• Activating KIR Haplotype Influences Clinical Outcome Following HLA-Matched Sibling Hematopoietic Stem Cell Transplantation.

      Heatley, Susan L.; Mullighan, Charles G.; Doherty, Kathleen; O'Connor, Geraldine M.; Hahn, Uwe; Szer, Jeff; Schwarer, Anthony; Bradstock, Kenneth; Sullivan, Lucy C.; Bardy, Peter G.; et al. (Wiley, 2018-06-25)
      Natural killer cells are thought to influence the outcome of hematopoietic stem cell transplant (HSCT), impacting on relapse, overall survival, graft versus host disease and the control of infection, in part through the complex interplay between the large and genetically diverse killer immunoglobulin-like receptor (KIR) family and their ligands. This study examined the relationship between KIR gene content and clinical outcomes including the control of opportunistic infections such as cytomegalovirus in the setting of human leucocyte antigen (HLA)-matched sibling HSCT in an Australian cohort. The presence of the KIR B haplotype which contain more activating receptors in the donor, in particular centromeric B haplotype genes (Cen-B), was associated with improved overall survival of patients with acute myeloid leukemia (AML) undergoing sibling HSCT and receiving myeloablative conditioning. Donor Cen-B haplotype was also associated with reduced acute graft versus host disease grades II-IV whereas donor telomeric-B haplotype was associated with decreased incidence of CMV reactivation. In contrast, we were not able to demonstrate a reduced rate of relapse when the donor had KIR Cen-B, however relapse with a donor Cen-A haplotype was a competing risk factor to poor overall survival. Here we show that the presence of donor activating KIR led to improved outcome for the patient, potentially through reduced relapse rates and decreased incidence of acute GvHD translating to improved overall survival.
    • Application of immunological methods for the detection of species adulteration in dairy products

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

      Schoeler, Natasha; Simpson, Zoe; Whiteley, Victoria; Nguyen, Patty; Meskell, Rachel; Lightfoot, Kathryn; Martin-McGill, Kirsty; Olpin, Simon; Ivison, Fiona
      Objective: Biochemical assessment is recommended for patients prior to initiating and following a ketogenic diet (KD). There is no published literature regarding current practice in the UK and Ireland. We aimed to explore practice in comparison to international guidelines, determine approximate costs of biochemical testing in KD patients across the UK and Ireland, and promote greater consistency in KD services nationally. Methods: A survey was designed to determine the biochemical tests requested for patients at baseline, 3-, 6-, 12-, 18- and 24-months+ on KD. The survey was circulated to 39 centres across the UK and Ireland. Results: 16 centres completed the survey. Full blood count, electrolytes, calcium, liver function tests (LFTs), lipid profile and vitamin D were requested at all centres at baseline, in keeping with international guidelines. Bicarbonate, total protein and urinalysis were less consistently requested. Magnesium and zinc were requested by all centres, despite not being specifically recommended for pre-diet evaluation in guidelines. Urea and electrolyte profiles and some LFTs were consistently requested at follow-up, in accordance with guidelines. Other LFTs and renal tests, full blood count, lipid profile, acylcarnitine profile, selenium, vitamin D and urinalysis were less consistently requested at follow-up. The mean costs of the lowest and highest number of tests requested at baseline in our participating centres was £167.54 and £501.93; the mean costs of the lowest and highest number of tests requested at 3-month follow-up was £19.17 and £450.06. Significance: Biochemical monitoring of KD patients varies widely across the UK and Ireland and does not fully correspond to international best practice guidelines. With an ongoing drive for cost-effectiveness within healthcare, further work is needed to streamline practice whilst ensuring patient safety.
    • Chapter Ten: Handling and Restraint of Small Ruminants

      McLennan, Krista M.; Chapman, Stella; University of Chester; University Centre Hartpury (Wiley, 2017-11-03)
      Sheep (Ovis aries) were one of the first mammals to be domesticated by humans; however the exact timeline of events has been unclear. The use of mitochondrial DNA testing has recently made it possible to trace back the ancestry of many animals including cattle, horses, pigs and goats and evidence suggests that the number of wild progenitors for these species is limited; however, with the sheep this is not the case and it is thought that a large number of wild ancestral species and subspecies exist (Hiendleder et al. 2002). Archaeological findings have traced the sheep back to 11000 and 9000 BC in Mesopotamia, with the most common hypothesis being that Ovis aries descended from the Asiatic (Ovis orientalis) species of mouflon. Many studies have looked at the ancestry of sheep and there has been conflicting evidence with regards to the numbers of ancestors. It is now thought that three major groups of Eurasian wild sheep (mouflon, urial and argali) are the ancestors of the domestic sheep and it is these groups that are believed to have contributed to specific breeds (Hiendleder et al. 2002).
    • Chapter Twelve: Handling and Restraint of South American Camelids

      McLennan, Krista M.; Chapman, Stella; University of Chester; University Centre Hartpury (Wiley, 2018-01-01)
      Members of the camelid family evolved to live in arid and mountainous areas. This chapter will focus on what are known as the New World species of camelid, whose habitat mainly covers the Andes regions of South America. Four camelids can be found in South America, namely: Guanacos (Lama guanicoe), vicunas (Lama vicugna), llamas (Lama guanicoe glama) and alpacas (Vicugna pacos). The two wild forms, the guanaco and the vicuna diverged from a common ancestor approximately two million years ago; an event unrelated to domestication. Due to hybridisation the exact process of domestication has been controversial; however, recent genetic analysis has suggested that the alpaca is the domesticated form of the vicuna and the llama is the domesticated form of the guanaco (Kadwell et al. 2001). Domestication is thought to have taken place some 6000 years ago (Wheeler, 1995) when a predominant herding economy based on llama and alpaca was established at Telarmachay (a region of the Peruvian Andes). Archaeological evidence suggests that both llamas and alpacas were part of a sacrificial rite in South American culture and were key to the expansion of the Inca Empire some 500 years ago (Bonacic, 2011). Physically (apart from size) there is little difference between the llama and alpaca, which is a result of deliberate hybridisation between the two species over the past 35 years. Whilst the alpaca and llama still play an important role in their countries of origin, they are also viewed worldwide as: pets, exotic animals, livestock, zoo animals and wild animals.
    • Clobazam add-on therapy for drug-resistant epilepsy

      Bresnahan, Rebecca; Williamson, John; Martin-McGill, Kirsty J.; Michael, Benedict D.; Marson, Anthony G. (Wiley, 2019-10-22)
    • Cognitive function and disability in late Life: An ecological validation of the 10/66 battery of cognitive tests among community dwelling older adults in south India

      Krishna, Murali; Beulah, Eunice; Jones, Steven; Sundaracharj, Rajesh; Saroja, A.; Kumaran, Kalyanaraman; Karat, Samuel C.; Prince, Martin; Fall, Caroline H. D.; University of Chester (Wiley, 2015-12-17)
      Key Points • 10/66 cognitive tests are well suited for identification of older adults with cognitive and functional impairment at a population level in LMIC setting. • Lower scores on individual domains of the 10/66 battery of cognitive tests are associated with higher levels of disability and functional impairment. • It is feasible to administer 10/66 cognitive assessments in participant's own homes in India. • 10/66 cognitive tests are education and culture fair, suitable for use in population based research in India.
    • Dietary approaches for patients with heart failure and diabetes

      Butler, Thomas; Georgousopoulou, Ekavi N; Mellor, Duane (Wiley, 2018-08-20)
    • Distribution, status and recent population dynamics of Alpine ibex Capra ibex in Europe

      Brambillla, Alice; von Hardenberg, Achaz; Nelli, Luca; Bassano, Bruno; University of Zurich, University of Chester, University of Glasgow, Gran Paradiso National PArk (Wiley, 2020-04-20)
      1. Despite its recent successful and well-documented reintroduction history, a comprehensive and current update of the distribution and status of the Alpine ibex Capra ibex is lacking. As some concerns persist about its conservation, a status update appears essential for future conservation and management strategies on a large scale. 2. We provide an exhaustive update of the geographic range of the species, alongside estimates of its current abundance and population trends from 2004 to 2015. 3. We gathered census and distribution data for all the Alpine ibex colonies from management authorities and research groups that monitor them in different countries, and from the literature and publicly available reports. We produced a distribution map, reported the number of individuals observed in the most recent censuses, and estimated global, national, and local population trends using Bayesian hierarchical models. 4. Our model estimated that there were a total of 55297 Alpine ibex in the Alps in 2015 (lower 95% Credible Interval [CrI]: 51157; upper 95% CrI: 62710). The total number of individuals appears to have increased slightly over the last 10 years from the 47000-51000 estimated in previous reports. Positive population trends were observed in Switzerland and Italy, while no trend was apparent in France. For Austria, Germany, and Slovenia, there were insufficient data to estimate a trend. The slopes of the colonies’ trends were positively correlated with the year of colony foundation. 5. The geographic range of the Alpine ibex does not seem to have increased in size in recent years, although the accuracy of the spatial data varies among countries. 6. The periodic and standardised collection of census data for all colonies and a common policy of data-sharing at a European level appear essential for monitoring the global trend of this species and for planning balanced conservation and management actions.
    • Female clustering in cockroach aggregations – a case of social niche construction?

      Stanley, Christina R.; Preziosi, Richard F.; Liddiard Williams, H.; University of Chester, University of Manchester, Manchester Metropolitan University (Wiley, 2018-07-18)
      Individuals in groups can suffer costs through interactions with adversarial or unknown conspecifics. Social niche construction allows individuals to buffer such potential costs by only engaging in preferred associations. This may be particularly beneficial in insect aggregations, which are often large and highly fluid. However, little is known regarding the structuring of such aggregations. Here we use social network analyses to test for fine-scale social structure in resting aggregations of the sub-social cockroach Diploptera punctata and to explore the social pressures that contribute towards such structure. We showed that females were significantly more gregarious than males and formed the core of the proximity network, thus demonstrating a higher level of social integration. This fine-scale structure is likely to result from females displacing males; females initiated most displacements whilst males received the majority. We explain this behaviour in terms of social niche construction by showing that females received significantly fewer approaches and investigations at more female-biased local sex ratios. We therefore suggest that female social clustering occurs in this, and presumably other, species to reduce potential costs associated with male harassment. This demonstrates how social niche construction can lead to higher level social structure; we suggest this approach could be used across a range of species in order to improve our understanding of the evolution of sociality.
    • Heat shock proteins form part of a danger signal cascade in response to lipopolysaccharide and GroEL

      Davies, Emma L.; Bacelar, Maria M. F. V. G.; Marshall, Michael J.; Johnson, E.; Wardle, T. D.; Andrew, Sarah M.; Williams, John H. H.; University of Chester ; University of Chester ; Charles Salt Centre, The Robert Jones and Agnes Hunt Orthopaedic Hospital, Oswestry ; Spinal Studies, The Robert Jones and Agnes Hunt Orthopaedic Hospital, Oswestry ; Countess of Chester Hospital ; University of Chester ; University of Chester (Wiley, 2006-05-26)
      An increasing number of cell types, including peripheral blood mononuclear cells (PBMCs), have been demonstrated to release heat shock proteins (Hsps). This paper investigates further the hypothesis that Hsps are danger signals. PBMCs and Jurkat cells released Hsp70 (0·22 and 0·7 ng/106 cells, respectively) into medium over 24 h at 37°C. Release of Hsp70 was stimulated 10-fold by GroEL (P < 0·001) and more than threefold by lipopolysaccharide (LPS) (P < 0·001). Although Hsp60 could be detected in the medium of cells cultured at 37°C for 24 h, the low rates of release were due probably to cell damage. Significant release of Hsp60 was observed when Jurkat cells were exposed to GroEL (2·88 ng/106 cells) or LPS (1·40 ng/106 cells). The data are consistent with the hypothesis that Hsp70 and Hsp60 are part of a danger signalling cascade in response to bacterial infection.
    • Human placental oxygenation in late gestation: experimental and theoretical approaches

      Nye, Gareth; Ingram, Emma; Jenson, Oliver; Johnstone, Edward; Schneider, Henning; Lewis, Rohan; Chernyavsky, Igor; Brownbill, Paul; University of Manchester, University of Southampton, University of Bern (Wiley, 2018-01-26)
      The placenta is crucial for life. It is an ephemeral but complex organ acting as the barrier interface between maternal and fetal circulations, providing exchange of gases, nutrients, hormones, waste products and immunoglobulins. Many gaps exist in our understanding of the detailed placental structure and function, particularly in relation to oxygen handling and transfer in healthy and pathological states in utero. Measurements to understand oxygen transfer in vivo in the human are limited, with no general agreement on the most appropriate methods. An invasive method for measuring partial pressure of oxygen in the intervillous space through needle electrode insertion at the time of Caesarean sections has been reported. This allows for direct measurements in vivo whilst maintaining near normal placental conditions; however, there are practical and ethical implications in using this method for determination of placental oxygenation. Furthermore, oxygen levels are likely to be highly heterogeneous within the placenta. Emerging non-invasive techniques, such as MRI, and ex vivo research are capable of enhancing and improving current imaging methodology for placental villous structure and increase the precision of oxygen measurement within placental compartments. These techniques, in combination with mathematical modelling, have stimulated novel cross-disciplinary approaches that could advance our understanding of placental oxygenation and its metabolism in normal and pathological pregnancies, improving clinical treatment options and ultimately outcomes for the patient.
    • Individual, social, and environmental factors affecting salivary and fecal cortisol levels in captive pied tamarins (Saguinus bicolor)

      Wormell, Dominic; Smith, Tessa E.; Price, Eluned E.; Ahsmann, J.; Glendewar, G.; Hunt, J.; Coleman, Robert, C.; University of Chester (Wiley, 2019-08-01)
      Pied tamarins (Saguinus bicolor) are endangered New World primates, and in captivity appear to be very susceptible to stress. We measured cortisol in 214 saliva samples from 36 tamarins and in 227 fecal samples from 27 tamarins, and investigated the effects of age, sex, pregnancy, rearing history, social status, weight, group composition, and enclosure type using generalized linear mixed models. There was no effect of age on either fecal or salivary cortisol levels. Female pied tamarins in late pregnancy had higher fecal cortisol levels than those in early pregnancy, or nonpregnant females, but there was no effect of pregnancy on salivary cortisol. Females had higher salivary cortisol levels than males, but there was no effect of rearing history. However, for fecal cortisol, there was an interaction between sex and rearing history. Hand‐reared tamarins overall had higher fecal cortisol levels, but while male parent‐reared tamarins had higher levels than females who were parent‐ reared, the reverse was true for hand‐reared individuals. There was a trend towards lower fecal cortisol levels in subordinate individuals, but no effect of status on salivary cortisol. Fecal but not salivary cortisol levels declined with increasing weight. We found little effect of group composition on cortisol levels in either saliva or feces, suggesting that as long as tamarins are housed socially, the nature of the group is of less importance. However, animals in off‐show enclosures had higher salivary and fecal cortisol levels than individuals housed on‐show. We suggest that large on‐show enclosures with permanent access to off‐exhibit areas may compensate for the effects of visitor disturbance, and a larger number of tamarins of the same species housed close together may explain the higher cortisol levels found in tamarins living in off‐show accommodation, but further research is needed.
    • An investigation to determine the nutritional adequacy and individuals experience of a very low fat diet used to treat type V hypertriglyceridaemia

      Whitfield-Brown, Louisa M.; Hamer, O.; Ellahi, Basma; Burden, Sorrel; Durrington, Paul; University of Chester ; Manchester Royal Infirmary ; University of Chester ; Manchester Royal Infirmary ; University of Manchester (Wiley, 2009-05-15)
      This article discusses a study of eight patients with type V hypertriglyceridaemia on a low fat diet. The nutritional adequact of the diet and the barriers and enablers to adherence were analysed.
    • Is Wounding Aggression in Zoo-housed Chimpanzees and Ring-tailed Lemurs related to Zoo Visitor Numbers?

      Hosey, Geoff; Melfi, Vicky; Formella, Isabel; Ward, Samantha J.; Tokarski, Marina; Brunger, Dave; Brice, Sara; Hill, Sonya P.; University of Bolton; Taronga Zoo; South Lakes Wild Animal Park; Nottingham Trent University; Chester Zoo; University of Chester (Wiley, 2016-02-29)
      Chimpanzees in laboratory colonies experience more wounds on week days than on weekends, which has been attributed to the increased number of people present during the week; thus the presence of more people was interpreted as stressful. If this were also true for primates in zoos, where high human presence is a regular feature, this would clearly be of concern. Here we examine wounding rates in two primate species (chimpanzees Pan troglodytes and ring-tailed lemurs Lemur catta) at three different zoos, to determine whether they correlate with mean number of visitors to the zoo. Wounding data were obtained from zoo electronic record keeping system (ZIMS™). The pattern of wounds did not correlate with mean gate numbers for those days for either species in any group. We conclude that there is no evidence that high visitor numbers result in increased woundings in these two species when housed in zoos.
    • Local adaptation with high gene flow: temperature parameters drive adaptation to altitude in the common frog (Rana temporaria)

      Muir, Anna P.; Biek, Roman; Thomas, R.; Mable, Barbara K.; University of Chester; University of Glasgow; Royal Zoological Society of Scotland (Wiley, 2014-01-20)
      Both environmental and genetic influences can result in phenotypic variation. Quantifying the relative contributions of local adaptation and phenotypic plasticity to phenotypes is key to understanding the effect of environmental variation on populations. Identifying the selective pressures that drive divergence is an important, but often lacking, next step. High gene flow between high- and low-altitude common frog (Rana temporaria) breeding sites has previously been demonstrated in Scotland. The aim of this study was to assess whether local adaptation occurs in the face of high gene flow and to identify potential environmental selection pressures that drive adaptation. Phenotypic variation in larval traits was quantified in R. temporaria from paired high- and low-altitude sites using three common temperature treatments. Local adaptation was assessed using QST -FST analyses, and quantitative phenotypic divergence was related to environmental parameters using Mantel tests. Although evidence of local adaptation was found for all traits measured, only variation in larval period and growth rate was consistent with adaptation to altitude. Moreover, this was only evident in the three mountains with the highest high-altitude sites. This variation was correlated with mean summer and winter temperatures, suggesting that temperature parameters are potentially strong selective pressures maintaining local adaptation, despite high gene flow.
    • The low-risk perception of developing type 2 diabetes among women with a previous history of gestational diabetes: a qualitative study

      Sharma, Manisha; Purewal, Tejpal Singh; Fallows, Stephen; Kennedy, Lynne; Edge Hill University; Royal Liverpool Hospital; University of Chester (Wiley, 2019-02-12)
      We conducted a qualitative study to explore the risk perceptions, health beliefs and behaviours of women with a previous history of gestational diabetes mellitus (GDM). Women aged between 18 to 40 years (at the time of pregnancy) with a previous history of GDM, registered at The Royal Liverpool University Hospital, United Kingdom, participated in individual, semi-structured, face-to-face interviews. Qualitative data from seven participants were collected until data saturation and were analysed by thematic analysis. Participants had a low-risk perception of the future risk of developing diabetes. Some believed that their risk was the same as that of any other woman without a history of GDM, and some other participants were not aware of the risk at all and perceived GDM as a temporary health condition with no long-term risks. Participants showed some understanding of a healthy lifestyle in general. However, most of the information was self-acquired by participants and not linked to the future risk of developing diabetes. The findings of this research also indicated a contrast between the high perception of the immediate risks of complications during the pregnancy and low long-term risk of developing diabetes after pregnancy associated with GDM. Participants received healthy lifestyle advice during their pregnancy, but none of them reported involvement in any postnatal health education, intervention or counselling as recommended by 2008 and 2014 NICE guidelines. The low-risk perception impedes positive health behaviour required to overcome the barriers against a healthy lifestyle. This was a small research project but the findings warrant scope for more research in this field. A larger study might promote the development of a well-structured, long-term follow-up health intervention programme, incorporating a reminder system for annual diabetes screenings to improve the risk perception and reduce the risk for the development of type 2 diabetes in this population.
    • Magnetic nanoparticle-mediated gene delivery to two- and three-dimensional neural stem cell cultures: magnet-assisted transfection and multifection approaches to enhance outcomes

      Pickard, Mark R.; Adams, Christopher F.; Chari, Divya M.; University of Chester; Keele University (Wiley, 2017-02-02)
      Neural stem cells (NSCs) have high translational potential in transplantation therapies for neural repair. Enhancement of their therapeutic capacity by genetic engineering is an important goal for regenerative neurology. Magnetic nanoparticles (MNPs) are major non-viral vectors for safe bioengineering of NSCs, offering critical translational benefits over viral vectors, including safety, scalability, and ease of use. This unit describes protocols for the production of suspension (neurosphere) and adherent (monolayer) murine NSC cultures. Genetic engineering of NSCs with MNPs and the application of 'magnetofection' (magnetic fields) or 'multifection' (repeat transfection) approaches to enhance gene delivery are described. Magnetofection of monolayer cultures achieves optimal transfection, but neurospheres offer key advantages for neural graft survival post-transplantation. A protocol is presented which allows the advantageous features of each approach to be combined into a single procedure for transplantation. The adaptation of these protocols for other MNP preparations is considered, with emphasis on the evaluation of procedural safety.