“Who am I?” How Female Care Leavers Construct and Make Sense of Their Identity

Identity formation may be more complex for those who have been in foster care in the face of childhood abuse, difficult relationships, unstable environments, and multiple care contexts but this does not imply there is anything pathological about it. Given the higher levels of mental health difficulties in looked after children and the known role identity has in mental health, whether as a risk or a protective factor, it seems clinically significant to investigate what factors help construct or hinder the formation of identity for those who have been in care. Interpretative Phenomenological Analysis was used to analyze semistructured interviews of eight female care leavers about the understanding of their identity development. Three superordinate themes emerged which encapsulated participants’ identity development. These included Construction of Identity—How I Became Me, Understanding of Identity—Who am I, and Experience of Identity—How My Identity Plays Out. Participants’ construction of identity can be understood in the context of early adverse environments and developmental trauma. This construction of self, in turn, mediates how participants understand and experience their identity. Findings were discussed in relation to previous research, and limitations were outlined. Implications for future research included giving fuller consideration to the role of developmental trauma in identity formation. Clinical implications encourage understanding of looked after children and care leavers in the context of developmental trauma, rather than focusing on symptoms of various diagnoses

Native Ambient Mass Spectrometry Enables Analysis of Intact Endogenous Protein Assemblies up to 145 kDa Directly from Tissue

[Image: see text] Untargeted label-free interrogation of proteins in their functional form directly from their physiological environment promises to transform life sciences research by providing unprecedented insight into their transient interactions with other biomolecules and xenobiotics. Native ambient mass spectrometry (NAMS) shows great potential for the structural analysis of endogenous protein assemblies directly from tissues; however, to date, this has been limited to assemblies of low molecular weight (<20 kDa) or very high abundance (hemoglobin tetramer in blood vessels, RidA homotrimer in kidney cortex tissues). The present work constitutes a step change for NAMS of protein assemblies: we demonstrate the detection and identification of a range of intact endogenous protein assemblies with various stoichiometries (dimer, trimer, and tetramer) from a range of tissue types (brain, kidney, liver) by the use of multiple NAMS techniques. Crucially, we demonstrate a greater than twofold increase in accessible molecular weight (up to 145 kDa). In addition, spatial distributions of protein assemblies up to 94 kDa were mapped in brain and kidney by nanospray desorption electrospray ionization (nano-DESI) mass spectrometry imaging

Laser capture microdissection and native mass spectrometry for spatially-resolved analysis of intact protein assemblies in tissue †

Previously, we have shown that native ambient mass spectrometry imaging allows the spatial mapping of folded proteins and their complexes in thin tissue sections. Subsequent top-down native ambient mass spectrometry of adjacent tissue section enables protein identification. The challenges associated with protein identification by this approach are (i) the low abundance of proteins in tissue and associated long data acquisition timescales and (ii) irregular spatial distributions which hamper targeted sampling of the relevant tissue location. Here, we demonstrate that these challenges may be overcome through integration of laser capture microdissection in the workflow. We show identification of intact protein assemblies in rat liver tissue and apply the approach to identification of proteins in the granular layer of rat cerebellum

Tissue Washing Improves Native Ambient Mass Spectrometry Detection of Membrane Proteins Directly from Tissue

Native ambient mass spectrometry enables the in situ analysis of proteins and their complexes directly from tissue, providing both structural and spatial information. Until recently, the approach was applied exclusively to the analysis of soluble proteins; however, there is a drive for new techniques that enable analysis of membrane proteins. Here we demonstrate native ambient mass spectrometry of membrane proteins, including β-barrel and α-helical (single and multipass) integral membrane proteins and membrane-associated proteins incorporating lipid anchors, by integration of a simple washing protocol to remove soluble proteins. Mass spectrometry imaging revealed that washing did not disrupt the spatial distributions of the membrane and membrane-associated proteins. Some delocalization of the remaining soluble proteins was observed

Quantitative Characterization of Three Carbonic Anhydrase Inhibitors by LESA Mass Spectrometry

[Image: see text] Liquid extraction surface analysis (LESA) coupled to native mass spectrometry (MS) presents unique analytical opportunities due to its sensitivity, speed, and automation. Here, we examine whether this tool can be used to quantitatively probe protein–ligand interactions through calculation of equilibrium dissociation constants (K(d) values). We performed native LESA MS analyses for a well-characterized system comprising bovine carbonic anhydrase II and the ligands chlorothiazide, dansylamide, and sulfanilamide, and compared the results with those obtained from direct infusion mass spectrometry and surface plasmon resonance measurements. Two LESA approaches were considered: In one approach, the protein and ligand were premixed in solution before being deposited and dried onto a solid substrate for LESA sampling, and in the second, the protein alone was dried onto the substrate and the ligand was included in the LESA sampling solvent. Good agreement was found between the K(d) values derived from direct infusion MS and LESA MS when the protein and ligand were premixed; however, K(d) values determined from LESA MS measurements where the ligand was in the sampling solvent were inconsistent. Our results suggest that LESA MS is a suitable tool for quantitative analysis of protein–ligand interactions when the dried sample comprises both protein and ligand