Youth Participatory Action Communication Research: A Model for Developing Youth-Driven Health Campaigns.

Youth offer valuable insight on health communication needs and solutions in their communities. We propose youth participatory action communication research (YPACR) as a model for health campaign development that engages youth perspectives in applying systematic theory-informed communication research to addressing youth-identified health priorities. YPACR informed a series of paid high school internship programs in West Philadelphia, in which youth interns identified mental health help-seeking communication as a need among peers. In Phase 1, guided by the reasoned action approach and Hornik & Woolf method, youth interns conducted a survey measuring behavioral beliefs, normative beliefs, and control beliefs associated with mental health help-seeking, as well as trusted sources of mental health information, among local high school students. Survey results suggested control (self-efficacy) was an important message target and peers were trusted mental health information sources. In Phase 2, youth interns developed TikTok-style messages focused on strengthening control beliefs and promoting a youth-selected mental health support resource. Youth interns distributed an online survey experiment to test whether youth-created messages shown alongside resource information increased help-seeking self-efficacy compared to an information-only control. The YPACR framework contributed to youth-relevant campaign goals, study measurements, recruitment approaches, data interpretation, and message design. We discuss the benefits and challenges of this youth-driven health campaign development model and recommendations for future research.

The role of the AP-1 adaptor complex in outgoing and incoming membrane traffic.

The AP-1 adaptor complex is found in all eukaryotes, but it has been implicated in different pathways in different organisms. To look directly at AP-1 function, we generated stably transduced HeLa cells coexpressing tagged AP-1 and various tagged membrane proteins. Live cell imaging showed that AP-1 is recruited onto tubular carriers trafficking from the Golgi apparatus to the plasma membrane, as well as onto transferrin-containing early/recycling endosomes. Analysis of single AP-1 vesicles showed that they are a heterogeneous population, which starts to sequester cargo 30 min after exit from the ER. Vesicle capture showed that AP-1 vesicles contain transmembrane proteins found at the TGN and early/recycling endosomes, as well as lysosomal hydrolases, but very little of the anterograde adaptor GGA2. Together, our results support a model in which AP-1 retrieves proteins from post-Golgi compartments back to the TGN, analogous to COPI's role in the early secretory pathway. We propose that this is the function of AP-1 in all eukaryotes.

The One Hour Human Proteome.

We describe deep analysis of the human proteome in less than 1 h. We achieve this expedited proteome characterization by leveraging state-of-the-art sample preparation, chromatographic separations, and data analysis tools, and by using the new Orbitrap Astral mass spectrometer equipped with a quadrupole mass filter, a high-field Orbitrap mass analyzer, and an asymmetric track lossless (Astral) mass analyzer. The system offers high tandem mass spectrometry acquisition speed of 200 Hz and detects hundreds of peptide sequences per second within data-independent acquisition or data-dependent acquisition modes of operation. The fast-switching capabilities of the new quadrupole complement the sensitivity and fast ion scanning of the Astral analyzer to enable narrow-bin data-independent analysis methods. Over a 30-min active chromatographic method consuming a total analysis time of 56 min, the Q-Orbitrap-Astral hybrid MS collects an average of 4319 MS1 scans and 438,062 tandem mass spectrometry scans per run, producing 235,916 peptide sequences (1% false discovery rate). On average, each 30-min analysis achieved detection of 10,411 protein groups (1% false discovery rate). We conclude, with these results and alongside other recent reports, that the 1-h human proteome is within reach.

Having a Voice: Resident Perceptions of Supervision, Decision-Making and Patient Care Ownership.

OBJECTIVE: Heightened resident supervision due to patient safety concerns is increasingly common in pediatrics and may leave residents with fewer opportunities for independent decision-making, a diminished sense of autonomy, and decreased engagement. This may ultimately threaten their development into competent clinicians. Understanding how pediatric residents experience supervision's influence on their involvement in decision-making, engagement in patient care, and learning is crucial to safeguard their transition to independent practice. In relation to supervision, our research investigated: 1) how residents navigated their involvement with clinical decision-making and 2) how opportunities to make clinical decisions influenced their engagement in patient care and learning. METHODS: From 2019-2020, we recruited 38 pediatric residents from three different programs for a qualitative interview-based study. Through a constructivist stance, we explored clinical decision-making experiences and performed thematic analysis using an iterative and inductive process. RESULTS: We identified three themes: 1) Residents perceived having autonomy when they had space to make independent decisions, regardless of supervisor's presence; 2) Patient care ownership resulted from having a voice in a variety of contributions to patient care; and 3) Supervisors' behaviors modulated patient care ownership and thereby residents' sense of feeling heard, their engagement in patient care, and their learning. CONCLUSIONS: Our results suggest that focusing on patient care ownership may better fit with current learning environments than aiming for independence and autonomy. They provide insight on how, in the pediatric learning climate of enhanced supervision, supervisors can preserve resident engagement in patient care and learning by augmenting patient care ownership and ensuring residents have a voice.

Fast and Deep Phosphoproteome Analysis with the Orbitrap Astral Mass Spectrometer.

Owing to its roles in cellular signal transduction, protein phosphorylation plays critical roles in myriad cell processes. That said, detecting and quantifying protein phosphorylation has remained a challenge. We describe the use of a novel mass spectrometer (Orbitrap Astral) coupled with data-independent acquisition (DIA) to achieve rapid and deep analysis of human and mouse phosphoproteomes. With this method we map approximately 30,000 unique human phosphorylation sites within a half-hour of data collection. We applied this approach to generate a phosphoproteome multi-tissue atlas of the mouse. Altogether, we detected 81,120 unique phosphorylation sites within 12 hours of measurement. With this unique dataset, we examine the sequence and structural context of protein phosphorylation. Finally, we highlight the discovery potential of this resource with multiple examples of novel phosphorylation events relevant to mitochondrial and brain biology.

What Residents Want: Perceptions of Learning During a Pediatrics Night Float Rotation.

OBJECTIVE: Most efforts to improve the educational value of night shifts focus on delivering content through structured sessions. Less is known about aligning curricular efforts with inherent nighttime learning. This study explored interns' nighttime experiences to better understand how learning works for the purpose of designing a curriculum to best support interns' learning at night. METHODS: The authors employed a constructivist grounded theory approach. They conducted semistructured interviews with 12 Family Medicine and Pediatric interns recruited during their first-night float rotation at a tertiary care children's hospital between February 2020 and August 2021. Interviews elicited stories about nighttime experiences on the basis of a modified critical incident technique. Four authors used an inductive approach to data analysis and codebook development, then all authors participated in a thematic review. RESULTS: The authors identified distinctions between interns' perceptions of teaching and learning, with participants reporting rich instances of experiential learning at night. The authors discovered that interns do not want a didactic teaching curriculum at night. Rather, they want support to optimize workplace learning: the opportunity to independently initiate patient assessments, informal teaching arising from patient care, reassurance that support from supervisors is readily available, orientation to resources, and feedback. CONCLUSIONS: Findings suggest informal workplace learning is already occurring at night and historical attempts to implement formal curricula may have a low return on investment. A curricular frameshift is recommended to support learning at night that emphasizes informal teaching responsive to learning needs that arise from patient care, integrating but not emphasizing formal didactics when necessary.

Stable endocytic structures navigate the complex pellicle of apicomplexan parasites.

Apicomplexan parasites have immense impacts on humanity, but their basic cellular processes are often poorly understood. Where endocytosis occurs in these cells, how conserved this process is with other eukaryotes, and what the functions of endocytosis are across this phylum are major unanswered questions. Using the apicomplexan model Toxoplasma, we identified the molecular composition and behavior of unusual, fixed endocytic structures. Here, stable complexes of endocytic proteins differ markedly from the dynamic assembly/disassembly of these machineries in other eukaryotes. We identify that these endocytic structures correspond to the 'micropore' that has been observed throughout the Apicomplexa. Moreover, conserved molecular adaptation of this structure is seen in apicomplexans including the kelch-domain protein K13 that is central to malarial drug-resistance. We determine that a dominant function of endocytosis in Toxoplasma is plasma membrane homeostasis, rather than parasite nutrition, and that these specialized endocytic structures originated early in infrakingdom Alveolata likely in response to the complex cell pellicle that defines this medically and ecologically important ancient eukaryotic lineage.

Overpoliced and Underrepresented: Perspectives on Cannabis Legalization From Members of Racialized Communities in Canada.

Historically, overpolicing of some racialized and Indigenous groups in Canada has resulted in unequal application of drug laws contributing to disproportionate rates of charges and convictions in these populations. Criminal records severely and negatively impact an individual's life and can perpetuate cycles of poverty and socioeconomic disadvantage. On October 17, 2018, Canada legalized cannabis production, distribution, sale, and possession for non-medical purposes. Advocates of criminal justice reform have raised concerns that Indigenous and racialized people may not equitably benefit from legalization due to unequal police surveillance and drug enforcement. These groups are among priority populations for research on cannabis and mental health, but their views on cannabis regulation have been largely absent from research and policy-making. To address this gap, we asked self-identified members of these communities about their lived experiences and perspectives on cannabis legalization in Canada. Between September 2018 and July 2019, we conducted semistructured interviews and focus groups with 37 individuals in Québec, Ontario, Alberta and British Columbia. During this phase of early cannabis legalization, participants responded to questions about anticipated public health risks and benefits of legalization, how their jurisdiction is responding to legalization, and what community resources would be needed to address legalization impacts. We conducted a thematic analysis and identified five major themes in the data related to race and early cannabis legalization: overpolicing of racialized communities, severity of penalties in new cannabis legislation, increased police powers, and underrepresentation of racialized groups in the legal cannabis market and in cannabis research. Participants discussed opportunities to support cannabis justice, including establishing priority licenses, issuing pardons or expunging criminal records, and reinvesting cannabis revenue into impacted communities. This work begins to address the paucity of Indigenous and racialized voices in cannabis research and identifies potential solutions to injustices of cannabis prohibition.

The impact of a stress management intervention including cultural components on stress biomarker levels and mental health indicators among indigenous women.

We examined the effectiveness of a 26-week culture-inclusive intervention on reducing salivary stress biomarker levels, and perceived stress, depressive, and post-traumatic stress disorder (PTSD) symptoms measured using scales in 53 Indigenous women in Ontario, Canada. Statistical analyses compared the average biomarker levels, and the area under the curve (AUC) of biomarkers. Differences in biomarkers and mental health scale scores pre- and post-intervention were compared using mixed models with a random intercept. Interaction terms were included between the intervention and age, education, disability, and HIV status, individually, to test for sub-group differences. Cortisol AUC post-intervention was decreased compared to pre-intervention (ß -1.29 µg/dL; 95%CI -2.35, -0.23). There was a slight decrease in perceived stress levels (aOR: -2.80; 95%CI -5.09, -0.50). The associations were stronger among women of younger age, higher education, and no disabilities. These interventions can be effective, but future interventions should target Indigenous population sub-groups to address individual needs.

Conducting research with Indigenous Peoples in Canada: ethical and policy considerations.

The international context of Indigenous mental health and wellbeing has been shaped by a number of key works recognizing Indigenous rights. Despite international recognitions, the mental health and wellness of Indigenous Peoples continues to be negatively affected by policies that ignore Indigenous rights, that frame colonization as historical rather than ongoing, or that minimize the impact of assimilation. Research institutions have a responsibility to conduct ethical research; yet institutional guidelines, principles, and policies often serve Indigenous Peoples poorly by enveloping them into Western knowledge production. To counter epistemological domination, Indigenous Peoples assert their research sovereignty, which for the purposes of this paper we define as autonomous control over research conducted on Indigenous territory or involving Indigenous Peoples. Indigenous sovereignty might also be applied to research impacting the landscape and the web of animal and spiritual lives evoked in a phrase such as "all my relations." This narrative review of material developed in the Canadian context examines the alignment with similar work in the international context to offer suggestions and a practice-based implementation tool to support Indigenous sovereignty in research related to wellness, mental health, and substance use. The compilation of key guidelines and principles in this article is only a start; addressing deeper issues requires a research paradigm shift.

Recent insights into the mental health needs of two-spirit people.

This paper provides a review of issues impacting the mental health of two-spirit people. Research has documented high rates of poverty, disparities in rates of harassment and violence, PTSD, anxiety, depression, suicidality, and substance use. Most research has focused on mental health disparities, often framing two-spirit people as affected by intersecting oppressions (i.e., racism or colonialism intersecting with homophobia or transphobia). Most research to date has been qualitative or has drawn on small convenience samples, limiting the generalizability of findings. Large-scale quantitative data is needed to advance understanding of how mental health disparities emerge and how protective factors, such as cultural engagement, improve well-being.

Lockdown labs: Pivoting to remote learning in forensic science higher education.

Forensic Science training and education is reliant on the application of knowledge to casework scenarios and the development of key practical skills that provide a platform for career development in the field. The COVID-19 pandemic introduced a number of challenges to effectively deliver practical content online and remotely, whilst still meeting intended learning outcomes, accreditation requirements, and attaining a high level of student engagement and experience. The MSc Forensic Science programme featured in this study is a one-year degree programme with a strong emphasis on the practical elements of forensic science, and a diverse international student cohort. Therefore, the restrictions associated with the pandemic made it very difficult not only to plan the delivery of material but also to adapt the content itself for effective online and remote learning. By focusing on the intended learning outcomes, a number of innovative teaching practices were developed to successfully transition from face-to-face teaching to online and remote delivery. A range of online and practical resources were developed, including a laboratory home kit, demonstration videos, online practical technique simulations (produced by Learning Science), data analysis tasks, and interactive workshops and activities, all designed to consolidate student learning and build confidence, in preparation for such a time that on campus practical teaching could resume. The initial feedback received from these activities from both staff and students was extremely positive and the transition from classroom to online teaching was a success, as reflected in student attainment and later student feedback. Students reported that they had a better understanding of what was expected of them, including knowledge of protocols and techniques, and felt much more confident moving into the next stage of their learning development. Even though the practical laboratory sessions were the most significantly affected by the restrictions associated with the pandemic, and resulted in reduced interaction for the students, this was counteracted by virtual sessions and workshops, which gave students the opportunity to engage with each other and communicate their thoughts and opinions, ultimately building key presentation and group working skills. This case study will detail the pivot to remote learning, as well as critically evaluating the feedback from students and discussing the changes that are likely to be retained as longer-term teaching practices, versus those that were a necessary temporary addition or adjustment in response to the pandemic.

Correctional-Facility-Outbreak-Associated COVID-19 Cases Among Asymptomatic Persons Identified Through Universal Testing: Vermont, 2020.

On April 6, 2020, a confirmed COVID-19 case in a correctional facility employee (Staff A) was reported to the Vermont Department of Health (VDH). Staff A worked in the facility while symptomatic, without reporting symptoms, for 10 days. VDH and the facility conducted two facility-wide testing events, implemented symptom monitoring, and initiated contact tracing. All 197 incarcerated persons and 115 (71%) staff were tested for SARS-CoV-2; 45 (23%) incarcerated persons and 17 (10%) staff had positive results (confirmed case), of whom 37 (82%) incarcerated persons and 1 (6%) staff had asymptomatic infections. Case detection enabled isolation of incarcerated persons and staff, work exclusion of staff with COVID-19, and quarantine of staff and incarcerated persons who had close contact with persons with COVID-19. Broad-based SARS-CoV-2 testing identified more cases than symptom monitoring.

A Quantitative Intersectional Exploration of Sexual Violence and Mental Health among Bi + People: Looking within and across Race and Gender.

Young bisexual people report disparities related to mental health and sexual violence compared to their heterosexual and gay/lesbian peers. However, the majority of research in these areas does not employ an intersectional design, despite evidence that health outcomes vary by race and gender within bi + populations. The goal of this paper is to provide an intersectionally-informed exploration of the prevalence of sexual violence among a diverse sample of 112 bi + people age 18-26, as well as descriptive data on stigma, mental health, and social support. Most (82%) of participants reported at least once experience of sexual violence since the age of 16. Sexual violence was positively associated with sexual stigma, anxiety, depression, and suicidality. Nonbinary participants reported greater prevalence of violence, exposure to stigma, and worse mental health outcomes relative to cisgender participants. Nonbinary BIPOC participants reported higher levels of anxiety and depression than cisgender BIPOC participants.

A highly multiplexed quantitative phosphosite assay for biology and preclinical studies.

Reliable methods to quantify dynamic signaling changes across diverse pathways are needed to better understand the effects of disease and drug treatment in cells and tissues but are presently lacking. Here, we present SigPath, a targeted mass spectrometry (MS) assay that measures 284 phosphosites in 200 phosphoproteins of biological interest. SigPath probes a broad swath of signaling biology with high throughput and quantitative precision. We applied the assay to investigate changes in phospho-signaling in drug-treated cancer cell lines, breast cancer preclinical models, and human medulloblastoma tumors. In addition to validating previous findings, SigPath detected and quantified a large number of differentially regulated phosphosites newly associated with disease models and human tumors at baseline or with drug perturbation. Our results highlight the potential of SigPath to monitor phosphoproteomic signaling events and to nominate mechanistic hypotheses regarding oncogenesis, response, and resistance to therapy.

Poverty in Lesbian, Gay, Bisexual, Transgender, Queer, Two-Spirit, and Other Sexual and Gender Minority (LGBTQ2S+) Communities in Canada: Implications for Social Work Practice.

In this article, we draw on a recent review of the Canadian literature on poverty in lesbian, gay, bisexual, transgender, queer, two-spirit, and other sexual and gender minority (LGBTQ2S+) communities to conceptualize social work interventions that may be used to address material inequities among these groups. Our literature review, which was based on a total of 39 works, revealed distinctive expressions of poverty among younger and older LGBTQ2S+ groups, as well as racialized, newcomer, and Indigenous sexual and gender minorities. Drawing on these insights, together with theoretical frameworks grounded in intersectionality and relational poverty analysis, we conceptualize these expressions of material inequity as salient sites of social work practice and propose interventions targeting these manifestations of LGBTQ2S+ poverty at various levels. Given the centrality of anti-poverty work as part of the social work profession's commitment to social justice, and the dearth of social work literature on LGBTQ2S+ poverty, this article promises to make significant contributions to social work scholarship and professional practice.

Rag GTPases and phosphatidylinositol 3-phosphate mediate recruitment of the AP-5/SPG11/SPG15 complex.

Adaptor protein complex 5 (AP-5) and its partners, SPG11 and SPG15, are recruited onto late endosomes and lysosomes. Here we show that recruitment of AP-5/SPG11/SPG15 is enhanced in starved cells and occurs by coincidence detection, requiring both phosphatidylinositol 3-phosphate (PI3P) and Rag GTPases. PI3P binding is via the SPG15 FYVE domain, which, on its own, localizes to early endosomes. GDP-locked RagC promotes recruitment of AP-5/SPG11/SPG15, while GTP-locked RagA prevents its recruitment. Our results uncover an interplay between AP-5/SPG11/SPG15 and the mTORC1 pathway and help to explain the phenotype of AP-5/SPG11/SPG15 deficiency in patients, including the defect in autophagic lysosome reformation.