Unraveling the biology of a fungal meningitis pathogen using chemical genetics.

The fungal meningitis pathogen Cryptococcus neoformans is a central driver of mortality in HIV/AIDS. We report a genome-scale chemical genetic data map for this pathogen that quantifies the impact of 439 small-molecule challenges on 1,448 gene knockouts. We identified chemical phenotypes for 83% of mutants screened and at least one genetic response for each compound. C. neoformans chemical-genetic responses are largely distinct from orthologous published profiles of Saccharomyces cerevisiae, demonstrating the importance of pathogen-centered studies. We used the chemical-genetic matrix to predict novel pathogenicity genes, infer compound mode of action, and to develop an algorithm, O2M, that predicts antifungal synergies. These predictions were experimentally validated, thereby identifying virulence genes, a molecule that triggers G2/M arrest and inhibits the Cdc25 phosphatase, and many compounds that synergize with the antifungal drug fluconazole. Our work establishes a chemical-genetic foundation for approaching an infection responsible for greater than one-third of AIDS-related deaths.

Retinoic acid, an essential component of the roof plate organizer, promotes the spatiotemporal segregation of dorsal neural fates.

Dorsal neural tube-derived retinoic acid promotes the end of neural crest production and transition into a definitive roof plate. Here, we analyze how this impacts the segregation of central and peripheral lineages, a process essential for tissue patterning and function. Localized in ovo inhibition in quail embryos of retinoic acid activity followed by single-cell transcriptomics unraveled a comprehensive list of differentially expressed genes relevant to these processes. Importantly, progenitors co-expressed neural crest, roof plate and dI1 interneuron markers, indicating a failure in proper lineage segregation. Furthermore, separation between roof plate and dI1 interneurons is mediated by Notch activity downstream of retinoic acid, highlighting their crucial role in establishing the roof plate-dI1 boundary. Within the peripheral branch, where absence of retinoic acid resulted in neural crest production and emigration extending into the roof plate stage, sensory progenitors failed to separate from melanocytes, leading to formation of a common glia-melanocyte cell with aberrant migratory patterns. In summary, the implementation of single-cell RNA sequencing facilitated the discovery and characterization of a molecular mechanism responsible for the segregation of dorsal neural fates during development.

Transformational narrative changes as a community-level approach to the prevention of adverse childhood experiences and substance use.

There is increasing scientific evidence linking substance use, childhood adversity, and social determinants of health. However, little research has considered the evaluation of community-level strategies to reduce substance use by increasing awareness and implementation of evidence-based strategies for preventing adverse childhood experiences (ACEs). This article lays out the conceptual framework for a $2.9 million demonstration project designed to raise awareness of the impact of ACEs on substance use, including primary prevention strategies. Communities used transformational narrative change-with an emphasis on the voices of those most impacted by ACEs and substance use-to highlight the importance of addressing social determinants of health along with primary prevention strategies. The conceptual background highlighted in this article informed media, public health, and local efforts in the three accompanying articles and invited commentary. These findings may help inform future efforts to promote community-level strategies and strengthen the evidence-base for transformational narrative change efforts.

Increasing support for the prevention of adverse childhood experiences and substance use: Implementation of narrative change strategies in local health departments.

Adverse childhood experiences (ACEs) are potentially traumatic but preventable experiences that occur before the ages of 18, including child abuse, witnessing violence, and parental substance use. ACEs have been linked with increased risk for substance use, along with a variety of other negative health outcomes. However, there is limited evidence of community-level strategies that link ACEs and substance to increase awareness of prevention efforts. This article reports on a $2.9 million program to promote health equity and inform narratives for the prevention of ACEs and substance use within three Midwestern communities. Program partners sought to create new transformational narratives that linked ACEs and substance use, while underscoring the importance of addressing social determinants of health (SDOH) that lead to disparities in ACEs and substance use. A mixed-methods evaluation design included document review, in-depth interviews with program staff (N = 8) and community liaisons (N = 2), and site reports from program staff (N = 8) and their community partners (N = 17). Analyses showed that successful implementation efforts had early leadership buy-in and support, set clear and manageable expectations at the outset of implementation, and developed strong relationships with organizations that engage in health equity work. Training and technical assistance were critical to helping community partners build trust, recognize each other's perspectives, broaden and reframe their world view, and better understand narrative efforts for the primary prevention of ACEs and substance use.

Does binge drinking mediate the relationship between four adverse childhood experiences and adult traumatic brain injury? Results from the National Longitudinal Survey of Youth 1979 Cohort.

OBJECTIVE: Adverse childhood experiences (ACEs) are associated with increased risk of sustaining a traumatic brain injury (TBI). Alcohol use may play an important role in this relationship. This study examines whether binge drinking mediates the relationship between four ACEs and TBIs sustained in adulthood. METHODS: Using the National Longitudinal Survey of Youth, 1979 cohort, we conducted longitudinal mediation analyses (n=6317). Interviews occurred annually from 1979 to 1994 and biennially until 2016. We evaluated the direct and indirect effects of individual ACEs (ie, experiencing physical violence, low parental warmth, familial alcoholism and familial mental illness; reported retrospectively) and a cumulative ACEs score on mean level of binge drinking (calculated across waves) and having a TBI in adulthood. To establish temporality, we included binge drinking that was measured at age 18 or older and before any reported TBI. RESULTS: Cumulative ACEs, familial alcoholism and physical abuse exposure were significantly associated with having a TBI through binge drinking, although this only explained a small part of the association between ACEs and TBI. Other ACEs were not significantly associated with binge drinking or TBI. CONCLUSION: The results indicate that while ACEs and adult TBI risk were significantly associated, lifetime binge drinking explains only a small part of the association. Future research could examine alternative social, biological and behavioural mechanisms along the pathway between ACEs and TBI. Determining this mechanism will allow public health practitioners to design and implement effective TBI prevention programmes for those at higher risk of injury due to ACE exposure.

The role of gender and romantic jealousy in intimate partner violence against women, a mixed-methods study in Northern Ecuador.

Male romantic jealousy is a commonly cited driver of intimate partner violence against women. An in-depth, contextualised understanding of the pathways and mechanisms from jealousy to intimate partner violence is, however, needed to inform programmes and interventions. We triangulated data from 48 interviews, eight focus groups and 1216 survey findings from low-income married women and men in northern Ecuador. Male jealousy was associated with controlling behaviours (aOR: 14.47, 95% CI: 9.47, 22.12) and sexual intimate partner violence (aOR: 2.4, 95% CI: 1.12, 5.12). Controlling behaviours were associated with physical and sexual intimate partner violence (aOR: 2.16, 95% CI: 1.21, 3.84). Qualitatively we found that most respondents framed jealousy within a discourse of love, and three triggers of male jealousy leading to intimate partner violence were identified: (1) community gossip, which acted as a mechanism of community control over women's movements and sexuality; (2) women joining the labour force, which was quantitatively associated with intimate partner violence and partially mediated by jealousy; and (3) women's refusal to have sex, which could lead husbands to coerce sex through accusations of infidelity. Gender-transformative interventions at the individual, couple and community level providing models of alternative masculinities and femininities may offer promise in reducing intimate partner violence in Ecuador. Importantly, future economic empowerment interventions should address jealousy to mitigate potential intimate partner violence backlash.

Examining narratives around adverse childhood experiences and social determinants of health in media coverage of substance use in two mid-western cities.

Local media narratives play an important role in how people interpret and propose solutions for health issues in their community. This research characterized narratives about adverse childhood experiences (ACEs), and/or social determinants of health (SDOH) in media coverage of substance use. Scans covered articles published in the Detroit Free Press and the Cincinnati Enquirer from March 1, 2019 to June 1, 2019 and March 1, 2021 to June 1, 2021. Scans used search terms for opioids and substance use. Included articles were coded and analyzed for narratives about why people use substances, how to prevent substance use, and how ACEs or SDOH relate to substance use. While half of the included articles reported on the overdose epidemic, the most common type of media coverage reported on criminal justice milestones. Other common narratives identified addiction as an illness that should be treated; and over-prescription of painkillers or the strength of the drugs as causes of substance use disorders. Narratives about SDOH and the primary prevention of ACEs and substance use were limited. Transformational narrative change work can increase support for addressing the root causes of ACEs and substance use. Results suggest this strategy remains largely untapped in the formal media.

Notch signaling is a critical initiator of roof plate formation as revealed by the use of RNA profiling of the dorsal neural tube.

BACKGROUND: The dorsal domain of the neural tube is an excellent model to investigate the generation of complexity during embryonic development. It is a highly dynamic and multifaceted region being first transiently populated by prospective neural crest (NC) cells that sequentially emigrate to generate most of the peripheral nervous system. Subsequently, it becomes the definitive roof plate (RP) of the central nervous system. The RP, in turn, constitutes a patterning center for dorsal interneuron development. The factors underlying establishment of the definitive RP and its segregation from NC and dorsal interneurons are currently unknown. RESULTS: We performed a transcriptome analysis at trunk levels of quail embryos comparing the dorsal neural tube at premigratory NC and RP stages. This unraveled molecular heterogeneity between NC and RP stages, and within the RP itself. By implementing these genes, we asked whether Notch signaling is involved in RP development. First, we observed that Notch is active at the RP-interneuron interface. Furthermore, gain and loss of Notch function in quail and mouse embryos, respectively, revealed no effect on early NC behavior. Constitutive Notch activation caused a local downregulation of RP markers with a concomitant development of dI1 interneurons, as well as an ectopic upregulation of RP markers in the interneuron domain. Reciprocally, in mice lacking Notch activity, both the RP and dI1 interneurons failed to form and this was associated with expansion of the dI2 population. CONCLUSIONS: Collectively, our results offer a new resource for defining specific cell types, and provide evidence that Notch is required to establish the definitive RP, and to determine the choice between RP and interneuron fates, but not the segregation of RP from NC.

Breaking the cycle of Adverse Childhood Experiences (ACEs): Economic position moderates the relationship between mother and child ACE scores among Black and Hispanic families.

IMPORTANCE: Adverse Childhood Experiences (ACEs) are prevalent, preventable, and a public health issue that cycles from one generation to the next with serious implications for health and wellbeing, particularly. Research is needed to identify factors, including those related to economic position (i.e., wage, net family wealth, home ownership), that break the cycle of ACEs and inform decisions about policies, practices, and programs. OBJECTIVE: To determine whether economic position moderates the association between mother's ACE score and child's ACE score and whether these pathways differ by race and ethnicity. DESIGN: Conducted regression and moderation analysis using mother-child dyadic data from panel surveys, stratified by race. The simple slopes for the interactions were probed to determine the magnitude and significance of the interaction. SETTING: Secondary data analysis utilizing data from two cohorts of the National Longitudinal Surveys: 1) National Longitudinal Survey of Youth 1979; and 2) National Longitudinal Survey of Youth 1979 Children and Young Adults. PARTICIPANTS: The sample included 6,261 children and 2,967 matched mothers. MAIN OUTCOMES S AND MEASURES: The outcome variable was the child's ACE score. Mother's ACE score was the independent variable. Three economic position moderators were examined: mother's and her spouse's average wage and salary, average net family wealth, and percent of time owning a home during her child's first five years of life. RESULTS: Mother's ACE score was positively associated with her child's ACE score. Economic position was a significant moderator for Black families. Higher wages and net family wealth during children's first five years were associated with weakened associations between mother and child ACEs for Black families. For Hispanic families, higher wages and salary were significantly associated with weakened associations. Among White families, higher net family wealth was associated with stronger ACEs transmission. CONCLUSIONS AND RELEVANCE: Taken together, these findings highlight the important role that economic position may play on breaking the cycle of ACEs. This information can inform decisions about what public assistance policies, practices, and programs may be used to improve economic stability among families as an effective ACEs prevention strategy, and for whom these strategies might be most effective at reducing the cycle of ACEs.

Fostering gender equality and alternatives to violence: perspectives on a gender-transformative community mobilisation programme in rural South Africa.

Gender-based violence and violence against children are significant problems in South Africa. Community mobilisation and gender-transformative programming are promising approaches to address and reduce violence. A quantitative evaluation of One Man Can, a gender-transformative community mobilisation programme in South Africa, found mixed results in increasing gender-equitable behaviours and reducing violence. To better understand these findings, we analyse longitudinal qualitative data from community mobilisers, community members and community action teams, exploring individual and community-level factors that facilitate and hinder change. Interviews and focus groups were transcribed and analysed. Participants self-reported changes in their gender-equitable attitudes and use of violence as a result of participation in the programme, although some participants also reported opposition to shifting to a more gender-equitable culture. Facilitators to change included the internalisation of gender-transformative messaging and supportive social networks, which was buoyed by a shared vocabulary in their community generated by One Man Can. Because the programme targeted a critical mass of community members with gender-transformative programming, mobilisers and community action teams were held accountable by community members to model non-violent behaviour. Results reinforce the importance of addressing facilitators and barriers to change at both individual and community levels.