to calls for mandatory RG systems. Another area that has received considerable RG research focus involves the use of pop-up messages (Auer & Griffiths, 2014). Studies have examined various message content, such as correcting erroneous beliefs, encouraging self-appraisal, gambling cessation, and the provision of personalised feedback. To date, findings have been inconsistent but promising. A shift towards the use of personalised information has become the preferred RG strategy, though message content and timing/frequency requires improvement (Griffiths, 2014). Moreover, warning messages are unable to provide continuous feedback to gamblers. In response to this, and calls for a ‘risk meter’ to improve monitoring of gambling behaviours (Wiebe & Philander, 2013), this thesis tested the impact of a risk meter alongside improved pop-up warning messages as RG devices for within-session roulette gambling. The thesis aimed to establish the optimal application of these devices for facilitating safer gambling behaviours. In support of the aims of RG research to evaluate the impact of devices on gambling attitudes and behaviours, the Elaboration Likelihood Model was identified as a suitable framework to test the proposed RG devices (Petty & Cacioppo, 1986). Both the interactive risk meter and pop-up messages were developed based on existing methods and recommendations in the RG literature, and examined via a series of laboratory-based roulette simulation experiments. Overall, results found the risk meter to be most effective when used as an interactive probability meter. Self-appraisal/Informative pop-up warnings were examined alongside expenditure-specific and hyrbid warnings. Findings showed that hybrid messages containing both types of information to be most effective, with optimal display points at 75%, 50%, 25% and 10% of remaining gambling credit. The final study tested both optimised devices (probability meter and hybrid messages). Results showed that using both RG devices in combination was most effective in facilitating reduced gambling risk and early within-session gambling cessation. Findings support the use of personalised, interactive RG devices using accurate context-specific information for the facilitation of safer gambling. The ELM was shown to be an effective model for testing RG devices, though findings suggested only temporary shifts in attitude change and a lack of impact on future gambling intentions. Overall, support for the implementation of RG devices that facilitate positive, temporary behaviour change that do not negatively impact on broader gambling attitudes or gambling enjoyment. Implications for theory, implementation, and RG frameworks are discussed, alongside recommendations for future research.
    • Stability of protein-based drugs: Herceptin a case study

      Shropshire, Ian Michael; University of Derby (2016)
      There is a lack of stability data for in-use parenteral drugs. Manufacturers state a shelf-life of 24 hours for infusions based on microbiological contamination. The lack of data is of particular significance with protein-based drugs where action is determined by their complex structure. A range of techniques are required to assess stability, including biological assessment to support other data. There has been an increase in published data but often the few studies that address in-use stability are incomplete as they do not employ biological assessment to assess potency. Trastuzumab is an antibody-based drug used to treat cancers where the Epidermal Growth Factor Receptor 2 (HER2) is over expressed or over abundant on the cell surface. Trastuzumab infusions have been assigned by the manufacturer to be stable for 24 hours at temperatures not exceeding 30 oC. If stability is shown beyond this point it would enable extended storage and administration. To this end, methods were selected and developed with biological assessment central to the approach to assess clinically relevant infusion concentrations (0.5 mg/mL and 6.0 mg/mL) and a sub-clinical infusion concentration (0.1 mg/mL). This may enhance instability and provide opportunity to study degradation. A Cell Counting Kit CCK8 (Sigma Aldrich) was ultimately adopted as a basis for a colorimetric assay to assess cell viability. Attenuated Total Reflectance Infra-Red Spectroscopy and Size Exclusion Chromatography methods were developed to evaluate secondary structure and aggregation respectively. These methods were applied to a shelf-life study (43 days) as a collaboration with Quality Control North West (NHS) and Clatterbridge Centre for Oncology NHS Foundation Trust, Clatterbridge Hospital. There was no evidence of degradation and no loss efficacy for clinically relevant infusions (0.5 mg/mL and 6.0 mg/mL) over 43 days, whilst the sub-clinical infusions (0.1 mg/mL) developed particles after 7 days of storage between 2 oC and 8 oC. Furthermore, evidence of stability at day 119 gave increased confidence for the data from earlier time points. This work assisted in the shelf-life being recommended to be extended to 28 days for Trastuzumab stored in polyolefin IV bags at concentrations between 0.5 mg/mL and 6.0 mg/mL with 0.9% saline between 2 oC and 8 oC. However, infusions with concentrations below 0.5 mg/mL were not recommended for storage.
    • Transforming a Research Concept into Commercial Practice: Addressing the ‘Hurdles’ of Single-Species eDNA-based Detection

      Sweet, Michael; Robinson, Louise; Burian, Alfred; Troth, Christopher (University of Derby, 2020-01-15)
      The use of environmental DNA (eDNA) for measuring and monitoring biodiversity has been identified as a novel molecular based method to complement more commonly utilised traditional ecological sampling techniques. It is a time and cost-efficient technique, which is rapidly advancing due to the capabilities of low eDNA detection levels. As the efficiency of the technique has increased, commercial organisations and end-users have gained a greater interest in its application. Despite this, the technique is currently only commercially available from a select few service providers. In the UK, the main target species for commercial scale eDNA-based detection is the great crested newt (Triturus cristatus). Interest has now been sparked for the development of eDNA assays to detect various other species, both for use as a regulated informative tool and a conservation aid. However, many recent studies have highlighted various limitations associated with the use of eDNA-based detection and this appears to be hampering commercialisation of this tool. eDNA-based detection methods remain relatively underdeveloped and un-validated for use as reliable and accurate widespread monitoring programs and other such applications. Here, the so called ‘hurdles’ associated with the development and validation of eDNA-based methods and its use as a fully available commercial service are reviewed and addressed, in order to develop and validate a commercially applicable eDNA assay for the endangered white-clawed crayfish, Austropotamobius pallipes, as a target organism. When designing novel species-specific assays, detailed validation steps need to be undertaken, ensuring they perform under various conditions, habitats, and which sampling methods should be utilised. Currently, more traditional methods used to asses populations of white-clawed crayfish (such as trapping and hand searching) are becoming increasingly more difficult to undertake as the species become rarer and populations more fragmented. Such techniques are therefore expensive (with regard to time spent surveying) and often result in low probability of detection. A new species-specific qPCR assay to detect white-clawed crayfish was developed and tested under various conditions both ex-situ (laboratory and mesocosms) and in-situ (ponds and rivers) to explore the optimum sampling strategy giving the most reliable results. Experiments were also conducted on a wider scale to determine the impact of DNA degradation and seasonal influence on eDNA persistence. Interestingly, this thesis illustrates that sample collection choice is not simple, and the ‘best’ methodology was shown to vary between habitat type. This indicates that great care should be taken when designing any such assays and implementing them in the field. Furthermore, this study highlights that a ‘standard operating procedure’ for eDNA-based detection in the commercial sector may not be possible and this will have to be explored on an assay by assay basis. Alongside case studies from real-world application of the technique, recommendations are made on how this novel eDNA assay can be used for the commercial practice of white-clawed crayfish assessment.