Piloting Lead with Love: a film-based intervention to improve parents' responses to their lesbian, gay, and bisexual children.

Lesbian, gay, and bisexual youth are at increased risk for a variety of poor health outcomes, relative to their heterosexual counterparts, and recent research implicates family responses to a child's sexual orientation as an important predictor of these health difficulties. Lead with Love is a 35-min documentary-style preventive intervention created to improve parents' behaviors toward their lesbian, gay, and bisexual (LGB) children, by providing parents with support, information, and concrete behavioral guidance. The film was made available free online, and was promoted widely with a multi-media marketing campaign. In this paper we describe the theoretical and empirical rationale for the intervention, and report findings from pilot data collected in the first year after the film's release. Specifically, we gathered data to examine the feasibility of reaching parents of LGB youth with this intervention, to determine whether it was acceptable, and to provide preliminary indicators of its potential efficacy. In the first 12 months after launch, 10,949 individuals viewed the film online. The film successfully reached parents of LGB youth (n=1,865), including the hardest to reach parents: 21% had only learned about their child's sexual orientation in the past month, 36% reported having an LGB child was "very" or "extremely" hard for them, and 86% had never obtained any other formal support for having an LGB child. Parents who completed a follow-up assessment immediately after the film reported significant pre- to post-film increases in self-efficacy for parenting an LGB child.

Structure of the Fab fragment of the anti-murine EGFR antibody 7A7 and exploration of its receptor binding site.

The EGF receptor is an important target of cancer immunotherapies. The 7A7 monoclonal antibody has been raised against the murine EGFR, but it cross-reacts with the human receptor. The results from experiments using immune-competent mice can therefore, in principle, be extrapolated to the corresponding scenario in humans. In this work we report the crystal structure of the 7A7 Fab at an effective resolution of 1.4Å. The antibody binding site comprises a deep pocket, located at the interface between the light and heavy chains, with major contributions from CDR loops H1, H2, H3 and L1. Binding experiments show that 7A7 recognizes a site on the EGFR extracellular domain that is not accessible in its most stable conformations, but that becomes exposed upon treatment with a tyrosine kinase inhibitor. This suggests a recognition mechanism similar to that proposed for mAb 806.

Function of the CysD domain of the gel-forming MUC2 mucin.

The colonic human MUC2 mucin forms a polymeric gel by covalent disulfide bonds in its N- and C-termini. The middle part of MUC2 is largely composed of two highly O-glycosylated mucin domains that are interrupted by a CysD domain of unknown function. We studied its function as recombinant proteins fused to a removable immunoglobulin Fc domain. Analysis of affinity-purified fusion proteins by native gel electrophoresis and gel filtration showed that they formed oligomeric complexes. Analysis of the individual isolated CysD parts showed that they formed dimers both when flanked by two MUC2 tandem repeats and without these. Cleavages of the two non-reduced CysD fusion proteins and analysis by MS revealed the localization of all five CysD disulfide bonds and that the predicted C-mannosylated site was not glycosylated. All disulfide bonds were within individual peptides showing that the domain was stabilized by intramolecular disulfide bonds and that CysD dimers were of non-covalent nature. These observations suggest that CysD domains act as non-covalent cross-links in the MUC2 gel, thereby determining the pore sizes of the mucus.

Preparing staff to provide bereavement support.

An in-service training programme was developed to meet the needs of an NHS palliative care service for children and young people, specifically focused on bereavement work. A training day was held every six months that allowed staff to share and support each other's practice and to consider how best practice could be promoted. Each training day was evaluated by attendees, who were asked to rate the usefulness of the day and give qualitative feedback. After four training days had been completed, staff views on the impact and benefits of the programme were sought using a questionnaire that incorporated a set of statements highlighting factors important to good bereavement care. Participant evaluations indicated that the aims of the training days were being met. Responses to the questionnaire were positive, both in terms of improvement and positive outcomes in the workplace. Ongoing training and support as well as clinical supervision are essential to maintain good practice in palliative and bereavement care.

Use of and satisfaction with Community Aged Care Packages in the eastern suburbs of Sydney.

OBJECTIVE: The pilot study aimed to examine the accessibility and the flexibility of the Community Aged Care Package (CACP) program, and provide recommendations for further improvement. METHOD: Data were collected using structured interviews with 80 CACP recipients, and mail surveys with nine service coordinators of CACP services. Descriptive statistics and χ2 analysis were used for quantitative information, and thematic analysis for qualitative data. RESULTS: CACPs were utilised more frequently and for longer periods by clients with English-speaking backgrounds and those living alone. The average level of client satisfaction with CACP was high, over 62% rating extremely satisfied. Participants expressed concerns related to CACPs including inflexibility, lack of communication between service providers and other health services, poor continuity and quality of care, inadequate funding, problems with recruitment, retention and support for staff. CONCLUSION: While the clients' overall satisfaction levels were rated high, qualitative information suggested a need for improvement of the current delivery of the CACP program.

Controlling electron transfer in Acyl-CoA oxidases and dehydrogenases: a structural view.

Plants produce a unique peroxisomal short chain-specific acyl-CoA oxidase (ACX4) for beta-oxidation of lipids. The short chain-specific oxidase has little resemblance to other peroxisomal acyl-CoA oxidases but has an approximately 30% sequence identity to mitochondrial acyl-CoA dehydrogenases. Two biochemical features have been linked to structural properties by comparing the structures of short chain-specific Arabidopsis thaliana ACX4 with and without a substrate analogue bound in the active site to known acyl-CoA oxidases and dehydrogenase structures: (i) a solvent-accessible acyl binding pocket is not required for oxygen reactivity, and (ii) the oligomeric state plays a role in substrate pocket architecture but is not linked to oxygen reactivity. The structures indicate that the acyl-CoA oxidases may encapsulate the electrons for transfer to molecular oxygen by blocking the dehydrogenase substrate interaction site with structural extensions. A small binding pocket observed adjoining the flavin adenine dinucleotide N5 and C4a atoms could increase the number of productive encounters between flavin adenine dinucleotide and O2.

The affinity of the FimH fimbrial adhesin is receptor-driven and quasi-independent of Escherichia coli pathotypes.

Type-1 fimbriae are important virulence factors for the establishment of Escherichia coli urinary tract infections. Bacterial adhesion to the high-mannosylated uroplakin Ia glycoprotein receptors of bladder epithelium is mediated by the FimH adhesin. Previous studies have attributed differences in mannose-sensitive adhesion phenotypes between faecal and uropathogenic E. coli to sequence variation in the FimH receptor-binding domain. We find that FimH variants from uropathogenic, faecal and enterohaemorrhagic isolates express the same specificities and affinities for high-mannose structures. The only exceptions are FimHs from O157 strains that carry a mutation (Asn135Lys) in the mannose-binding pocket that abolishes all binding. A high-mannose microarray shows that all substructures are bound by FimH and that the largest oligomannose is not necessarily the best binder. Affinity measurements demonstrate a strong preference towards oligomannosides exposing Manalpha1-3Man at their non-reducing end. Binding is further enhanced by the beta1-4-linkage to GlcNAc, where binding is 100-fold better than that of alpha-d-mannose. Manalpha1-3Manbeta1-4GlcNAc, a major oligosaccharide present in the urine of alpha-mannosidosis patients, thus constitutes a well-defined FimH epitope. Differences in affinities for high-mannose structures are at least 10-fold larger than differences in numbers of adherent bacteria between faecal and uropathogenic strains. Our results imply that the carbohydrate expression profile of targeted host tissues and of natural inhibitors in urine, such as Tamm-Horsfall protein, are stronger determinants of adhesion than FimH variation.