Perceptions of bias in HIV prevention services, happiness with social support, and anticipated PrEP stigma among Black and Latine/x sexual and gender diverse individuals.

Although pre-exposure prophylaxis (PrEP) is a highly effective preventive treatment for HIV, anticipated PrEP stigma can hinder uptake. Perceptions of bias in HIV prevention and evaluations (e.g., happiness) tied to social support among Black and Latine/x sexual and gender diverse (SGD) individuals could be important correlates of anticipated PrEP stigma. To further this line of inquiry, a national sample of 872 Black and Latine/x SGD individuals who had and had never taken PrEP (Mage = 25.1, SD = 2.8) reported how they perceived HIV prevention and how happy they were with their social support. Multivariable linear regressions revealed that greater perceptions of bias in HIV prevention services were associated with higher anticipated PrEP stigma among Black and Latine/x SGD individuals who have never taken PrEP. Greater happiness with friend support was associated with lower PrEP stigma, whereas greater happiness with family support was associated with higher PrEP stigma among individuals who have taken PrEP. Findings highlight the need for PrEP and HIV interventions to address the intersectional stigma attached to prevention and for researchers to understand how evaluations of social support may contribute to stigma among Black and Latine/x SGD individuals.

Measurement invariance of the LGBT People of Color Microaggressions Scale among LGBTQ+ youth.

The LGBT People of Color Microaggressions Scale (LGBT-PCMS) is a widely used measure of intersectional microaggression experiences among sexual and gender minority people of color. Although it is widely used-and increasingly used in adolescent and young adult samples-it is unknown whether the LGBT-PCMS demonstrates similar measurement properties across subgroups of sexual and gender minority youth of color (SGMYOC). Among 4142 SGMYOC (ages 13-17) we found evidence for either partial or full scalar invariance (item loadings and intercepts were generally equal) across sexual orientation, race-ethnicity, and gender identity groups for all three subscales. Specific patterns of invariance and noninvariance across groups, as well as implications for the use of the LGBT-PCMS and its subscales among SGMYOC are discussed.

Stress of being outed to parents, LGBTQ family support, and depressive symptoms among sexual and gender diverse youth.

Limited scholarship has explored how a lack of agency in identity disclosure (being "outed") to parents is associated with mental health experiences of sexual and gender diverse youth (SGDY). With a national sample of SGDY (N = 9272; 66.8% White non-Hispanic) aged 13-17 (Mage = 15.63, SD = 1.24), this study first compared social position differences between SGDY who were outed to their parents compared to those not outed, and second, investigated how the stress from being outed to parents was associated with LGBTQ family support and depressive symptoms. Results revealed that SGDY who were outed to their parents reported higher levels of depressive symptoms and lower amounts of LGBTQ family support than SGDY who were not outed to their parents. In addition, greater stress from being outed to parents was indirectly associated with higher depressive symptoms through lower LGBTQ family support. These relationships significantly varied across gender identity. Findings highlight the importance of instilling greater agency in disclosure experiences among SGDY.

Sexual Identity Disclosure and Alcohol Experiences Among LGBTQ+ Adolescents.

Sexual and gender minority youth (SGMY) report greater alcohol use in comparison to their heterosexual counterparts. Prior research has found that elevated alcohol use among SGMY can be explained by minority stress experiences. Sexual identity outness may be another factor that drives alcohol use among SGMY, given that outness is associated with alcohol use among older sexual and gender minority samples. We examined how patterns of sexual identity outness were associated with lifetime alcohol use, past-30-day alcohol use, and past-30-day heavy episodic drinking. Data were drawn from the LGBTQ National Teen Survey (N = 8884). Participants were SGMY aged 13 to 17 (mean age = 15.59) years living in the US. Latent class analysis was used to identify sexual identity outness patterns. Multinomial regressions were used to examine the probability of class membership by alcohol use. Six outness classes were identified: out to all but teachers (n = 1033), out to siblings and peers (n = 1808), out to siblings and LGBTQ+ peers (n = 1707), out to LGBTQ+ peers (n = 1376), mostly not out (n = 1653), and very much not out (n = 1307). SGMY in classes characterized by greater outness to peers, friends, and family had greater odds of lifetime alcohol use compared with SGMY in classes characterized by lower outness. These findings suggest that SGMY with greater sexual identity outness may be a target for alcohol use prevention programming. Differences in sexual identity outness may be explained by minority stress factors.

Sexual minority youth in romantic relationships: Associations with youth well-being.

This study investigated differences in depressive symptoms, loneliness, and self-esteem for monosexual (lesbian, gay) and plurisexual (bisexual, pansexual, queer) sexual minority youth (SMY) by relationship status (single, partnered) and relationship configuration (same-gender partner, different-gender partner). Participants included 338 SMY (Mage = 19.10 years) who reported on their relationship status, partner's gender identity, well-being, and ability to confide in partner about LGBTQ issues. Results indicated that for plurisexual youth, single status was associated with greater loneliness; plurisexual youth with same-gender partners reported fewer depressive symptoms and marginally greater ability to confide in their partner about LGBTQ issues than those with different-gender partners. Findings reveal similarities across SMY while also highlighting some unique challenges among plurisexual youth with different-gender partners.

Youth-specific sexual and gender minority state-level policies: Implications for pronoun, name, and bathroom/locker room use among gender minority youth.

While research that investigates the importance of school-level promotive factors (e.g., teacher support) for sexual and gender minority youth (SGMY) well-being has proliferated, less research has focused on state-level climate and policy implications for gender minority youth-specific experiences. This study investigated the impact of two youth-specific SGM state-level laws (i.e., "anti-LGBT laws" and conversion therapy bans) on social transition experiences (i.e., name/pronoun use and using desired bathroom/locker rooms) of GMY (n = 4000) aged 13-17. Through a series of multivariable regression models, it was determined that the absence of laws that restricted rights for sexual and gender minority people was associated with greater use of the correct name and correct pronouns for transgender youth. These differences were further explained by binary gender identity (transgender binary or nonbinary) status, region, and age in multivariable models. Findings highlight the importance of enacting more uniform protections for SGMY, especially to protect transgender youth that live in the southern region of the U.S.

Contextualizing the Well-being of Asexual Youth: Evidence of Differences in Family, Health, and School Outcomes.

Despite increasing efforts to better understand sexual and gender minority youth (SGMY), asexual youth remain understudied. This study examines differences in health, family support, and school safety among asexual youth (n = 938) from a national study of SGMY (N = 17,112) ages 13-17. Compared to non-asexual youth, asexual youth were more likely to identify as transgender and report a disability, and less likely to identify as Black or Hispanic/Latino. Transgender (versus cisgender) asexual youth fared worse on most study outcomes. Cisgender asexual (versus cisgender non-asexual) youth fared worse on all study outcomes. Transgender asexual (versus transgender non-asexual) youth reported lower sexuality-related family support. These findings underscore the role of gender identity in understanding the experiences of asexual youth.

mRNA and microRNA analysis reveals modulation of biochemical pathways related to addiction in the ventral tegmental area of methamphetamine self-administering rats.

BACKGROUND: Methamphetamine is a highly addictive central nervous system stimulant with increasing levels of abuse worldwide. Alterations to mRNA and miRNA expression within the mesolimbic system can affect addiction-like behaviors and thus play a role in the development of drug addiction. While many studies have investigated the effects of high-dose methamphetamine, and identified neurotoxic effects, few have looked at the role that persistent changes in gene regulation play following methamphetamine self-administration. Therefore, the aim of this study was to identify RNA changes in the ventral tegmental area following methamphetamine self-administration. We performed microarray analyses on RNA extracted from the ventral tegmental area of Sprague-Dawley rats following methamphetamine self-administration training (2 h/day) and 14 days of abstinence. RESULTS: We identified 78 miRNA and 150 mRNA transcripts that were differentially expressed (fdr adjusted p < 0.05, absolute log2 fold change >0.5); these included genes not previously associated with addiction (miR-125a-5p, miR-145 and Foxa1), loci encoding receptors related to drug addiction behaviors and genes with previously recognized roles in addiction such as miR-124, miR-181a, DAT and Ret. CONCLUSION: This study provides insight into the effects of methamphetamine on RNA expression in a key brain region associated with addiction, highlighting the possibility that persistent changes in the expression of genes with both known and previously unknown roles in addiction occur.

Serum bilirubin concentration is modified by UGT1A1 haplotypes and influences risk of type-2 diabetes in the Norfolk Island genetic isolate.

BACKGROUND: Located in the Pacific Ocean between Australia and New Zealand, the unique population isolate of Norfolk Island has been shown to exhibit increased prevalence of metabolic disorders (type-2 diabetes, cardiovascular disease) compared to mainland Australia. We investigated this well-established genetic isolate, utilising its unique genomic structure to increase the ability to detect related genetic markers. A pedigree-based genome-wide association study of 16 routinely collected blood-based clinical traits in 382 Norfolk Island individuals was performed. RESULTS: A striking association peak was located at chromosome 2q37.1 for both total bilirubin and direct bilirubin, with 29 SNPs reaching statistical significance (P < 1.84 × 10(-7)). Strong linkage disequilibrium was observed across a 200 kb region spanning the UDP-glucuronosyltransferase family, including UGT1A1, an enzyme known to metabolise bilirubin. Given the epidemiological literature suggesting negative association between CVD-risk and serum bilirubin we further explored potential associations using stepwise multivariate regression, revealing significant association between direct bilirubin concentration and type-2 diabetes risk. In the Norfolk Island cohort increased direct bilirubin was associated with a 28% reduction in type-2 diabetes risk (OR: 0.72, 95% CI: 0.57-0.91, P = 0.005). When adjusted for genotypic effects the overall model was validated, with the adjusted model predicting a 30% reduction in type-2 diabetes risk with increasing direct bilirubin concentrations (OR: 0.70, 95% CI: 0.53-0.89, P = 0.0001). CONCLUSIONS: In summary, a pedigree-based GWAS of blood-based clinical traits in the Norfolk Island population has identified variants within the UDPGT family directly associated with serum bilirubin levels, which is in turn implicated with reduced risk of developing type-2 diabetes within this population.

Methylation differences at the HLA-DRB1 locus in CD4+ T-Cells are associated with multiple sclerosis.

BACKGROUND: Multiple sclerosis (MS) is thought to be caused by T-cell mediated autoimmune dysfunction. Risk of developing MS is influenced by environmental and genetic factors. Modifiable differences in DNA methylation are recognized as epigenetic contributors to MS risk and may provide a valuable link between environmental exposure and inherited genetic systems. OBJECTIVES AND METHODS: To identify methylation changes associated with MS, we performed a genome-wide DNA methylation analysis of CD4+ T cells from 30 patients with relapsing-remitting MS and 28 healthy controls using Illumina 450K methylation arrays. RESULTS: A striking differential methylation signal was observed at chr. 6p21, with a peak signal at HLA-DRB1. After prioritisation, we identified a panel of 74 CpGs associated with MS in this cohort. Most notably we found evidence of a major effect CpG island in DRB1 in MS cases (pFDR < 3 × 10(-3)). In addition, we found 55 non-HLA CpGs that exhibited differential methylation, many of which localise to genes previously linked to MS. CONCLUSIONS: Our findings provide the first evidence for association of DNA methylation at HLA-DRB1 in relation to MS risk. Further studies are now warranted to validate and understand how these findings are involved in MS pathology.

Experiences of stress and social safety among sexual and gender minority youth by disability status.

BACKGROUND: Though separate bodies of research have shown sexual and gender minority (SGM) youth, and youth with disabilities, separately, face distinct social and health disparities, little is known about youth who both identify as SGM and have disabilities. OBJECTIVE: The current study examined differences in wellbeing among SGM youth by disability category (i.e., physical, developmental, psychiatric) across victimization, bullying, dating violence, school safety, and experienced stress. METHODS: Using self-reported data from 9418 SGM youth aged 13-17 in the United States, multivariate linear regressions were conducted to examine how stress and social safety experiences varied across disability status. RESULTS: Compared to SGM youth without a disability, SGM youth across all disability categories (physical, developmental, psychiatric) had greater odds of LGBT- and disability-based victimization, greater average stress, as well as lower levels of school safety. SGM youth with any disability, physical disability, or psychiatric disability also had greater odds of dating violence compared to SGM youth without a disability. CONCLUSION: SGM youth with disabilities may be in particular need of targeted programs that address both disability and sexual/gender identities, and may benefit from increased supports across developmental contexts (e.g., against bullying in school). Stakeholders should consider how such support can be improved, tailored, and implemented, for SGM youth and the diversity of disabilities they have.

Examining Mental Health and Bullying Concerns at the Intersection of Sexuality, Gender, Race, and Ethnicity Among a National Sample of Sexual and Gender Diverse Youth.

Purpose: Most extant scholarship that examines the health experiences of sexual and gender diverse youth (SGDY) is limited in the ability to apply an intersectional framework due to small sample sizes and limitations in analytic methods that only analyze the independent contribution of social identities. To address this gap, this study explored the well-being of youth at the intersection of ethnic, racial, sexual, and gender identities in relation to mental health and bullying. Methods: Data were from a U.S. national survey of SGDY aged 13-18 years, collected in 2022 (N = 12,822). Exhaustive Chi-square Automatic Interaction Detection analysis identified intersectional social positions bearing the greatest burden of negative health-related experiences (depression, anxiety, and past 30-day in-person victimization). Results: Transgender boys were among those at the highest prevalence for compromised mental health and peer-based in-person victimization. Although the primary distinguishing factor was transgender identity for depression and anxiety, there were no racial/ethnic distinctions, corroborating some previous scholarship. Asian cisgender and transgender girl SGDY shared the lowest burden of peer-based in-person victimization in school. Conclusion: Our findings suggest a need for scholars, health professionals, and other stakeholders to better understand the mechanisms that drive negative health experiences and in-person victimization experiences at the intersections of sexual, gender, racial, and ethnic identities.

Cognitive and neuroimaging outcomes in individuals with benign and low-grade brain tumours receiving radiotherapy: a protocol for a prospective cohort study.

INTRODUCTION: Radiation-induced cognitive decline (RICD) occurs in 50%-90% of adult patients 6 months post-treatment. In patients with low-grade and benign tumours with long expected survival, this is of paramount importance. Despite advances in radiation therapy (RT) treatment delivery, better understanding of structures important for RICD is necessary to improve cognitive outcomes. We hypothesise that RT may affect network topology and microstructural integrity on MRI prior to any gross anatomical or apparent cognitive changes. In this longitudinal cohort study, we aim to determine the effects of RT on brain structural and functional integrity and cognition. METHODS AND ANALYSIS: This study will enroll patients with benign and low-grade brain tumours receiving partial brain radiotherapy. Patients will receive either hypofractionated (>2 Gy/fraction) or conventionally fractionated (1.8-2 Gy/fraction) RT. All participants will be followed for 12 months, with MRIs conducted pre-RT and 6-month and 12 month post-RT, along with a battery of neurocognitive tests and questionnaires. The study was initiated in late 2018 and will continue enrolling through 2024 with final follow-ups completing in 2025. The neurocognitive battery assesses visual and verbal memory, attention, executive function, processing speed and emotional cognition. MRI protocols incorporate diffusion tensor imaging and resting state fMRI to assess structural connectivity and functional connectivity, respectively. We will estimate the association between radiation dose, imaging metrics and cognitive outcomes. ETHICS AND DISSEMINATION: This study has been approved by the Research Subjects Review Board at the University of Rochester (STUDY00001512: Cognitive changes in patients receiving partial brain radiation). All results will be published in peer-reviewed journals and at scientific conferences. TRIAL REGISTRATION NUMBER: ClinicalTrials.gov NCT04390906.

MS0621, a novel small-molecule modulator of Ewing sarcoma chromatin accessibility, interacts with an RNA-associated macromolecular complex and influences RNA splicing.

Ewing sarcoma is a cancer of children and young adults characterized by the critical translocation-associated fusion oncoprotein EWSR1::FLI1. EWSR1::FLI1 targets characteristic genetic loci where it mediates aberrant chromatin and the establishment of de novo enhancers. Ewing sarcoma thus provides a model to interrogate mechanisms underlying chromatin dysregulation in tumorigenesis. Previously, we developed a high-throughput chromatin-based screening platform based on the de novo enhancers and demonstrated its utility in identifying small molecules capable of altering chromatin accessibility. Here, we report the identification of MS0621, a molecule with previously uncharacterized mechanism of action, as a small molecule modulator of chromatin state at sites of aberrant chromatin accessibility at EWSR1::FLI1-bound loci. MS0621 suppresses cellular proliferation of Ewing sarcoma cell lines by cell cycle arrest. Proteomic studies demonstrate that MS0621 associates with EWSR1::FLI1, RNA binding and splicing proteins, as well as chromatin regulatory proteins. Surprisingly, interactions with chromatin and many RNA-binding proteins, including EWSR1::FLI1 and its known interactors, were RNA-independent. Our findings suggest that MS0621 affects EWSR1::FLI1-mediated chromatin activity by interacting with and altering the activity of RNA splicing machinery and chromatin modulating factors. Genetic modulation of these proteins similarly inhibits proliferation and alters chromatin in Ewing sarcoma cells. The use of an oncogene-associated chromatin signature as a target allows for a direct approach to screen for unrecognized modulators of epigenetic machinery and provides a framework for using chromatin-based assays for future therapeutic discovery efforts.

Construction of a cytosolic firefly luciferase reporter cassette for use in PCR-mediated gene deletion and fusion in Saccharomyces cerevisiae.

Monitoring promoter response to environmental changes using reporter systems has provided invaluable information regarding cellular state. With the development of in vivo luciferase reporter systems, inexpensive, sensitive and accurate promoter assays have been developed without the variability reported between in vitro samplings. Current luciferase reporter systems, however, are largely inflexible to modifications to the promoter of interest. To overcome problems in flexibility and stability of these expression vectors, we report the creation of a novel vector system which introduces a cytosol-localized Photinus pyralis luciferase [LUC*(-SKL)] capable of one-step, in vivo measurements into a promoter-reporter system via PCR-based gene deletion and fusion. After introduction of the reporter under HUG1 promoter control, cytosolic localization was confirmed by fluorescence microscopy. The dose-response of this novel construct was then compared with that of a similar HUG1Δ::yEGFP1 promoter-reporter system and shown to give a similar response pattern.

Homogeneous detection of cyanobacterial DNA via polymerase chain reaction.

AIMS: To design a primer set enabling the identification through PCR of high-quality DNA for routine and high-throughput genomic screening of a diverse range of cyanobacteria. METHODS AND RESULTS: A codon-equivalent multiple alignment of the phycocyanin alpha-subunit coding sequence (cpcA) of 22 cyanobacteria was generated and analysed to produce a single degeneracy primer set with virtually uniform product size. Also, an 18S ribosomal RNA detection set is proposed for rejecting false positives. The primer sets were tested against five diverse cyanobacteria, Chlorella vulgaris, Saccharomyces cerevisiae, and Escherichia coli. All five cyanobacteria showed positive amplification of cpcA product with homogeneous fragment length, and no products were observed for any other organism. Additionally, the only product formation observed for the 18S rRNA set was in C. vulgaris and S. cerevisiae. CONCLUSIONS: The newly proposed primer set served as effective check primers for cyanobacteria. Cyanobacteria gDNA had a positive, homogenous result, while other bacteria, eukaryotes and alga tested were negative. SIGNIFICANCE AND IMPACT OF THE STUDY: These novel, broad-spectrum primers will greatly increase the utility of PCR on newly discovered cyanobacterial species.

Pedigree derived mutation rate across the entire mitochondrial genome of the Norfolk Island population.

Estimates of mutation rates for various regions of the human mitochondrial genome (mtGenome) vary widely, depending on whether they are inferred using a phylogenetic approach or obtained directly from pedigrees. Traditionally, only the control region, or small portions of the coding region have been targeted for analysis due to the cost and effort required to produce whole mtGenome Sanger profiles. Here, we report one of the first pedigree derived mutation rates for the entire human mtGenome. The entire mtGenome from 225 individuals originating from Norfolk Island was analysed to estimate the pedigree derived mutation rate and compared against published mutation rates. These individuals were from 45 maternal lineages spanning 345 generational events. Mutation rates for various portions of the mtGenome were calculated. Nine mutations (including two transitions and seven cases of heteroplasmy) were observed, resulting in a rate of 0.058 mutations/site/million years (95% CI 0.031-0.108). These mutation rates are approximately 16 times higher than estimates derived from phylogenetic analysis with heteroplasmy detected in 13 samples (n = 225, 5.8% individuals). Providing one of the first pedigree derived estimates for the entire mtGenome, this study provides a better understanding of human mtGenome evolution and has relevance to many research fields, including medicine, anthropology and forensics.

Is evolutionary history repeatedly rewritten in light of new fossil discoveries?

Mass media and popular science journals commonly report that new fossil discoveries have 'rewritten evolutionary history'. Is this merely journalistic hyperbole or is our sampling of systematic diversity so limited that attempts to derive evolutionary history from these datasets are premature? We use two exemplars-catarrhine primates (Old World monkeys and apes) and non-avian dinosaurs-to investigate how the maturity of datasets can be assessed. Both groups have been intensively studied over the past 200 years and so should represent pinnacles in our knowledge of vertebrate systematic diversity. We test the maturity of these datasets by assessing the completeness of their fossil records, their susceptibility to changes in macroevolutionary hypotheses and the balance of their phylogenies through study time. Catarrhines have shown prolonged stability, with discoveries of new species being evenly distributed across the phylogeny, and thus have had little impact on our understanding of their fossil record, diversification and evolution. The reverse is true for dinosaurs, where the addition of new species has been non-random and, consequentially, their fossil record, tree shape and our understanding of their diversification is rapidly changing. The conclusions derived from these analyses are relevant more generally: the maturity of systematic datasets can and should be assessed before they are exploited to derive grand macroevolutionary hypotheses.

Muscle Loss is Associated with Risk of Orthostatic Hypotension in Older Men and Women.

BACKGROUND: Muscle provides a reservoir for water to maintain fluid volume and blood pressure, so older adults may be at risk for orthostatic hypotension due to muscle loss with age. OBJECTIVES: To evaluate the association between muscle loss with age and postural blood pressure. DESIGN: Longitudinal comparison of overnight changes in hydration, postural blood pressure, and strength. SETTING: Community field study. PARTICIPANTS: Sixty-nine men and women (76.0 ± 0.8 years) with low (Low) or normal (Normal) muscle based on the Lean Mass Index. MEASUREMENTS: Body composition was measured with bioelectrical impedance analysis. Postural blood pressure was measured sequentially (lying, sitting, standing). Strength was measured with a handgrip dynamometer, Arm Curl test, and Chair Stand test. RESULTS: On Day 1, Low had less hydration and a significant drop in postural systolic blood pressure compared to Normal (lying to standing: -11.06 ± 2.36 vs. +1.14 ± 2.20 mmHg, p < 0.001). Overnight, both groups lost significant total body water, while fluid volume was unchanged. On Day 2, both groups experienced significant drops in postural systolic blood pressure, although the drop in Low was more profound and significantly greater than Normal (lying to standing: -16.85 ± 2.50 vs. -3.89 ± 2.52 mmHg, p = 0.001). On both days, Normal compensated for postural changes with increases in postural diastolic blood pressure not observed in Low. Only Low experienced significant overnight decreases in all strength measures. CONCLUSIONS: In older men and women, muscle loss with age is accompanied by loss of hydration and less stable early morning postural systolic blood pressure that increase risk for orthostatic hypotension and can also increase risk for falls.

Phylogenetically structured variance in felid bite force: the role of phylogeny in the evolution of biting performance.

A key question in evolution is the degree to which morphofunctional complexes are constrained by phylogeny. We investigated the role of phylogeny in the evolution of biting performance, quantified as bite forces, using phylogenetic eigenvector regression. Results indicate that there are strong phylogenetic signals in both absolute and size-adjusted bite forces, although it is weaker in the latter. This indicates that elimination of size influences reduces the level of phylogenetic inertia and that the majority of the phylogenetic constraint is a result of size. Tracing the evolution of bite force through phylogeny by character optimization also supports this notion, in that relative bite force is randomly distributed across phylogeny whereas absolute bite force diverges according to clade. The nonphylogenetically structured variance in bite force could not be sufficiently explained by species-unique morphology or by ecology. This study demonstrates the difficulties in identifying causes of nonphylogenetically structured variance in morphofunctional character complexes.