Search

Suicide is a major public health concern and leading cause of death among men in Canada. This study reports the feasibility and acceptability of Buddy Up, a peer-based suicide prevention campaign for men. A mixed-methods approach was used to analyze respondent survey questionnaires (n= 48) and individual participant interviews (n= 19) collected from campaign users. Survey respondents reported that they enjoyed their involvement in the campaign (92%), were more confident to talk with men about mental health and suicide (95%), and would recommend Buddy Up to others (95%). Qualitative interviews were thematically analyzed to develop three inductively derived themes: (a) Engaging men with relatable masculine content and design: “Buddy Up really spoke to them in their language,” highlighting the importance of understanding and working with gendered practices and motivations to legitimize and motivate involvement in suicide prevention; (b) Leveraging campaign participation to initiate conversations and promote mental health: “It gives men language and license to start asking questions,” revealing ways in which participants utilized Buddy Up to negotiate and norm checking-in to promote men’s mental health; and (c) Driving new masculine cultures: “We start every meeting with a mental health moment,” identifying how participants fostered healthy milieus for disclosing mental health challenges with teamwork and preventive action under the banner of Buddy Up. The study findings support the feasibility of Buddy Up and highlight the acceptability of peer-based approaches to mental health promotion. The findings can also empirically guide future efforts for systematically building men’s peer-based suicide prevention programs.

Queer men (i.e., men who are not heterosexual; sexual minority men) are disproportionately affected by negative body image - they experience greater body dissatisfaction are more likely to develop eating disorders than heterosexual men. While existing literature has examined individual-level predictors of negative body image for queer men, less is known about why queer men as a group are disproportionately affected by negative body image. By synthesising existing theoretical frameworks, research, policy, and media reporting, this narrative review moves towards an understanding of systemic-level negative body image for queer men. Through the lens of hegemonic masculinity, we explain how systemic experiences of stigma work to inform unattainable appearance standards for queer men, and how these standards then contribute to pervasive negative body image concerns among this community. Next, we describe how systemic stigma works to exacerbate negative health outcomes for queer men with body image concerns. Finally, we present a synthesized model of the processes outlined in this review, articulate testable predictions for future studies, and describe practical implications that could be widely employed to improve body image for queer men. Our review is the first to propose a comprehensive explanation of systemic negative body image for queer men.

Background: Helplines are an accessible form of support for people struggling with difficulties in their lives and are key services in suicide prevention and intervention. Men's experiences of telephone helplines are not well understood, despite high male suicide rates. Methods: We conducted an online cross-sectional survey with N = 684 Australian men (aged 17-83 years, M = 50.13) using open- and closed-ended questions about their experiences of helplines during the COVID-19 pandemic. Descriptive statistics were analysed to investigate differences between men using and not using helplines. Qualitative responses were analysed using thematic analysis. Results: Within the sample, 100 men (14.6%) had used a helpline service. Men using helplines were more likely to be unemployed and in younger age brackets than those not using helplines. They were also more likely to report experiencing stressors related to COVID-19, including financial stress and job loss, perceived impact on mental health and relationship breakdown. Qualitative analysis indicated varied experiences of helplines, with men shedding light on how their interaction with a counsellor, the structure of services and their expectations of the service impacted their experience. Conclusions: Further in-depth qualitative enquiry in this space is required, with the objective of understanding how helpline services may seek to better engage with male callers.

Despite known links between men's avoidant coping behaviours (e.g., distraction, denial, substance use) and suicide risk, little research has explored the mechanisms underpinning this relationship. This study sought to examine whether male-type depression symptoms (e.g., anger, aggression, emotion suppression), assessed by the Male Depression Risk Scale, mediate the association between avoidant coping and suicide/self-harm ideation in men. Data were drawn from an online survey of a community sample of 606 Australian men (M age = 50.11 years; SD = 15.00), conducted during the COVID-19 pandemic. Mediation analyses were applied to examine the effect of male-type depression on the association between avoidant coping and suicidal/self-harm ideation, controlling for age, resilience and the experience of two psychosocial stressors during the COVID-19 pandemic (financial stress and government restrictions). Avoidant coping was associated with suicidal/self-harm ideation, r = 0.45, p < 0.001. Results supported a mediating role of male-type depression symptoms in this relationship, R2= 0.29, PM = 0.36, p < 0.001, underscoring the importance of screening for male-type depression symptoms to better identify men at risk of suicidal/self-harm ideation. Results also suggest a need to support men to develop effective coping strategies, particularly in the context of common psychosocial stressors experienced during the COVID-19 pandemic and beyond.

The COVID-19 pandemic has presented a suite of circumstances that will simultaneously affect mental health and mobilize coping strategies in response. Building on a lack of research specifically exploring men's mental health impacts during the COVID-19 pandemic, this study presents the results of a qualitative survey exploring men's self-reported aspects of the pandemic giving rise to mental health challenges, alongside their diverse coping strategies applied during this time. The sample comprised 555 men from North America (age M = 38.8 years; SD = 13.5 years), who participated via an online survey with two open-ended qualitative questions assessing, respectively, the aspects of the pandemic affecting their mental health, and the strategies used to manage these challenges. Free-text responses were coded using inductive content analysis. Results pertaining to the mental health impacts of COVID-19 were categorized into two overarching themes: far-reaching ramifications of COVID-19 encompassing consequences for lifestyle, work, and functioning, alongside novel anxieties related to health risks and daily uncertainty. In addition, coping strategies reported were categorized into two broad themes: efforts to avoid, dull or distract oneself from distress, alongside adapting and doing things differently, which encompassed largely approach-oriented efforts to flexibly ameliorate distress. Results signal the far-reaching impacts of COVID-19, alongside profound flexibility and diverse enactments of resilience among men in adapting to unprecedented challenges. Findings have implications for mental health promotion that should aim to leverage men's adaptive coping to encourage opportunities for social connectedness in response to the mental health impacts of the various psychosocial challenges of the COVID-19 pandemic.

The capacity for boys' and young men's mental health promotion to act via shifting masculine norms that are linked to poor mental health outcomes, highlights the need to improve the extent to which school-based programs can promote mental health through leveraging more positive embodiments of masculinity. To-date, the perspectives of parents and teachers on such processes are understudied. This qualitative study presents teacher and parent views regarding adolescent masculinities and avenues for school-based developmental programming for boys and young men. In this study, 16 individual qualitative interviews were undertaken with 10 parents (six females, four males), and six teachers (three females, three males), recruited from an independent all-boys' grammar school in Melbourne, Australia. Thematic analysis of parents' and teachers' perspectives indicated their perception of the role of context-dependent "public" and "private" masculinities, the influence of Australian masculinity norms, and the role of private boys' school cultures in the development of adolescent masculinities. Additionally, strategies for development encompassed participants' appetite for boys' exposure to positive role models, in addition to consistent and relevant developmental programming to support positive masculinity development. Findings have implications for efforts to support prosocial masculine identity development via school-based initiatives, as an avenue to promoting mental health of boys and young men.

BACKGROUND: Although the proportion of men seeking professional mental health care has risen over the past two decades, on average, men continue to attend fewer sessions of psychotherapy and are more likely to drop out of treatment prematurely compared to women. Men account for three-quarters of suicide deaths; furthermore, over half of the males who die by suicide have engaged with mental health care in the 12 months prior to their death. These findings highlight a need to equip mental health practitioners with skills to improve male clients' engagement and mental health outcomes. This article reports the protocol for a randomized controlled trial of Men in Mind, a self-paced online training program purpose-built to advance the clinical competencies of practitioners who provide psychotherapy to male clients. METHODS: A randomized controlled trial with two parallel groups will be conducted. Participating practitioners will be randomly allocated, on a 1:1 basis, to the intervention group (Men in Mind training) or a waitlist control group. The primary outcome, efficacy of the training, will be evaluated by pre- to post-training (T1 to T2) changes in scores on the Engaging Men in Therapy Scale (EMITS) in the intervention group, relative to the control group. DISCUSSION: This trial will provide evidence of the efficacy of Men in Mind training, as an interim step towards adjusting content and delivery of the intervention to maximize the potential for sustaining and scaling. TRIAL REGISTRATION: The trial was registered prospectively with the Australian New Zealand Clinical Trials Registry on 3rd December 2021 (ACTRN12621001669886).

The COVID-19 pandemic restrictions, uncertainties and management inconsistencies have been implicated in men's rising distress levels, which in turn have somewhat normed the uptake of telemental healthcare services (i.e., phone and/or video-conference-based therapy). Given past evidence of poor engagement with telemental health among men, this mixed-methods study examined Australian men's use of, and experiences with telemental health services relative to face-to-face care during the pandemic. A community sample of Australian-based men (N = 387; age M = 47.5 years, SD = 15.0 years) were recruited via Facebook advertising, and completed an online survey comprising quantitative items and open-response qualitative questions with the aim of better understanding men's experiences with telemental healthcare services. In total, 62.3% (n = 241) of participants reported experience with telemental health, and regression analyses revealed those who engaged with telemental health were on average younger, more likely to be gay and university educated. Men who had used telemental health were, on average, more satisfied with their therapy experience than those who had face-to-face therapy. Among those who had telemental healthcare, marginally lower satisfaction was observed among regional/rural based relative to urban men, and those who had to wait longer than 2 months to commence therapy. Qualitative findings highlighted positive aspects of telemental healthcare including comfort with accessing therapy from familiar home environments and the convenience and accessibility of telemental health alongside competing commitments and COVID-19 restrictions. Conversely, drawbacks included technical limitations such as crosstalk impeding therapeutic progress, disconnects and audio-visual lag-times and the 'impersonal' nature of telemental healthcare services. Findings broadly signal COVID-19 induced shifts norming of the use of virtual therapy services, with clear scope for improvement in

BACKGROUND: We have recently reported on a recombinant von Willebrand factor (VWF) D'D3 albumin fusion protein (rD'D3-FP) developed to extend the half-life of coagulation factor VIII (FVIII) for the treatment of hemophilia A. Based on predictive modelling presented in this study, we hypothesized that modifying rD'D3-FP to improve FVIII interaction would reduce exchange with endogenous VWF and provide additional FVIII half-life benefit. OBJECTIVES: The aim of this study was to identify novel rD'D3-FP variants with enhanced therapeutic efficacy in extending FVIII half-life. METHODS: Through both directed mutagenesis and random mutagenesis using a novel mammalian display platform, we identified novel rD'D3-FP variants with increased affinity for FVIII (rVIII-SingleChain) under both neutral and acidic conditions and assessed their ability to extend FVIII half-life in vitro and in vivo. RESULTS: In rat preclinical studies, rD'D3-FP variants with increased affinity for FVIII displayed enhanced potency, with reduced dose levels required to achieve equivalent rVIII-SingleChain half-life extension. In cell-based imaging studies in vitro, we also demonstrated reduced dissociation of rVIII-SingleChain from the rD'D3-FP variants within acidic endosomes and more efficient co-recycling of the rD'D3-FP/rVIII-SingleChain complex via the FcRn recycling system. CONCLUSIONS: In summary, at potential clinical doses, the rD'D3-FP variants provide marked benefits with respect to dose levels and half-life extension of co-administered FVIII, supporting their development for use in the treatment of hemophilia A.

Introduction Current gendered health disparities impacting the wellbeing of boys and young men require new early intervention-focussed approaches. Health promotion programs developed with young men’s health needs and preferences in mind commonly report positive outcomes. Male-specific rite of passage programs aim to formally acknowledge the life-stage transition from boyhood to manhood through a holistic focus on identity, community, and social responsibility. While these programs are growing in popularity, there is limited data available on their effectiveness. Methods This study undertook a pilot evaluation of the Making of Men rite of passage program in a sample of secondary school boys (n=61, age M=16.0, SD=0.5) and their accompanying fathers or male mentors (n=47, age M=52.1, SD=5.8 years) providing non-matched pre-test, post-test, alongside follow-up data for participating boys. Qualitative interviews were also undertaken with 15 individuals (5 mothers, 6 staff members, 4 fathers). Results Quantitative program feedback indicated acceptability, with most respondents providing positive feedback, particularly from participating fathers. Exploratory quantitative effects indicated potential improvements in subjective social support and open communication among boys. Fathers appeared to report lower conformity to traditional masculine norms post-program, in addition to more open communication. Qualitative interviews identified three main themes: enabling relational bonds, creating a men-specific context, and supporting developmental transitions. Conclusions Positive program acceptability and promising outcome effects highlighted the present rite of passage program as a promising mechanism for supporting healthy masculine identity development among adolescent males. Finally, limitations and future directions are discussed, particularly in terms of gender equality integration for rite of passage programs.

While increasing numbers of Australian men are accessing mental health services, the sustainability of their therapy engagement varies significantly, with many men being lost to follow-up. The current study investigated dropout rates in a large community-based male sample to highlight the reasons for, and potential predictors of, men dropping out of mental health care services. Data were drawn from an online survey of 1907 Australian men (aged 16-85; M = 44.1 years) reflecting on their broad experiences in mental health therapy. Participants responded to bespoke items assessing their past dropout experience and reasons for dropping out, the odds of which were modeled in relation to demographics and predictors (e.g., therapist engagement strategies, alignment to traditional masculinity and pre-therapy feelings of optimism, shame, and emasculation). The overall dropout rate from therapy was 44.8% (n = 855), of which 26.6% (n = 120) accessed therapy once and did not return. The most common reasons for dropout were lack of connection with the therapist (54.9%) and the sense that therapy lacked progress (20.2%). Younger age, unemployment, self-reported identification with traditional masculinity, the presence of specific therapist engagement strategies, and whether therapy made participants feel emasculated all predicted dropout. Current depressive symptoms and suicidality were also higher amongst dropouts. Therapists should aim to have an honest discussion with all clients about the importance of therapy fit, including the real likelihood of dropout, in order to ensure this does not deter future engagement with professional services.

While research has explored the intersection between masculinities and men's experiences of suicidality, comparatively little attention has been paid to the stories surrounding suicidal men's decision to seek help. The ways in which men experiencing suicidal thoughts and behaviors embody masculinities alongside their enlistment of mental health services remains largely unknown. The present study explored 262 Australian men's stories surrounding the impetus for help-seeking for suicidal thoughts and behaviors. The sample comprised men ranging in age from 17 to 74 years (M = 40.99; SD = 15.92 years), with most participants residing in a metropolitan area (55.3%), employed full time (43.1%), non-indigenous (95.4%) and heterosexual (73.7%). Participants elaborated on their reasons for help-seeking via an open-text qualitative survey, delivered as part of a larger study exploring help-seeking experiences of Australian men. Thematic analysis of responses generated four themes highlighting the diversity of experience across men, with some highlighting impacts of emasculating early trauma(s) on their suicidality, while others reflected an impulsiveness tied to situational stressors that fractured their masculine identity (e.g., relationship breakdown; job loss). Many men had epiphanies as they reached the limits of their self-reliance and came to terms with their need for help. As their suicidality was witnessed by-and began to impact-those around them, the sight of their previously masked pain by others often facilitated their help-seeking journey. The present findings underscore the complex and multifactorial role of masculinities in men's suicidality and their paths to help-seeking. Important inroads for future public mental health promotion efforts are discussed, in terms of leveraging self-reliant and caring masculinities in helping men to develop healthy coping in the context of suicidality.

OBJECTIVE: Emerging research highlights that therapists experience difficulty engaging and retaining male clients in talk therapy. Understanding therapists' challenges when working with men can inform gender-specific training efforts. METHODS: Open-ended qualitative survey data were collected from a sample of 421 Australian-based therapists. Participants described that which they find most challenging about therapeutic work with men. Responses were analyzed using inductive thematic analysis. RESULTS: Three themes were revealed: (1) men's wavering commitment and engagement; (2) males as ill-equipped for therapy; and (3) therapists' uncertainty. Contrasting state and trait constructs, much of the men's state-based wavering commitment and engagement was positioned as amenable to change whereas traits assigned men as ill-equipped for therapy and unreachable. CONCLUSION: These findings underscore a clear need to better target training efforts to directly respond to the needs of therapists working with men, such that all therapists are well-equipped to meet men with gender-sensitive therapy.

Human complement receptor 1 (HuCR1) is a pivotal regulator of complement activity, acting on all three complement pathways as a membrane-bound receptor of C3b/C4b, C3/C5 convertase decay accelerator, and cofactor for factor I-mediated cleavage of C3b and C4b. In this study, we sought to identify a minimal soluble fragment of HuCR1, which retains the complement regulatory activity of the wildtype protein. To this end, we generated recombinant, soluble, and truncated versions of HuCR1 and compared their ability to inhibit complement activation in vitro using multiple assays. A soluble form of HuCR1, truncated at amino acid 1392 and designated CSL040, was found to be a more potent inhibitor than all other truncation variants tested. CSL040 retained its affinity to both C3b and C4b as well as its cleavage and decay acceleration activity and was found to be stable under a range of buffer conditions. Pharmacokinetic studies in mice demonstrated that the level of sialylation is a major determinant of CSL040 clearance in vivo. CSL040 also showed an improved pharmacokinetic profile compared with the full extracellular domain of HuCR1. The in vivo effects of CSL040 on acute complement-mediated kidney damage were tested in an attenuated passive antiglomerular basement membrane antibody-induced glomerulonephritis model. In this model, CSL040 at 20 and 60 mg/kg significantly attenuated kidney damage at 24 h, with significant reductions in cellular infiltrates and urine albumin, consistent with protection from kidney damage. CSL040 thus represents a potential therapeutic candidate for the treatment of complement-mediated disorders.

Regionally-based Australian men have a higher risk of suicide than those in urban centers, with similar trends observed internationally. Adopting a place-based approach to understanding men's suicide and harm prevention provides contextual insights to guide localised opportunities for the development of tailored gender-specific interventions. Men in rural Australia are typically portrayed as embodying idealized masculinity-dominant and tough, upholding strength and stoicism in the face of hardship. Such values can increase suicide risk in men by reducing help-seeking. The Macedon Ranges Shire is an inner regional municipality with a population of approximately 50,000 people spanning across 10 regional towns and surrounding farming areas in Victoria, Australia. Understanding the influence of masculinities on men's wellbeing and help seeking behaviours in a regional context is vital in order to inform effective local suicide prevention efforts. The present research involved in-depth qualitative interviews with 19 community stakeholders (M = 49.89 years, SD = 11.82) predominantly working in healthcare and community services including emergency services and education. Using thematic analysis, interview transcripts were coded and themes inductively derived. Stakeholders identified three key areas for understanding suicide risk and wellbeing for local men; 1) localizing masculinities, 2) belonging in community, and 3) engaging men. Findings illustrate that addressing men's wellbeing in regional areas requires a multifaceted whole-of-community approach. While diverse, local expressions of masculinities were seen as contributors to men's challenges understanding their emotional worlds and reticence for help-seeking. Of vital need is to provide diverse opportunities for men to connect with others in the region, and offer inclusive spaces where men feel accepted, welcomed and able to meaningfully contribute to the community. Not only will this assist by bolstering men's sense

OBJECTIVES: To develop consensus definitions for the core outcome set for pre-eclampsia. STUDY DESIGN: Potential definitions for individual core outcomes were identified across four formal definition development initiatives, nine national and international guidelines, 12 Cochrane systematic reviews, and 79 randomised trials. Eighty-six definitions were entered into the consensus development meeting. Ten healthcare professionals and three researchers, including six participants who had experience of conducting research in low- and middle-income countries, participated in the consensus development process. The final core outcome set was approved by an international steering group. RESULTS: Consensus definitions were developed for all core outcomes. When considering stroke, pulmonary oedema, acute kidney injury, raised liver enzymes, low platelets, birth weight, and neonatal seizures, consensus definitions were developed specifically for low- and middle-income countries because of the limited availability of diagnostic interventions including computerised tomography, chest x-ray, laboratory tests, equipment, and electroencephalogram monitoring. CONCLUSIONS: Consensus on measurements for the pre-eclampsia core outcome set will help to ensure consistency across future randomised trials and systematic reviews. Such standardization should make research evidence more accessible and facilitate the translation of research into clinical practice. Video abstract can be available at: www.dropbox.com/s/ftrgvrfu0u9glqd/6.%20Standardising%20definitions%20in%20teh%20pre-eclampsia%20core%20outcome%20set%3A%20a%20consensus%20development%20study.mp4?dl=0.

During haematopoiesis, haematopoietic stem cells differentiate into restricted potential progenitors before maturing into the many lineages required for oxygen transport, wound healing and immune response. We have updated Haemopedia, a database of gene-expression profiles from a broad spectrum of haematopoietic cells, to include RNA-seq gene-expression data from both mice and humans. The Haemopedia RNA-seq data set covers a wide range of lineages and progenitors, with 57 mouse blood cell types (flow sorted populations from healthy mice) and 12 human blood cell types. This data set has been made accessible for exploration and analysis, to researchers and clinicians with limited bioinformatics experience, on our online portal Haemosphere: https://www.haemosphere.org. Haemosphere also includes nine other publicly available high-quality data sets relevant to haematopoiesis. We have added the ability to compare gene expression across data sets and species by curating data sets with shared lineage designations or to view expression gene vs gene, with all plots available for download by the user.

PNPT1 (PNPase-polynucleotide phosphorylase) is involved in multiple RNA processing functions in the mitochondria. Bi-allelic pathogenic PNPT1 variants cause heterogeneous clinical phenotypes affecting multiple organs without any established genotype-phenotype correlations. Defects in PNPase can cause variable combined respiratory chain complex defects. Recently, it has been suggested that PNPase can lead to activation of an innate immune response. To better understand the clinical and molecular spectrum of patients with bi-allelic PNPT1 variants, we captured detailed clinical and molecular phenotypes of all 17 patients reported in the literature, plus seven new patients, including a 78-year-old male with the longest reported survival. A functional follow-up of genomic sequencing by cDNA studies confirmed a splicing defect in a novel, apparently synonymous, variant. Patient fibroblasts showed an accumulation of mitochondrial unprocessed PNPT1 transcripts, while blood showed an increased interferon response. Our findings suggest that functional analyses of the RNA processing function of PNPase are more sensitive than testing downstream defects in oxidative phosphorylation (OXPHPOS) enzyme activities. This research extends our knowledge of the clinical and functional consequences of bi-allelic pathogenic PNPT1 variants that may guide management and further efforts into understanding the pathophysiological mechanisms for therapeutic development.

A male neonate presented with severe weakness, hypotonia, contractures and congenital scoliosis. Skeletal muscle specimens showed marked atrophy and degeneration of fast fibers with striking nemaline rods and hypertrophy of slow fibers that were ultrastructurally normal. A neuromuscular gene panel identified a homozygous essential splice variant in TNNT3 (chr11:1956150G > A, NM_006757.3:c.681+1G > A). TNNT3 encodes skeletal troponin-Tfast and is associated with autosomal dominant distal arthrogryposis. TNNT3 has not previously been associated with nemaline myopathy (NM), a rare congenital myopathy linked to defects in proteins associated with thin filament structure and regulation. cDNA studies confirmed pathogenic consequences of the splice variant, eliciting exon-skipping and intron retention events leading to a frameshift. Western blot showed deficiency of troponin-Tfast protein with secondary loss of troponin-Ifast . We establish a homozygous splice variant in TNNT3 as the likely cause of severe congenital NM with distal arthrogryposis, characterized by specific involvement of Type-2 fibers and deficiency of troponin-Tfast .

MOTIVATION: Gene set enrichment (GSE) analysis allows researchers to efficiently extract biological insight from long lists of differentially expressed genes by interrogating them at a systems level. In recent years, there has been a proliferation of GSE analysis methods and hence it has become increasingly difficult for researchers to select an optimal GSE tool based on their particular dataset. Moreover, the majority of GSE analysis methods do not allow researchers to simultaneously compare gene set level results between multiple experimental conditions. RESULTS: The ensemble of genes set enrichment analyses (EGSEA) is a method developed for RNA-sequencing data that combines results from twelve algorithms and calculates collective gene set scores to improve the biological relevance of the highest ranked gene sets. EGSEA's gene set database contains around 25 000 gene sets from sixteen collections. It has multiple visualization capabilities that allow researchers to view gene sets at various levels of granularity. EGSEA has been tested on simulated data and on a number of human and mouse datasets and, based on biologists' feedback, consistently outperforms the individual tools that have been combined. Our evaluation demonstrates the superiority of the ensemble approach for GSE analysis, and its utility to effectively and efficiently extrapolate biological functions and potential involvement in disease processes from lists of differentially regulated genes. AVAILABILITY AND IMPLEMENTATION: EGSEA is available as an R package at http://www.bioconductor.org/packages/EGSEA/ . The gene sets collections are available in the R package EGSEAdata from http://www.bioconductor.org/packages/EGSEAdata/ . CONTACTS: monther.alhamdoosh@csl.com.au mritchie@wehi.edu.au. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.

Hematopoiesis is a multistage process involving the differentiation of stem and progenitor cells into distinct mature cell lineages. Here we present Haemopedia, an atlas of murine gene-expression data containing 54 hematopoietic cell types, covering all the mature lineages in hematopoiesis. We include rare cell populations such as eosinophils, mast cells, basophils, and megakaryocytes, and a broad collection of progenitor and stem cells. We show that lineage branching and maturation during hematopoiesis can be reconstructed using the expression patterns of small sets of genes. We also have identified genes with enriched expression in each of the mature blood cell lineages, many of which show conserved lineage-enriched expression in human hematopoiesis. We have created an online web portal called Haemosphere to make analyses of Haemopedia and other blood cell transcriptional datasets easier. This resource provides simple tools to interrogate gene-expression-based relationships between hematopoietic cell types and genes of interest.

© 2014, Springer-Verlag Berlin Heidelberg. Opportunistic oral infections caused by Candida albicans are frequent problems in immunocompromised patients. Management of such infections is limited due to the low number of antifungal drugs available, their relatively high toxicity and the emergence of antifungal resistance. Given these issues, our investigations have focused on novel derivatives of the antifungal antibiotic Nystatin A1, generated by modifications at the amino group of this molecule. The aims of this study were to evaluate the antifungal effectiveness and host cell toxicity of these new compounds using an in vitro model of oral candidosis based on a reconstituted human oral epithelium (RHOE). Initial studies employing broth microdilution, revealed that against planktonic C. albicans, Nystatin A1had lower minimal inhibitory concentration than novel derivatives. However, Nystatin A1was also markedly more toxic against human keratinocyte cells. Interestingly, using live/dead staining to assess C.albicans and tissue cell viability after RHOE infection, Nystatin A1derivatives were more active against Candida with lower toxicity to epithelial cells than the parent drug. Lactate dehydrogenase activity released by the RHOE indicated a fourfold reduction in tissue damage when certain Nystatin derivatives were used compared with Nystatin A1. Furthermore, compared with Nystatin A1, colonisation of the oral epithelium by C. albicans was notably reduced by the new polyenes. In the absence of antifungal agents, confocal laser scanning microscopy showed that C. albicans extensively invaded the RHOE. However, the presence of the novel derivatives greatly reduced or totally prevented this fungal invasion.

We describe a novel cloning method, referred to as insert-tagged (InTag) positive selection, for the rapid one-step reformatting of phage-displayed antibody fragments to full-length immunoglobulin Gs (IgGs). InTag positive selection enables recombinant clones of interest to be directly selected without cloning background, bypassing the laborious process of plating out cultures and colony screening and enabling the cloning procedure to be automated and performed in a high-throughput format. This removes a significant bottleneck in the functional screening of phage-derived antibody candidates and enables a large number of clones to be directly reformatted into IgG without the intermediate step of Escherichia coli expression and testing of soluble antibody fragments. The use of InTag positive selection with the Dyax Fab-on-phage antibody library is demonstrated, and optimized methods for the small-scale transient expression of IgGs at high levels are described. InTag positive selection cloning has the potential for wide application in high-throughput DNA cloning involving multiple inserts, markedly improving the speed and quality of selections from protein libraries.

Craniosynostosis is one of the most common craniofacial disorders encountered in clinical genetics practice, with an overall incidence of 1 in 2,500. Between 30% and 70% of syndromic craniosynostoses are caused by mutations in hotspots in the fibroblast growth factor receptor (FGFR) genes or in the TWIST1 gene with the difference in detection rates likely to be related to different study populations within craniofacial centers. Here we present results from molecular testing of an Australia and New Zealand cohort of 630 individuals with a diagnosis of craniosynostosis. Data were obtained by Sanger sequencing of FGFR1, FGFR2, and FGFR3 hotspot exons and the TWIST1 gene, as well as copy number detection of TWIST1. Of the 630 probands, there were 231 who had one of 80 distinct mutations (36%). Among the 80 mutations, 17 novel sequence variants were detected in three of the four genes screened. In addition to the proband cohort there were 96 individuals who underwent predictive or prenatal testing as part of family studies. Dysmorphic features consistent with the known FGFR1-3/TWIST1-associated syndromes were predictive for mutation detection. We also show a statistically significant association between splice site mutations in FGFR2 and a clinical diagnosis of Pfeiffer syndrome, more severe clinical phenotypes associated with FGFR2 exon 10 versus exon 8 mutations, and more frequent surgical procedures in the presence of a pathogenic mutation. Targeting gene hot spot areas for mutation analysis is a useful strategy to maximize the success of molecular diagnosis for individuals with craniosynostosis. © 2013 Wiley Periodicals, Inc.