Vitamin D binding protein isoforms as candidate predictors of disease extension in childhood arthritis

Introduction: Juvenile idiopathic arthritis (JIA) comprises a poorly understood group of chronic autoimmune diseases with variable clinical outcomes. We investigated whether the synovial fluid (SF) proteome could distinguish a subset of patients in whom disease extends to affect a large number of joints.

Methods: SF samples from 57 patients were obtained around time of initial diagnosis of JIA, labeled with Cy dyes and separated by two-dimensional electrophoresis. Multivariate analyses were used to isolate a panel of proteins which distinguish patient subgroups. Proteins were identified using MALDI-TOF mass spectrometry with expression verified by immunochemical methods. Protein glycosylation status was confirmed by hydrophilic interaction liquid chromatography.

Results: A truncated isoform of vitamin D binding protein (VDBP) is present at significantly reduced levels in the SF of oligoarticular patients at risk of disease extension, relative to other subgroups (p < 0.05). Furthermore, sialylated forms of immunopurified synovial VDBP were significantly reduced in extended oligoarticular patients (p < 0.005).

Conclusion: Reduced conversion of VDBP to a macrophage activation factor may be used to stratify patients to determine risk of disease extension in JIA patients.

Development of a rapid low cost fluorescent biosensor for the detection of food contaminants

A prototype fluorescent based biosensor has been developed for the antibody based detection of food related contaminants. Its performance was characterised and showed a typical antibody binding signal of 200-2000 mV, a short term noise of 9.1 mV, and baseline slope of -0.016 mV/s over 4 h. Bulk signal detection repeatability (n=23) and reproducibility (n=3) were less than 2.4%CV. The biosensor detection unit was evaluated using two food related model systems proving its ability to monitor both binding using commercial products and inhibition through the development of an assay. This assay development potential was evaluated by observing the biosensor's performance whilst appraising several labelled antibody and glass slide configurations. The molecular interaction between biotin and an anti-biotin antibody was shown to be inhibited by 41% due to the presence of biotin in a sample. A food toxin (domoic acid) calibration curve was produced, with %CVs ranging from 2.7 to 7.8%, and a midpoint of approximately 17 ng/ml with further optimisation possible. The ultimate aim of this study was to demonstrate the working principles of this innovative biosensor as a potential portable tool with the opportunity of interchangeable assays. The biosensor design is applicable for the requirements of routine food contaminant analysis, with respect to performance, functionality and cost. (C) 2012 Elsevier B.V. All rights reserved.

Palaeoglacial and palaeoclimatic conditions in the NW Pacific, as revealed by a morphometric analysis of cirques upon the Kamchatka Peninsula

The distribution of glacial cirques upon the Kamchatka peninsula, Far Eastern Russia, is systematically mapped from satellite images and digital elevation model data. A total of 3,758 cirques are identified, 238 of which are occupied by active glaciers. The morphometry of the remaining 3,520 cirques is analysed. These cirques are found to show a very strong N bias in their azimuth (orientation), likely resulting from aspect-related variations in insolation. The strength of this N bias is considered to indicate that former glaciation upon the peninsula was often ‘marginal’, and mainly of cirque-type, with peaks extending little above regional equilibrium-line altitudes. This is supported by the fact that S and SE-facing cirques are the highest in the dataset, suggesting that glacier-cover was rarely sufficient to allow S and SE-facing glaciers to develop at low altitudes. The strength of these azimuth-related variations in cirque altitude is thought to reflect comparatively cloud-free conditions during former periods of glaciation. It is suggested that these characteristics, of marginal glaciation and comparatively cloud-free conditions, reflect the region’s former aridity, which was likely intensified at the global Last Glacial Maximum, and during earlier periods of ice advance, as a result of the development of negative pressure anomalies over the North Pacific (driven by the growth of the Laurentide Ice Sheet), combined with other factors, including an increase in the extent and duration of sea ice, a reduction in global sea levels, cooler sea surface temperatures, and the localised growth of mountain glaciers. There is published evidence to suggest extensive glaciation of the Kamchatka Peninsula at times during the Late Quaternary, yet the data presented here appears to suggest that such phases were comparatively short-lived, and that smaller cirque-type glaciers were generally more characteristic of the period.

Effect-based proteomic detection of growth promoter abuse

Unregulated growth promoter use in food-producing animals is an issue of concern both from food safety and animal welfare perspectives. However, the monitoring of such practices is analytically challenging due to the concerted actions of users to evade detection. Techniques based on the monitoring of biological responses to exogenous administrations have been proposed as more sensitive methods to identify treated animals. This study has, for the first time, profiled plasma proteome responses in bovine animals to treatment with nortestosterone decanoate and 17 beta-oestradiol benzoate, followed by dexamethasone administration. Two-dimensional fluorescence differential in-gel electrophoresis analysis revealed a series of hepatic and acute-phase proteins within plasma whose levels were up- or down-regulated within phases of the treatment regime. Surface plasmon resonance (SPR) immuno-assays were developed to quantify responses of identified protein markers during the experimental treatment study with a view to developing methods which can be used as screening tools for growth promoter abuse detection. SPR analysis demonstrated the potential for plasma proteins to be used as indicative measures of growth promoter administrations and concludes that the sensitivity and robustness of any detection approach based on plasma proteome analysis would benefit from examination of a range of proteins representative of diverse biological processes rather being reliant on specific individual markers.

HOXA/PBX3 knockdown impairs growth and sensitizes cytogenetically normal acute myeloid leukemia cells to chemotherapy

The cytogenetically normal subtype of acute myeloid leukemia (CN-AML) is associated with Intermediate risk which complicates therapeutic options. Lower overall HOX/TALE expression appears to correlate with more favorable prognosis/better response to treatment in some leukemias and solid cancer. The functional significance of the associated gene expression and response to chemotherapy is not known. Three independent microarray datasets obtained from large patient cohorts along with quantitative PCR validation was used to identify a four gene HOXA/TALE signature capable of prognostic stratification. Biochemical analysis was used to identify interactions between the four encoded proteins and targeted knockdown used to examine the functional importance of sustained expression of the signature in leukemia maintenance and response to chemotherapy. An eleven HOXA/TALE code identified in an Intermediate risk (n=315) compared to a Favourable group of patients (n=105) was reduced to a four gene signature of HOXA6, HOXA9, PBX3 and MEIS1 by iterative analysis of independent platforms. This signature maintained the Favorable/Intermediate risk partition and where applicable, correlated with overall survival in CN-AML. We further show that cell growth and function is dependent on maintained levels of these core genes and that direct targeting of HOXA/PBX3 sensitizes CN-AML cells to standard chemotherapy. Together the data support a key role for HOXA/TALE in CN-AML and demonstrate that targeting of clinically significant HOXA/PBX3 elements may provide therapeutic benefit to these patients.

End-of-Life care and people with intellectual disabilities: A multi-media educational resource

Aim This study aimed to develop and evaluate a multi-media educational resource in palliative and end-of-life care for specialist palliative care and intellectual disability services which promoted collaborative working. Methods: A mixed methods design involving three phases was used. Qualitative data were obtained from semi-structured interviews with a purposive sample of professionals (n=30) and family carers (n=5) and from two focus groups with people with intellectual disabilities (n=17). Data were content analysed as outlined y Newell and Burnard (2006). This identified training needs and issues, in end-of-life care for this population which were confirmed through quantitative data from services in a regional scoping study analysed using descriptive statistics. A DVD and manual were developed and evaluated with twelve professionals. Data were collected using a solicited diary, the Readiness for Inter-professional Learning Scale, Likert Scales and an evaluation questionnaire. Thematic analysis and descriptive statistics appropriate to data were used.Results: Findings suggest that this resource demonstrates the need for and benefits of partnership working and transferability of this learning to practice could address issues at end-of-life for people with intellectual disabilities. Conclusions: Findings of this study have importance for partnership working and service provision in end-of-life care for this population.

Gay men and intimate partner violence: a gender analysis

Though intimate partner violence (IPV) is predominately understood as a women’s health issue most often emerging within heterosexual relationships, there is increasing recognition of the existence of male victims of IPV. In this qualitative study we explored connections between masculinities and IPV among gay men. The findings show how recognising IPV was based on an array of participant experiences, including the emotional, physical and sexual abuse inflicted by their partner, which in turn led to three processes. Normalising and concealing violence referred to the participants’ complicity in accepting violence as part of their relationship and their reluctance to disclose that they were victims of IPV. Realising a way out included the participants’ understandings that the triggers for, and patterns of, IPV would best be quelled by leaving the relationship. Nurturing recovery detailed the strategies employed by participants to mend and sustain their wellbeing in the aftermath of leaving an abusive relationship. In terms of masculinities and men’s health research, the findings reveal the limits of idealising hegemonic masculinities and gender relations as heterosexual, while highlighting a plurality of gay masculinities and the need for IPV support services that bridge the divide between male and female as well as between homosexual and heterosexual.