Involving children, teachers and parents/carers in dialogues around child well-being in schools

Building dynamic partnerships between communities and academic institutions, via knowledge exchange, plays a vital role in generating research with mutual benefits for research partners and wider communities. Knowledge exchange between universities and schools has recently received particular interest in child and adolescent mental health research. This article outlines a knowledge exchange programme that involved children, primarily, and teachers and parents/carers in dialogues around school well-being. Using a series of co-created and multimodal creative activities, we: (1) worked closely with 25 students of an inclusive and multicultural primary school (aged 9 to 10 years old) to understand their school well-being experiences, and to co-create a school blog to communicate key messages with local communities; and (2) initiated a discussion around child and school well-being with a group of parents/carers and teachers through expanding on the children’s blog. In this article, we outline our methodology to facilitate children’s involvement with the project well-being activities. We explore key methodological strengths and challenges, and highlight lessons we learned and how these stress the significance of seeking young people’s points of view when designing school well-being initiatives

The impact of active HIV-1 replication on the physiological age-related decline of immature-transitional B-cells in HIV-1 infected children

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Comparison of thermal effects of stilbenoid analogs in lipid bilayers using differential scanning calorimetry and molecular dynamics: correlation of thermal effects and topographical position with antioxidant activity

In previous studies it was shown that cannabinoids (CBs) bearing a phenolic hydroxyl group modify the thermal properties of lipid bilayers more significantly than methylated congeners. These distinct differential properties were attributed to the fact that phenolic hydroxyl groups constitute an anchoring group in the vicinity of the headgroup, while the methylated analogs are embedded deeper towards the hydrophobic region of the lipid bilayers. In this work the thermal effects of synthetic polyphenolic stilbenoid analogs and their methylated congeners have been studied using differential scanning calorimetry (DSC).Molecular dynamics (MD) simulations have been performed to explain the DSC results. Thus, two of their phenolic hydroxyl groups orient in the lipid bilayers in such a way that they anchor in the region of the headgroup. In contrast, their methoxy congeners cannot anchor effectively and are embedded deeper in the hydrophobic segment of the lipid bilayers. The MD results explain the fact that hydroxystilbenoid analogs exert more significant effects on the pretransition than their methoxy congeners, especially at low concentrations. To maximize the polar interactions, the two phenolic hydroxyl groups are localized in the vicinity of the head-group region, directing the remaining hydroxy group in the hydrophobic region. This topographical position of stilbenoid analogs forms a mismatch that explains the significant broadening of the width of the phase transition and lowering of the main phasetransition temperature in the lipid bilayers. At high concentrations, hydroxy and nonhydroxy analogs appear to form different domains. The correlation of thermal effects with antioxidant activity is discusse

Intradermal Electroporation of Naked Replicon RNA Elicits Strong Immune Responses

RNA-based vaccines represent an interesting immunization modality, but suffer from poor stability and a lack of efficient and clinically feasible delivery technologies. This study evaluates the immunogenic potential of naked in vitro transcribed Semliki Forest virus replicon RNA (RREP) delivered intradermally in combination with electroporation. Replicon-immunized mice showed a strong cellular and humoral response, contrary to mice immunized with regular mRNA. RREP-elicited induction of interferon-γ secreting CD8+ T cells and antibody responses were significantly increased by electroporation. CD8+ T cell responses remained substantial five weeks post vaccination, and antigen-specific CD8+ T cells with phenotypic characteristics of both effector and central memory cells were identified. The immune response during the contraction phase was further increased by a booster immunization, and the proportion of effector memory cells increased significantly. These results demonstrate that naked RREP delivered via intradermal electroporation constitute an immunogenic, safe and attractive alternative immunization strategy to DNA-based vaccines

T cells and costimulatory factors in myasthenia gravis

The autoimmune disease myasthenia gravis (MG) is characterized by muscle weakness due to a loss of acetylcholine receptors (AChR) at the neuromuscular end plate. Most MG patients have pathogenic antibodies directed against the receptor. We provide further evidence that T cells are important for establishment and continuation of the disease. We have detected pathogenic antibodies capable of transferring disease to mice in healthy twin sisters as well as in their MG affected sisters in two monozygotic twin pairs discordant for MG. However, the healthy twin sisters did not demonstrate T cell responses against the AChR. A further support to the importance of T cells in maintaining the disease is shown in a successful treatment of an MG patient with antibodies targeting CD25. This molecule is expressed on activated cells. The levels of activated T cells, serum levels of IL-10 as well as the soluble costimulatory molecules sCD28, sCD80, sCD86 and sCD152 decreased, suggesting a normalization of an abnormally activated immune system. Costimulatory molecules are important in the activation and inhibition of an immune response. We demonstrated reduced expression of the costimulatory molecule CD152 (cytotoxic T lymphocyte associated antigen 4, CTLA-4) in T cells from MG patients. CD152 is essential to inhibit an immune response, therefore the patients might have a reduced potential to down-regulate an ongoing immune reaction. We observed that the G allele at position +49 in coding sequence 1 of the CD152 gene was associated to increased immune activity, manifested as increased levels of IL-1beta and CD3+CD28+ cells. MG patients with thymoma more frequently had the G/G genotype or the G allele, which could explain a more active immune response in patients with this genotype. The costimulatory factors CD28, CD80, CD86 and CD152 also exist in soluble forms. The concentrations of sCD28, sCD80, sCD86 and sCD152, all of which recently have been shown to be increased in different diseases, were not increased in MG patients. However, in one of our studies we detected elevated levels of sCD152 in MG patients. The concentrations of sCD28, sCD80, sCD86 and sCD152 correlated to each other and to IL-6, IL-10 and IFN-gamma. All four soluble costimulatory molecules correlated to sCD25 in healthy persons, while only sCD80 and sCD86 correlated to sCD25 in MG patients. In addition, we confirmed results by others demonstrating increased serum levels of sCD25 and sICAM-1. We produced a recombinant form of the naturally occurring soluble costimulatory factor CD80. Recombinant sCD80 demonstrated capacity to interact with its natural ligands CD28 and CD152. It preferentially bound to activated cells. In addition, it displayed immunosuppressive properties, demonstrated by inhibition of T cell activation, inhibition of the mixed lymphocyte reaction and the ability to alter the cytokine secretion balance in vitro. The effect of sCD80 in vivo has to be clarified, but it is tempting to speculate about a potential future use of the soluble protein in treatment of diseases like MG. In summary, we have provided further evidence that T cells are important in the initiation and maintenance of MG, and that the costimulatory factors could be involved in disease progression

Cellular immune response 5 weeks post immunization.

<p>(a) Antigen-specific IFN-γ positive CD8⁺ T cells per million splenocytes, (b) Proportion responding CD8⁺ T cells as determined by intracellular staining of IFN-γ, IL-2 and TNF after SIINFEKL-peptide stimulation, or (c) proportion effector memory CD8⁺ T cells (pentamer H-2Kb/SIINFEKL positive CD8⁺CD62L⁻CD127⁺ cells) 5 weeks after the last intradermal immunization in combination with electroporation. Mice were either given one immunization (1×5 µg) or two immunizations 5 weeks apart (2×5 µg). Data shows average number of positive cells with error bars showing standard error of the mean. A booster immunization significantly increased the cellular memory response for both RREP-tLuc and DREP-tLuc (p<0.01). The total number of mice per group is indicated in parenthesis above each bar.</p

Immune responses after electroporation.

<p>(a) Antigen-specific IFN-γ positive CD8⁺ T cells per million splenocytes ten days after intradermal immunization either with or without electroporation. Electroporation significantly increases the number of positive cells in RREP-tLuc and DREP-tLuc immunized animals (p<0.05 and p<0.001 respectively). Data shows average number of positive cells from two separate experiments with error bars showing standard error of the mean. The total number of mice analyzed is indicated in parenthesis above each bar. (b) ELISA for total anti-β-galactosidase IgG antibody responses in sera from immunized mice 14 days post immunization. Electroporation significantly increases the antibody responses for both RREP-β-gal and DREP-β-gal (p<0.001). Data are plotted as reciprocal end-point titers and shown for each individual mouse. The total number of mice analyzed is indicated in parenthesis and the percentage of responding mice is indicated above each group. <i>Abbreviations</i>: SFC = spot forming cells, E.P. = electroporation, and Ab = antibody.</p

Comparison of thermal effects of stilbenoid analogs in lipid bilayers using differential scanning calorimetry and molecular dynamics: correlation of thermal effects and topographical position with antioxidant activity

In previous studies it was shown that cannabinoids (CBs) bearing a phenolic hydroxyl group modify the thermal properties of lipid bilayers more significantly than methylated congeners. These distinct differential properties were attributed to the fact that phenolic hydroxyl groups constitute an anchoring group in the vicinity of the head-group, while the methylated analogs are embedded deeper towards the hydrophobic region of the lipid bilayers. In this work the thermal effects of synthetic polyphenolic stilbenoid analogs and their methylated congeners have been studied using differential scanning calorimetry (DSC). Molecular dynamics (MD) simulations have been performed to explain the DSC results. Thus, two of their phenolic hydroxyl groups orient in the lipid bilayers in such a way that they anchor in the region of the head-group. In contrast, their methoxy congeners cannot anchor effectively and are embedded deeper in the hydrophobic segment of the lipid bilayers. The MD results explain the fact that hydroxystilbenoid analogs exert more significant effects on the pretransition than their methoxy congeners, especially at low concentrations. To maximize the polar interactions, the two phenolic hydroxyl groups are localized in the vicinity of the head-group region, directing the remaining hydroxy group in the hydrophobic region. This topographical position of stilbenoid analogs forms a mismatch that explains the significant broadening of the width of the phase transition and lowering of the main phase-transition temperature in the lipid bilayers. At high concentrations, hydroxy and nonhydroxy analogs appear to form different domains. The correlation of thermal effects with antioxidant activity is discussed