Avian articular cartilage: effects of age, genotype and disease

The degradation of articular cartilage, causing degenerative joint disease (DJD), is a documented cause of lameness in broiler strain fowl, which is a major welfare problem. Broiler strain fowl are both heavier and more susceptible to DJD than laying strain fowl. In this thesis the biochemical and morphological basis for this susceptibility has been investigated, particular attention has been paid to the effects of body weight and genotype on avian articular cartilageArticular cartilage from three distinct sites was analysed. Samples from mature broiler strain females, susceptible to DJD, had higher hydration and uronic acid content than age matched, non -susceptible, laying strain fowl. In addition to these biochemical features (also seen in mammalian DJD), broiler strain fowl exhibited DJD histopathology including cartilage thinning and chondrocyte cluster formation. In general, both strains at one day old showed no significant biochemical differences between the articular cartilage sites sampled. However the cartilage from the broiler strain distal tibiotarsus (DTT) at one day old appeared to be biochemically and morphologically distinct from the cartilage of the layer strain DTT and from the other broiler strain joint surface of the same age.Production of an infectious arthritis by inoculation of mycoplasma into the tibiotarsal joint of broiler strain fowl resulted in variable biochemistry of articular cartilage form this joint. However in the non -injected, contralateral joint, sampled from the DTT, there was an increase in hydration and uronic acid content which is dependent upon the degree of lameness. This indicates the importance of in vivo loading in the biochemical composition of avian articular cartilage.Broiler strain birds fed ad libitum, feed restricted and J -line (wild type) were surveyed over the course of one year. Only the ad libitum fed birds developed overt DJD, which suggests that the mass of the bird, and not an overriding genetic element, is the major cause of the susceptibility of broiler strain fowl to DJD. The joint surface which presented the first and most severe signs indicative of early cartilage degeneration was the DTT. The biochemical results obtained from articular cartilage samples of the three groups include hydration, uronic acid, DNA and hydroxyproline content. Histology of the samples was assessed using haematoxylin and eosin stained sections. Proteoglycan content was investigated further by using selected samples for sulphated glycosaminoglycan assays and staining selected sections with Toludine blue and Safranin O. Samples from diseased and non -diseased groups were assayed for pyridinium in order to investigate the role of mature collagen crosslinking in DJD.The exact role of load and the metabolic consequences of body weight in the development of DJD is controversial. A method for artificially loading non -obese birds has been developed. Loading feed restricted broiler strain fowl with an additional 10% of their body weight over a three week period initiated a response including significant biochemical changes in the cartilage of the DTT. These results again emphasise the importance of load on the articular cartilage and the susceptibility of the DTT to DJD

The Lived Experience of Refractory Breathlessness

Introduction: Refractory breathlessness is breathlessness at rest or on minimal exertion that persists chronically even with optimal treatment of the underlying cause. Despite its burden and prevalence globally, symptoms remain under-treated and under-researched and those affected struggle daily with the condition. Aim: The aim of the study was to gain a greater understanding of the lived experience of refractory breathlessness. Also, it was anticipated that a deeper insight into its impacts on daily activities would allow more effective occupational therapy interventions to be developed. Method: A qualitative phenomenological approach using NVivo software, with one-on-one, semi-structured interviews and themes established through thematic analysis. The study recruited consecutive patients until thematic saturation was reached. Participants were recruited from a Sydney-based outpatient Breathlessness Clinic at a palliative care hospital. Results: 6 participants were interviewed and 4 main themes identified. 1) Living on the Edge, referring to the extreme feeling of breathlessness experienced by patients; 2) Social Needs, describing the significance of social support; 3) Reduced to the Basics, referring to the effect breathlessness has on basic daily tasks, and 4) Sources of Security, which describes the measures participants take to protect themselves from their own breathlessness. For each of the four themes, three to five subthemes were also identified through the analysis. Conclusion: Of the 4 themes identified in this study, number 3 and 4 may represent the best targets for occupational therapy interventions. Patients noted the importance of functional participation for improved engagement and quality of life. There is scope to develop specific interventions for self management and equipment prescription given lack of published literature in this area

Development of an incompressible Navier-Stokes solver for moving body problems using an overset meshing approach

An overset meshing approach is an effective method of simulating fluid flow involving multiple moving bodies. It consists of minor meshes representing solid objects, which overlap a Cartesian background grid, allowing bodies to move arbitrarily whilst retaining communication between grids. However, a hole beneath each overlapping mesh must be cut from the background grid, leaving a small overlap. Current hole-cutting methods tend to be complex with some requiring extensive user knowledge and input. Since the hole must be re-cut regularly for moving body problems, it can become very time-consuming. An original approach for performing a hole-cut has been implemented by employing the Cartesian cut-cell method. This method would ordinarily be used to cut the boundary of a solid object from a single Cartesian grid, as an alternative to the overset grids approach. Thus, the treatment of cut-cells has been modified for its new purpose of hole-cutting. The cut-cell method is already a well-established technique for cutting a Cartesian grid, and is fully automated. It has not been used for hole-cutting previously within the literature and offers a very different hole-cut to existing techniques; It cuts though cells rather than around them, simplifying the cutting process and providing a smooth cut. This approach has been applied to an incompressible Navier-Stokes solver for viscous single fluid flow. Unstructured, triangular minor meshes are used due to their ability to represent complex geometries accurately. An explicit time integration method is used on these minor meshes, but an implicit integration method is implemented on the Cartesian background mesh. This new hybrid of integration methods was found to significantly reduce the CPU time in comparison to using a fully explicit method. The solver has been validated using benchmark tests, including a lid driven cavity and flow past a stationary/oscillating cylinder. The results obtained were found to be in good quantitative agreement with published numerical results. The solver was developed for 2-phase flow problems. However, during the initial validation test, convergence issues were encountered, which meant a sufficient solution could not be obtained

Understanding forced marriage in Scotland

Executive summary In September 2015, the Scottish Government commissioned a 10 month study to better understand forced marriage in Scotland. This is the first forced marriage study that focuses exclusively on Scotland. The study had three research questions: What is the level and profile of service use relating to forced marriage in Scotland? How are services responding to forced marriage in Scotland? What is the impact of the interventions for forced marriage in Scotland? Methods The study focussed on six areas: Aberdeen, Dundee, East Renfrewshire, Edinburgh, Glasgow and Highland. An analysis of forced marriage policy in each of the six case study areas was undertaken, together with interviews with Protection Leads. One-to-one semi-structured interviews were carried out with a range of professionals including police officers, social workers, legal professionals and staff from Women's Aid organisations and other third sector organisations. Eight in-depth interviews were also conducted with survivors[1] of forced marriage. Additionally, a survey was distributed to 293 organisations - schools and women's organisations in the six study areas, and Women's Aid, minority ethnic and other support organisations throughout Scotland. Key findings and recommendations Level and profile of service use Between 2011 and 2014, there were 191 cases[2] of forced marriage reported by survey respondents, with a fairly even spread across the years. Cases tended to involve young, female South Asian victims being threatened or coerced into marriage largely by their parents and extended families. Age and ethnicity were unknown for around a quarter and a fifth of cases respectively, indicating that better recording of cases is necessary. Of the cases where age was known, the majority of victims were aged 18-25, with under 18s representing around a quarter of cases and under 16s around 1 in 10 of cases. Victims were mainly from Pakistani backgrounds (more than half of cases where ethnicity was known), followed by 'other ethnicity', Indian and Black African. Just over a half of referrals came to the attention of services through referral from other organisations, with under a third self-referrals and the remainder referred by friends or family. The interviews with the eight survivors of forced marriage echo the survey findings. The age range of when the forced marriage occurred was from 14-25. For five of the eight survivors the forced marriage was contracted with one survivor being forced into marriage twice. For two of the eight women, the experience of forced marriage was recent or ongoing (within the last two years). All the survivors interviewed were South Asian. All the women received help only when they came to Scotland - whether from abroad or another of the UK nations. Responding to forced marriage in Scotland Of the professionals interviewed, the majority had little direct experience of dealing with forced marriage but showed a sound understanding of forced marriage. However, those with direct experience of supporting victims of forced marriage had a deeper and more insightful understanding of forced marriage as a process rather than an 'event'. This was reflected at a policy level where there were differing levels of ownership and maturity of forced marriage policy, and where learning from forced marriage cases was identified as a means of improving responsiveness. Most professionals were aware that forced marriage took place in a range of communities, but it was seen as largely affecting South Asian communities in Scotland. This perception was confirmed by the survey findings, which found that the majority of cases where ethnicity was recorded related to South Asian communities. In both the policy analysis element of the study and in interviews with professionals, participants drew heavily from the Scottish Government's multi-agency guidance on forced marriage. A wide range of interventions are currently offered to support victims of forced marriage, most commonly one-to-one support, central to which is risk assessment and safety planning. Person-centred approaches were often used, and the need for therapeutic and practical support was also highlighted. Two third sector agencies interviewed offered mediation (and they were of the opinion that this was an effective intervention), despite the fact that Scottish multi-agency guidance stipulates that mediation should not be attempted in forced marriage cases. In terms of the outcome of support offered, nearly half of survey respondents reported that support offered was effective, but a further third were unsure whether the support offered was effective. Reported barriers to responding to forced marriage included 'race anxiety'; a need for more robust local authority procedures for supporting adult victims of forced marriage who have capacity (i.e. who do not meet the criteria to trigger access to adult protection); a need for further regular training for professionals; and a need for increased community education/public awareness on forced marriage. Multi-agency working was discussed both as very positive but also as a barrier - due to competing priorities and processes, with some agencies having more power than others to intervene and not always utilising the expertise available via specialists. A lack of training and learning opportunities on forced marriage was also identified as a barrier to responding effectively to forced marriage, despite forced marriage training being widely available. The survey results, policy analysis in some areas and interviews with some professionals suggest that a number of agencies and areas do not consider forced marriage a relevant issue for their organisation. This indicates that even with the provision of increased training on forced marriage, it might be challenging to recruit professionals to attend. Community education was thought to be a way to raise awareness within communities to increase reporting, but this intervention can also serve as a preventive measure. Targeted interventions for different sections of communities e.g. young people and elders would need to be developed. Although there was wide publicity about the forced marriage legislation, a lack of public awareness of forced marriage was also cited as a major barrier to increasing reporting and providing support for victims. This links with survivors' accounts that they sought help from family and friends and feared contacting agencies due to perceived confidentiality issues, family pressure, uncertainty about the appropriateness of response, and uncertainty about whether what had or was happening to them warranted agency involvement. There are examples of good practice at both a policy and practice level, including meaningful connections between child and adult protection leads and violence against women leads in some areas, proactive learning, person-centred support and in-depth expertise on forced marriage. Survivors reported that they had received excellent support from third sector organisations, but their accounts of other agencies were more mixed. Impact of interventions There was widespread support for civil remedies for dealing with Forced Marriage, from professionals interviewed and survivors of forced marriage. A number of issues were identified with regard to the implementation of civil remedies, relating to: a lack of consensus about what constitutes 'sufficient' evidence to justify granting a Forced Marriage Protection Order the onus of responsibility being placed on the victim, particularly where the victim is an adult who does not meet the criteria for accessing adult protection confidentiality of the victim not always being maintained In relation to criminalisation, most professionals interviewed thought it sent a strong message to the public that forced marriage was unacceptable in Scotland. However, a number of professionals (including some who supported criminalisation) also raised concerns about the potential for forced marriage to 'go underground', as victims would not wish to criminalise their families. It is too early to say whether this concern is justified. All survivors welcomed legal protection, but most were not supportive of criminalisation. However, one survivor who had pursued an FMPO stated that criminal procedures should be used, but only as a last resort. The impact of forced marriage on survivors of forced marriage included suicidal ideation, self-harm, eating disorders and other mental health problems. Survivors were frequently denied educational opportunities, impacting on their future careers and earning capacity. On a more positive note, survivors' experiences of forced marriage had engendered within them more liberal attitudes to parenting than those that they had experienced from their own parents. Survivors' experiences of statutory service response, including recent responses in Scotland, although positive in places, was patchy. However, it should be noted that some of the women were reflecting on historic cases stretching back a couple of decades and, at times, referring to experiences that happened outside Scotland. Most of the women had had some contact with third sector specialist women's support organisations, and all reported this as an extremely positive experience, although it should be noted that most survivors were recruited through their engagement with the women's sector. For some of the women, this was the first time they had told their story; for all of them it was the first time they had received support. Recommendations 1. Develop an innovative programme of further public/community awareness-raising activity, to prevent forced marriage and to encourage increased reporting 2. Develop further regular training on forced marriage for a range of professionals (including teachers, social workers, police officers, legal professionals and mental health practitioners), and ensure appropriate staff attend and the learning is cascaded and applied 3. Support the continued development of specialist women's sector organisations 4. Support the development of forced marriage policy in local authority areas, in order to increase ownership and consistency of approach at a local policy level throughout Scotland 5. Ensure that therapeutic and practical support is available to victims of forced marriage 6. Evaluate forced marriage interventions to develop better understandings of what types of forced marriage interventions work for whom 7. Address the issues the research identified in relation to implementing forced marriage legislation 8. Strengthen the statutory guidance of the Forced Marriage etc. (Protection and Jurisdiction) (Scotland) Act (2011) to make explicit local authorities' obligations to act in all cases of Forced Marriage 9. Improve record keeping of cases of forced marriage, as information about cases is key to developing learning and for future policy developmen

RCAN1 regulates vesicle recycling and quantal release kinetics via effects on calcineurin activity

Author version made available in accordance with the publisher's policy.We have previously shown that Regulator of Calcineurin 1 (RCAN1) regulates multiple stages of vesicle exocytosis. However, the mechanisms by which RCAN1 affects secretory vesicle exocytosis and quantal release kinetics remain unknown. Here we use carbon fiber amperometry to detect exocytosis from chromaffin cells and identify these underlying mechanisms. We observe reduced exocytosis with repeated stimulations in chromaffin cells overexpressing RCAN1 (RCAN1ox), but not in wild type (WT) cells, indicating a negative effect of RCAN1 on vesicle recycling and endocytosis. Acute exposure to calcineurin inhibitors, cyclosporine A and FK-506, replicates this effect in WT cells but has no additional effect in RCAN1ox cells. When we chronically expose WT cells to cyclosporine A and FK-506 we find that catecholamine release per vesicle and pre-spike foot (PSF) signal parameters are decreased, similar to that in RCAN1ox cells. Inhibiting calcineurin activity in RCAN1ox cells has no additional effect on the amount of catecholamine release per vesicle but further reduces PSF signal parameters. Electron microscopy studies indicate these changes are not due to altered vesicle number or distribution in RCAN1ox cells but reduced vesicle release may be cause by decreased vesicle and dense core size in RCAN1ox cells. Thus, our results indicate that RCAN1 may negatively affects vesicle recycling and quantal release kinetics via the inhibition of calcineurin activity

Incorporating a mucosal environment in a dynamic gut model results in a more representative colonization by lactobacilli

To avoid detrimental interactions with intestinal microbes, the human epithelium is covered with a protective mucus layer that traps host defence molecules. Microbial properties such as adhesion to mucus further result in a unique mucosal microbiota with a great potential to interact with the host. As mucosal microbes are difficult to study in vivo, we incorporated mucin-covered microcosms in a dynamic in vitro gut model, the simulator of the human intestinal microbial ecosystem (SHIME). We assessed the importance of the mucosal environment in this M-SHIME (mucosal-SHIME) for the colonization of lactobacilli, a group for which the mucus binding domain was recently discovered. Whereas the two dominant resident Lactobacilli, Lactobacillus mucosae and Pediococcus acidilactici, were both present in the lumen, L. mucosae was strongly enriched in mucus. As a possible explanation, the gene encoding a mucus binding (mub) protein was detected by PCR in L. mucosae. Also the strongly adherent Lactobacillus rhamnosus GG (LGG) specifically colonized mucus upon inoculation. Short-term assays confirmed the strong mucin-binding of both L. mucosae and LGG compared with P. acidilactici. The mucosal environment also increased long-term colonization of L. mucosae and enhanced its stability upon antibiotic treatment (tetracycline, amoxicillin and ciprofloxacin). Incorporating a mucosal environment thus allowed colonization of specific microbes such as L. mucosae and LGG, in correspondence with the in vivo situation. This may lead to more in vivo-like microbial communities in such dynamic, long-term in vitro simulations and allow the study of the unique mucosal microbiota in health and disease

East Weddell Sea echinoids from the JR275 expedition

Information regarding the echinoids in this dataset is based on the Agassiz Trawl (AGT) and epibenthic sledge (EBS) samples collected during the British Antarctic Survey cruise JR275 on the RRS James Clark Ross in the austral summer 2012. A total of 56 (1 at the South Orkneys and 55 in the Eastern Weddell Sea) Agassiz Trawl and 18 (2 at the South Orkneys and 16 in the Eastern Weddell Sea) epibenthic sledge deployments were performed at depths ranging from ~280 to ~2060 m. This presents a unique collection for the Antarctic benthic biodiversity assessment of an important group of benthic invertebrates. In total 487 specimens belonging to six families, 15 genera, and 22 morphospecies were collected. The species richness per station varied between one and six. Total species richness represents 27% of the 82 echinoid species ever recorded in the Southern Ocean (David et al. 2005b, Pierrat et al. 2012, Saucède et al. 2014). The Cidaridae (sub-family Ctenocidarinae) and Schizasteridae are the two most speciose families in the dataset. They comprise seven and nine species respectively. This is illustrative of the overall pattern of echinoid diversity in the Southern Ocean where 65% of Antarctic species belong to the families Schizasteridae and Cidaridae (Pierrat et al. 2012)