Lipid changes within the epidermis of living skin equivalents observed across a time-course by MALDI-MS imaging and profiling

© 2015 Mitchell et al. Abstract Background: Mass spectrometry imaging (MSI) is a powerful tool for the study of intact tissue sections. Here, its application to the study of the distribution of lipids in sections of reconstructed living skin equivalents during their development and maturation is described. Methods: Living skin equivalent (LSE) samples were obtained at 14 days development, re-suspended in maintenance medium and incubated for 24 h after delivery. The medium was then changed, the LSE re-incubated and samples taken at 4, 6 and 24 h time points. Mass spectra and mass spectral images were recorded from 12 μm sections of the LSE taken at each time point for comparison using matrix assisted laser desorption ionisation mass spectrometry. Results: A large number of lipid species were identified in the LSE via accurate mass-measurement MS and MSMS experiments carried out directly on the tissue sections. MS images acquired at a spatial resolution of 50 μm × 50 μm showed the distribution of identified lipids within the developing LSE and changes in their distribution with time. In particular development of an epidermal layer was observable as a compaction of the distribution of phosphatidylcholine species. Conclusions: MSI can be used to study changes in lipid composition in LSE. Determination of the changes in lipid distribution during the maturation of the LSE will assist in the identification of treatment responses in future investigations

Mental health help-seeking behaviours in young adults

There is clinical and research consensus that significant cognitive, social, emotional development and adjustment to physical changes occurs during young adulthood, in the period between 18 and 24 years1. Whilst three quarters of psychiatric disorders in adults emerge before the age of 25 years , a European study, comparing access to mental healthcare by age bands, reported that 18-24 year old participants were least likely to get care for mental health problems 2. In the 2016 UK National Confidential Enquiry into Suicide in Children and Young People, 43% of people under the age of 25 who died had no known prior contact with any agencies3. Understanding the risk factors and triggers for mental health problems in young adults is crucial, however we also need to know more about how young adults seek help, if we wish to improve the quality and outcomes of mental healthcare. Early interventions may improve the prognosis of primary mental health disorders in young adults and reduce the risk of chronicity and progression to more severe secondary disorders, but research led innovation in mental health care is also hampered by delayed diagnostic assessment and care

Special research paper

A review of the literature on therapist variables in counselling and therapy; a comparison of the responses of two therapists who conducted one interview each with the same clien

Global Dimension of Polynomial Rings in Partially Commuting Variables

For any free partially commutative monoid $M(E,I)$, we compute the global dimension of the category of $M(E,I)$-objects in an Abelian category with exact coproducts. As a corollary, we generalize Hilbert's Syzygy Theorem to polynomial rings in partially commuting variables.Comment: 11 pages, 2 figure

Interpolatory methods for H∞\mathcal{H}_\infty model reduction of multi-input/multi-output systems

We develop here a computationally effective approach for producing high-quality $\mathcal{H}_\infty$-approximations to large scale linear dynamical systems having multiple inputs and multiple outputs (MIMO). We extend an approach for $\mathcal{H}_\infty$ model reduction introduced by Flagg, Beattie, and Gugercin for the single-input/single-output (SISO) setting, which combined ideas originating in interpolatory $\mathcal{H}_2$-optimal model reduction with complex Chebyshev approximation. Retaining this framework, our approach to the MIMO problem has its principal computational cost dominated by (sparse) linear solves, and so it can remain an effective strategy in many large-scale settings. We are able to avoid computationally demanding $\mathcal{H}_\infty$ norm calculations that are normally required to monitor progress within each optimization cycle through the use of "data-driven" rational approximations that are built upon previously computed function samples. Numerical examples are included that illustrate our approach. We produce high fidelity reduced models having consistently better $\mathcal{H}_\infty$ performance than models produced via balanced truncation; these models often are as good as (and occasionally better than) models produced using optimal Hankel norm approximation as well. In all cases considered, the method described here produces reduced models at far lower cost than is possible with either balanced truncation or optimal Hankel norm approximation

MAPK phosphorylation of connexin 43 promotes binding of cyclin E and smooth muscle cell proliferation

<p>Rationale: Dedifferentiation of vascular smooth muscle cells (VSMC) leading to a proliferative cell phenotype significantly contributes to the development of atherosclerosis. Mitogen-activated protein kinase (MAPK) phosphorylation of proteins including connexin 43 (Cx43) has been associated with VSMC proliferation in atherosclerosis.</p> <p>Objective: To investigate whether MAPK phosphorylation of Cx43 is directly involved in VSMC proliferation.</p> <p>Methods and Results: We show in vivo that MAPK-phosphorylated Cx43 forms complexes with the cell cycle control proteins cyclin E and cyclin-dependent kinase 2 (CDK2) in carotids of apolipoprotein-E receptor null (ApoE−/−) mice and in C57Bl/6 mice treated with platelet-derived growth factor–BB (PDGF). We tested the involvement of Cx43 MAPK phosphorylation in vitro using constructs for full-length Cx43 (Cx43) or the Cx43 C-terminus (Cx43CT) and produced null phosphorylation Ser>Ala (Cx43MK4A/Cx43CTMK4A) and phospho-mimetic Ser>Asp (Cx43MK4D/Cx43CTMK4D) mutations. Coimmunoprecipitation studies in primary VSMC isolated from Cx43 wild-type (Cx43+/+) and Cx43 null (Cx43−/−) mice and analytic size exclusion studies of purified proteins identify that interactions between cyclin E and Cx43 requires Cx43 MAPK phosphorylation. We further demonstrate that Cx43 MAPK phosphorylation is required for PDGF-mediated VSMC proliferation. Finally, using a novel knock-in mouse containing Cx43-MK4A mutation, we show in vivo that interactions between Cx43 and cyclin E are lost and VSMC proliferation does not occur after treatment of carotids with PDGF and that neointima formation is significantly reduced in carotids after injury.</p> <p>Conclusions: We identify MAPK-phosphorylated Cx43 as a novel interacting partner of cyclin E in VSMC and show that this interaction is critical for VSMC proliferation. This novel interaction may be important in the development of atherosclerotic lesions.</p&gt

Older users’ requirements for interactive television

Older users’ requirements for interactive televisio

Respiratory health screening for opiate misusers in a specialist community clinic: a mixed-methods pilot study, with integrated staff and service user feedback.

OBJECTIVES: Increased rates of illicit drug inhalation are thought to expose opiate misusers (OMUs) to an enhanced risk of respiratory health problems. This pilot study aimed to determine the feasibility of undertaking respiratory screening of OMUs in a community clinic. SETTING: Single-centre UK community substance misuse clinic. PARTICIPANTS: All clinic attendees receiving treatment for opiate misuse were eligible to participate. 36 participants (mean age=37) were recruited over a 5-week period. The sample included 26 males and 10 females. OUTCOME MEASURES: Spirometry without bronchodilation; health related quality of life EQ-5D-3L; Asthma Control Test; Mini Asthma Quality of Life; Clinical COPD Questionnaire and the Treatment Outcome Profile were used to assess the respiratory health of participants. Findings were discussed with staff and service users in 2 patient and public involvement events and feedback was analysed thematically. RESULTS: 34 participants reported that they had smoked heroin. 8 participants diagnosed with asthma, scored under 13 on the Asthma Control Test, suggesting poorly controlled asthma. Participants (n=28), without a diagnosis of asthma completed the Lung Function Questionnaire. Of these, 79% produced scores under 18, indicating symptoms associated with the development of chronic obstructive pulmonary disease. Spirometry showed 14% of all participants had forced expiratory volume in 1 s/forced vital capacity <0.7 (without bronchodilator), indicating potential obstructive lung disease. Feedback from service users and staff suggested a willingness and capacity to deliver respiratory health screening programmes. Insight towards the difficulties service users have in accessing services and the burden of respiratory health was also provided. CONCLUSIONS: It is feasible to undertake respiratory health screening of OMUs in a community clinic. Larger screening studies are warranted to determine the prevalence of respiratory health problems in this population. Research regarding asthma medicines adherence and access to healthcare among OMUs is also required