Development of methods for the in-vitro assessment of the adipogenesis and immunogenicity of human multipotent mesenchymal stromal cells

Adult mesenchymal stromal cells (MSCs) have enormous potential in tissue engineering and regenerative medicine applications. MSC have also been reported to be immune-privileged. The aim of this study was to develop methods to test two related hypotheses. Firstly, that MSCs will not directly stimulate allogeneic lymphocytes in classical lymphocyte proliferation assays carried out over 5-7 days due to lack of expression of immune-regulatory molecules (CD40, CD80, CD86 and MHC Class-II) but may be capable of the stimulation of allogeneic lymphocytes when cultured over extended time periods that allow the indirect pathway of antigen presentation. Secondly, that following differentiation of MSCs into mature differentiated adipose cells, the cells would become immunogenic or increase their immunogenicity, as determined by the stimulation of allogeneic peripheral blood mononuclear cells in lymphocyte transformation assays. MSCs from 4 human bone marrow donors were isolated, expanded and characterised and shown to conform to the ISCT definition of MSC through FACs analysis of key antigen expression and trilineage differentiation. MSCs (from donors 1-3) were differentiated into adipocytes, and current methodology for assaying adipogenesis was evaluated. A new fluorescent microplate assay was developed using DAPI to normalise for cell number and Nile red to stain for intracellular lipids, to generate a ratio of adipogenesis. Peripheral blood mononuclear cells (PBMCs) from 6 volunteers were isolated, in-vitro assay conditions for performing lymphocyte transformation assays (LTAs) were optimised, and the duration of the assays extended to 16-21 days. It was shown that undifferentiated MSCs stimulated PBMCs in all except 2 out of 24 cases, with the highest stimulation typically post-day 7 with stimulation indices as high as 36 observed. It was shown that MSCs differentiated into adipocytes also stimulated PBMCs with higher stimulation indices observed in differentiated MSCs compared to the undifferentiated counterparts. It was concluded that MSC, whether undifferentiated or differentiated into adipose cells, are indeed capable of stimulating a proliferative response in allogeneic lymphocytes. Therefore, the use of allogeneic MSCs in the traditional engineering setting may be limited, and a cautious approach should be taken before allogeneic MSCs are used in the regeneration or creation of tissue replacements for clinical use

Mussels can both outweigh and interact with the effects of terrestrial to freshwater resource subsidies on littoral benthic communities.

Litterfall is an important resource subsidy for lake ecosystems that primarily accumulates in littoral zones. Bivalves are abundant within littoral zones and may modify the effects of terrestrial resource subsidies through trophic interactions and engineering their surrounding habitat. Leaf inputs to lakes and freshwater mussel abundances are changing throughout the boreal ecoregion so we set out to investigate how the co-occurring benthic community might respond. We set up an in situ mesocosm experiment in Ramsey Lake, Sudbury, ON, Canada. Mesocosms contained sediments of either 5% or 35% terrestrial organic matter (tOM), into which we placed mussels (Elliptio complanata) at differing densities (0, 0.4 and 2musselsm-2, with a sham mussel treatment at 0.4musselsm-2). Over one month we recorded the sediment chemistry (dissolved organic carbon, nitrogen and phosphorus), littoral organisms (benthic algae and zooplankton) and mussel growth. At high mussel densities we recorded a 90%, 80%, 45% and 40% reduction in phosphorus, dissolved organic carbon, nitrogen and benthic diatoms, respectively, whereas at low mussel densities we observed a 3-fold increase in zooplankton. We discuss that these results were caused by a combination of bioturbation and trophic interactions. Benthic diatom concentrations were also reduced by 20% in sediments of 35% tOM, likely due to shading and competition with bacteria. Mussel growth increased at high mussel densities but was offset at high tOM, likely due to the organic matter interfering with filter feeding. Our results suggest that mussels can alter the geochemical composition of sediments and abundances of associated littoral organisms, in some cases regardless of tOM quantity. Therefore, the dominant top-down control exerted by freshwater mussels may outweigh bottom-up effects of tOM additions. Generally, our study reveals the importance of considering dominant species when studying the effects of cross-ecosystem resource fluxes

Patient attitudes towards analgesia and their openness to non-pharmacological methods such as acupuncture in the emergency department

Aims: To investigate patient attitudes to analgesia, opioids and non-pharmacological analgesia including acupuncture, in the ED. Methods: ED patients with pain were surveyed regarding: pain scores, satisfaction, addiction concern, non-pharmacological methods of pain relief, and acupuncture. Data were analysed using logistic regression. Results: Of 196 adult patients, 52.8% were ‘very satisfied’ with analgesia. Most patients (84.7%) would accept non-pharmacological methods including acupuncture (68.9%) and 78.6% were not concerned about addiction. Satisfaction was associated with male gender, and ‘adequate analgesia’ but not with opioids. Conclusion: Most patients were generally satisfied with ED analgesia and were open to non-pharmacologic analgesia including acupuncture

Development of methods for the in-vitro assessment of the adipogenesis and immunogenicity of human multipotent mesenchymal stromal cells

Adult mesenchymal stromal cells (MSCs) have enormous potential in tissue engineering and regenerative medicine applications. MSC have also been reported to be immune-privileged. The aim of this study was to develop methods to test two related hypotheses. Firstly, that MSCs will not directly stimulate allogeneic lymphocytes in classical lymphocyte proliferation assays carried out over 5-7 days due to lack of expression of immune-regulatory molecules (CD40, CD80, CD86 and MHC Class-II) but may be capable of the stimulation of allogeneic lymphocytes when cultured over extended time periods that allow the indirect pathway of antigen presentation. Secondly, that following differentiation of MSCs into mature differentiated adipose cells, the cells would become immunogenic or increase their immunogenicity, as determined by the stimulation of allogeneic peripheral blood mononuclear cells in lymphocyte transformation assays. MSCs from 4 human bone marrow donors were isolated, expanded and characterised and shown to conform to the ISCT definition of MSC through FACs analysis of key antigen expression and trilineage differentiation. MSCs (from donors 1-3) were differentiated into adipocytes, and current methodology for assaying adipogenesis was evaluated. A new fluorescent microplate assay was developed using DAPI to normalise for cell number and Nile red to stain for intracellular lipids, to generate a ratio of adipogenesis. Peripheral blood mononuclear cells (PBMCs) from 6 volunteers were isolated, in-vitro assay conditions for performing lymphocyte transformation assays (LTAs) were optimised, and the duration of the assays extended to 16-21 days. It was shown that undifferentiated MSCs stimulated PBMCs in all except 2 out of 24 cases, with the highest stimulation typically post-day 7 with stimulation indices as high as 36 observed. It was shown that MSCs differentiated into adipocytes also stimulated PBMCs with higher stimulation indices observed in differentiated MSCs compared to the undifferentiated counterparts. It was concluded that MSC, whether undifferentiated or differentiated into adipose cells, are indeed capable of stimulating a proliferative response in allogeneic lymphocytes. Therefore, the use of allogeneic MSCs in the traditional engineering setting may be limited, and a cautious approach should be taken before allogeneic MSCs are used in the regeneration or creation of tissue replacements for clinical use.EThOS - Electronic Theses Online ServiceGBUnited Kingdo

Antenatally diagnosed ADPKD

[Extract] Advances in antenatal ultrasonographyhave substantially improved the counseling of pregnantwomen.1�4With this advancement comes a diagnosticdilemma in the approach to antenatal diagnoses of cystickidney disease. The differential diagnosis of enlargedcystic echogenic kidneys includes autosomal dominantpolycystic kidney disease (ADPKD), autosomal recessivepolycystic kidney disease, renal cysts and diabetessyndrome, and other syndromic disorders

Forecasting climate change impacts on plant populations over large spatial extents

Plant population models are powerful tools for predicting climate change impacts in one location, but are difficult to apply at landscape scales. We overcome this limitation by taking advantage of two recent advances: remotely sensed, species-specific estimates of plant cover and statistical models developed for spatiotemporal dynamics of animal populations. Using computationally efficient model reparameterizations, we fit a spatiotemporal population model to a 28-year time series of sagebrush (Artemisia spp.) percent cover over a 2.5 × 5 km landscape in southwestern Wyoming while formally accounting for spatial autocorrelation. We include interannual variation in precipitation and temperature as covariates in the model to investigate how climate affects the cover of sagebrush. We then use the model to forecast the future abundance of sagebrush at the landscape scale under projected climate change, generating spatially explicit estimates of sagebrush population trajectories that have, until now, been impossible to produce at this scale. Our broadscale and long-term predictions are rooted in small-scale and short-term population dynamics and provide an alternative to predictions offered by species distribution models that do not include population dynamics. Our approach, which combines several existing techniques in a novel way, demonstrates the use of remote sensing data to model population responses to environmental change that play out at spatial scales far greater than the traditional field study plot

Integrated approach to cardiovascular disease in people experiencing homelessness: a qualitative study

BACKGROUND: Homelessness is associated with an increased risk of cardiovascular disease (CVD), beyond impact of socioeconomic status. CVD is preventable and treatable, though barriers to interventions exist for people experiencing homelessness. Those with lived experience of homelessness and health professionals with relevant expertise can help to understand and address these barriers. OBJECTIVES: To understand, and make recommendations to improve, CVD care in homeless populations through lived and professional expertise. METHOD: Four focus groups were conducted in March-July 2019. Three groups included people currently or previously experiencing homelessness, each attended by a cardiologist (AB), a health services researcher (PB) and an 'expert by experience' (SB) who coordinated participants. One group included multidisciplinary health and social care professionals in and around London to explore solutions. PARTICIPANTS: The three groups included 16 men and 9 women, aged 20-60 years, of whom 24 were homeless and currently living in hostels, and 1 rough sleeper. At least 14 discussed sleeping rough at some point. RESULTS: Participants were aware of CVD risks and relevance of healthy habits but identified barriers to prevention and health access, starting with disorientation affecting planning and self-care, lack of facilities for food, hygiene and exercise, and experiences of discrimination. CONCLUSIONS: CVD care for those experiencing homelessness should account for fundamental problems of the environment, be codesigned with service users and cover key principles: flexibility, public and staff education, integration of support and advocacy for health service rights

‘I Didn’t Have the Language Then’—A Qualitative Examination of Terminology in the Development of Non-Binary Identities

Introduction: Identities that lie outside of exclusively male and female, such as non-binary and genderqueer, have become increasingly more prevalent and visible within recent years. However, to date, the role of terminology in the development of such gender identities has been under-researched. This study aims to: (1) Examine what role terminology plays in coming to identify as non-binary. (2) Explore the continuing importance of terminology once a non-binary identity is established. Methods: This study uses thematic analysis on data produced from interviews with 16 participants who self-selected for the study and were recruited from several transgender and LGBTQ+ organisations on the basis that they identified outside the gender binary of male and female. Results: The analysis uncovered several key themes and sub-themes relating to terminology choice, encountering new terms and the process of identifying with new terminology, as well as becoming visible and understood by others. Conclusions: This study found that terminology is not only central in coming to identify as something other than exclusively male and female, it also remains an important factor when it comes to making a non-binary identity visible to others

Human mobility variations in response to restriction policies during the COVID-19 pandemic: An analysis from the Virus Watch community cohort in England, UK

Objective: Since the outbreak of COVID-19, public health and social measures to contain its transmission (e.g., social distancing and lockdowns) have dramatically changed people's lives in rural and urban areas globally. To facilitate future management of the pandemic, it is important to understand how different socio-demographic groups adhere to such demands. This study aims to evaluate the influences of restriction policies on human mobility variations associated with socio-demographic groups in England, UK. Methods: Using mobile phone global positioning system (GPS) trajectory data, we measured variations in human mobility across socio-demographic groups during different restriction periods from Oct 14, 2020 to Sep 15, 2021. The six restriction periods which varied in degree of mobility restriction policies, denoted as "Three-tier Restriction," "Second National Lockdown," "Four-tier Restriction," "Third National Lockdown," "Steps out of Lockdown," and "Post-restriction," respectively. Individual human mobility was measured with respect to the time period people stayed at home, visited places outside the home, and traveled long distances. We compared these indicators across the six restriction periods and across socio-demographic groups. Results: All human mobility indicators significantly differed across the six restriction periods, and the influences of restriction policies on individual mobility behaviors are correlated with socio-demographic groups. In particular, influences relating to mobility behaviors are stronger in younger and low-income groups in the second and third national lockdowns. Conclusions: This study enhances our understanding of the influences of COVID-19 pandemic restriction policies on human mobility behaviors within different social groups in England. The findings can be usefully extended to support policy-making by investigating human mobility and differences in policy effects across not only age and income groups, but also across geographical regions