Combining Double Fluorescence In Situ Hybridization with Immunolabelling for Detection of the Expression of Three Genes in Mouse Brain Sections

Detection of gene expression in different types of brain cells e.g., neurons, astrocytes, oligodendrocytes, oligodendrocyte precursors and microglia, can be hampered by the lack of specific primary or secondary antibodies for immunostaining. Here we describe a protocol to detect the expression of three different genes in the same brain section using double fluorescence in situ hybridization with two gene-specific probes followed by immunostaining with an antibody of high specificity directed against the protein encoded by a third gene. The Aspartoacyclase (ASPA) gene, mutations of which can lead to a rare human white matter disease - Canavan disease - is thought to be expressed in oligodendrocytes and microglia but not in astrocytes and neurons. However, the precise expression pattern of ASPA in the brain has yet to be established. This protocol has allowed us to determine that ASPA is expressed in a subset of mature oligodendrocytes and it can be generally applied to a wide range of gene expression pattern studies

The impact of public involvement in health research: what are we measuring? Why are we measuring it? Should we stop measuring it?

As public involvement in the design, conduct and dissemination of health research has become an expected norm and firmly enshrined in policy, interest in measuring its impact has also grown. Despite a drive to assess the impact of public involvement, and a growing body of studies attempting to do just this, a number of questions have been largely ignored. This commentary addresses these omissions: What is the impact of all this focus on measuring impact? How is the language of impact shaping the debate about, and the practice of, public involvement in health research? And how have shifting conceptualisations of public involvement in health research shaped, and been shaped by, the way we think about and measure impact? We argue that the focus on impact risks distorting how public involvement in health research is conceptualised and practised, blinding us to possible negative impacts. We call for a critical research agenda for public involvement that [a] considers public involvement not as an instrumental intervention but a social practice of dialogue and learning between researchers and the public; [b] explores how power relations play out in the context of public involvement in health research, what empowerment means and whose interests are served by it, and [c] asks questions about possible harms as well as benefits of public involvement, and whether the language of impact is helpful or not

Negotiating networks of self-employed work: strategies of minority ethnic contractors

Within the increased flexible, contracted work in cities, employment is negotiated through network arrangements characterised by multiplicity, mobility and fluidity. For black and minority ethnic group members, this network labour becomes fraught as they negotiate both their own communities, which can be complex systems of conflicting networks, as well as non-BME networks which can be exclusionary. This discussion explores the networking experiences of BME individuals who are self-employed in portfolio work arrangements in Canada. The analysis draws from a theoretical frame of ‘racialisation’ (Mirchandani and Chan, 2007) to examine the social processes of continually constructing and positioning the Other as well as the self through representations in these networks. These positions and concomitant identities enroll BME workers in particular modes of social production, which order their roles and movement in the changing dynamics of material production in networked employment

SYSTEMATIC REVIEW Involving older people in health research

Abstract Background: it is a UK policy requirement to involve patients and the public in health research as active partners. Objective: we reviewed published reports of studies which involved older people in commissioning, prioritising, designing, conducting or disseminating research. Search strategy and selection criteria: systematic searches of databases (PubMed, SCI-EXPANDED, SSCI, A&HCI, ASSIA, Embase, CINAHL and Medline) for English language studies published between 1995 and 2005 which had involved older people as partners in the research process as opposed to research subjects. Articles were reviewed by two authors using a standardised matrix for data extraction. Results: thirty studies were included and classified according to the stage in the research process in which older people were involved. Barriers to involving older people were: cultural divisions, language barriers, research skills capacity, ill health, time and resources. Four of the studies had been formally evaluated to identify the impact of involvement. Evaluation focussed on the impact on participants rather than on impact on research processes and outcomes. Benefits to participants included: increased knowledge, awareness and confidence, meeting others in similar situations, empowering older people to become active in their community regarding decisions/policies which affect them. Conclusions: factors hindering the involvement of older people in research were the same as reported factors hindering involvement of younger people, suggesting that age, per se, is not a barrier. To demonstrate the impact of user involvement on research quality, the definition of user involvement requires clarification, and systematic evaluation of research involving older people needs to be developed

Relation of Soil Acidity to Cotton Root Rot.

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Sucrose activates human taste pathways differently from artificial sweetener

Animal models suggest that sucrose activates taste afferents differently than non-caloric sweeteners. Little information exists how artificial sweeteners engage central taste pathways in the human brain. We assessed sucrose and sucralose taste pleasantness across a concentration gradient in 12 healthy control women and applied 10% sucrose and matched sucralose during functional magnet resonance imaging. The results indicate that (1) both sucrose and sucralose activate functionally connected primary taste pathways; (2) taste pleasantness predicts left insula response; (3) sucrose elicits a stronger brain response in the anterior insula, frontal operculum, striatum and anterior cingulate, compared to sucralose; (4) only sucrose, but not sucralose, stimulation engages dopaminergic midbrain areas in relation to the behavioral pleasantness response. Thus, brain response distinguishes the caloric from the non-caloric sweetener, although the conscious mind could not. This could have important implications on how effective artificial sweeteners are in their ability to substitute sugar intake

An 83 000-year-old ice core from Roosevelt Island, Ross Sea, Antarctica

In 2013 an ice core was recovered from Roosevelt Island, an ice dome between two submarine troughs carved by paleo-ice-streams in the Ross Sea, Antarctica. The ice core is part of the Roosevelt Island Climate Evolution (RICE) project and provides new information about the past configuration of the West Antarctic Ice Sheet (WAIS) and its retreat during the last deglaciation. In this work we present the RICE17 chronology, which establishes the depth–age relationship for the top 754 m of the 763 m core. RICE17 is a composite chronology combining annual layer interpretations for 0–343 m (Winstrup et al., 2019) with new estimates for gas and ice ages based on synchronization of CH4 and δ18Oatm records to corresponding records from the WAIS Divide ice core and by modeling of the gas age–ice age difference. Novel aspects of this work include the following: (1) an automated algorithm for multiproxy stratigraphic synchronization of high-resolution gas records; (2) synchronization using centennial-scale variations in methane for pre-anthropogenic time periods (60–720 m, 1971 CE to 30 ka), a strategy applicable for future ice cores; and (3) the observation of a continuous climate record back to ∼65 ka providing evidence that the Roosevelt Island Ice Dome was a constant feature throughout the last glacial period