Juneteenth in STEMM and the barriers to equitable science.

We are 52 Black scientists. Here, we establish the context of Juneteenth in STEMM and discuss the barriers Black scientists face, the struggles they endure, and the lack of recognition they receive. We review racism's history in science and provide institutional-level solutions to reduce the burdens on Black scientists.

Learning from HBCUs: How to produce Black professionals in STEMM.

Historically Black colleges and universities (HBCUs) offer high-quality education and produce leaders from various backgrounds, mainly being African American. Predominately White institutions can utilize practices that make HBCUs successful to mentor and graduate students of all backgrounds. We also suggest ways to bolster HBCUs so they can train more students.

Time management for STEMM students during the continuing pandemic.

One of the biggest obstacles to success is a lack of practical time management skills. Here, we provide suggestions on how to optimize time management.

Three prime repair exonuclease 1 preferentially degrades the integration-incompetent HIV-1 DNA through favorable kinetics, thermodynamic, structural, and conformational properties.

HIV-1 integration into the human genome is dependent on 3'-processing of the viral DNA. Recently, we reported that the cellular Three Prime Repair Exonuclease 1 (TREX1) enhances HIV-1 integration by degrading the unprocessed viral DNA, while the integration-competent 3'-processed DNA remained resistant. Here, we describe the mechanism by which the 3'-processed HIV-1 DNA resists TREX1-mediated degradation. Our kinetic studies revealed that the rate of cleavage (kcat) of the 3'-processed DNA was significantly lower (approximately 2-2.5-fold) than the unprocessed HIV-1 DNA by TREX1. The kcat values of human TREX1 for the processed U5 and U3 DNA substrates were 3.8 s-1 and 4.5 s-1, respectively. In contrast, the unprocessed U5 and U3 substrates were cleaved at 10.2 s-1 and 9.8 s-1, respectively. The efficiency of degradation (kcat/Km) of the 3'-processed DNA (U5-70.2 and U3-28.05 pM-1s-1) was also significantly lower than the unprocessed DNA (U5-103.1 and U3-65.3 pM-1s-1). Furthermore, the binding affinity (Kd) of TREX1 was markedly lower (∼2-fold) for the 3'-processed DNA than the unprocessed DNA. Molecular docking and dynamics studies revealed distinct conformational binding modes of TREX1 with the 3'-processed and unprocessed HIV-1 DNA. Particularly, the unprocessed DNA was favorably positioned in the active site with polar interactions with the catalytic residues of TREX1. Additionally, a stable complex was formed between TREX1 and the unprocessed DNA compared the 3'-processed DNA. These results pinpoint the mechanism by which TREX1 preferentially degrades the integration-incompetent HIV-1 DNA and reveal the unique structural and conformational properties of the integration-competent 3'-processed HIV-1 DNA.

Unique Interactions of the Small Translocases of the Mitochondrial Inner Membrane (Tims) in Trypanosoma brucei.

The infectious agent for African trypanosomiasis, Trypanosoma brucei, possesses a unique and essential translocase of the mitochondrial inner membrane, known as the TbTIM17 complex. TbTim17 associates with six small TbTims (TbTim9, TbTim10, TbTim11, TbTim12, TbTim13, and TbTim8/13). However, the interaction patterns of these smaller TbTims with each other and TbTim17 are not clear. Through yeast two-hybrid (Y2H) and co-immunoprecipitation analyses, we demonstrate that all six small TbTims interact with each other. Stronger interactions were found among TbTim8/13, TbTim9, and TbTim10. However, TbTim10 shows weaker associations with TbTim13, which has a stronger connection with TbTim17. Each of the small TbTims also interacts strongly with the C-terminal region of TbTim17. RNAi studies indicated that among all small TbTims, TbTim13 is most crucial for maintaining the steady-state levels of the TbTIM17 complex. Further analysis of the small TbTim complexes by size exclusion chromatography revealed that each small TbTim, except for TbTim13, is present in ~70 kDa complexes, possibly existing in heterohexameric forms. In contrast, TbTim13 is primarily present in the larger complex (>800 kDa) and co-fractionates with TbTim17. Altogether, our results demonstrate that, relative to other eukaryotes, the architecture and function of the small TbTim complexes are specific to T. brucei.

Understanding Physical Distancing and Face Mask Use Across High-Risk African American Subgroups During the COVID-19 Pandemic: Application of Health Belief Model.

Physical distancing and face masks remain frontline prevention strategies due to suboptimal vaccine uptake and the highly infectious COVID-19 variants. Communities of color are disproportionately impacted by a chronic disease burden that places them at higher risk of severe COVID-19 disease. Therefore, they can greatly benefit from face mask use and physical distancing, especially if the individual(s) have not received the vaccine. We applied the Health Belief Model to explore barriers and motivators influencing physical distancing and face mask use among high-risk, Black American subgroups during the early COVID-19 pandemic stages. We conducted 62 semi-structured interviews among four Black American subgroups: young adults, individuals with underlying medical conditions, essential workers, and parents. Thematic analysis, guided by the Health Belief Model, yielded six themes: (1) Knowledge on Face Mask Use and Physical Distancing, (2) Perceived Susceptibility and Severity Varies by Subgroup, (3) Experience with and Perceived Self-Efficacy to Engage in Preventive Behavior, (4) Perceived Benefits to engaging in preventive behaviors, (5) Perceived Barriers to engage in preventive behaviors, and (6) Cues to action to increase participation. Each subgroup's unique experience informed multilevel, tailored approaches that can be used by health promotion practitioners to improve face mask use and physical distancing among uniquely vulnerable Black American subgroups in the current and future pandemic.

How to be a GREAT mentor.

Formal training in how to mentor is not generally available to students, postdoctoral fellows, or junior faculty. We provide here a framework to develop as a mentor, using the GREAT model. This includes giving opportunities and opening doors; reaching out to help students identify their strengths and reach their goals; encouraging them by serving as a positive example; advising each mentee as an individual; and training them for independent thinking. In this personal view, we expand on each of these steps to illustrate how to develop a personalized mentoring style of your own. By combining these approaches, you as a mentor can work with your mentees to develop an effective and productive mentoring relationship.NEW & NOTEWORTHY We provide here a framework to develop as a mentor, using the GREAT model. This includes giving opportunities and opening doors; reaching out to help students identify their strengths and reach their goals; encouraging them by serving as a positive example; advising each mentee as an individual; and training them for independent thinking.

Use of Community Listening Sessions to Disseminate Research Findings to Past Participants and Communities.

The purpose of this sequential, explanatory mixed methods study is to determine changes in attitudes towards research, trust in medical researchers and the process, and willingness to participate in research among African Americans immediately after receiving past study findings in a community listening session (CLS). We developed and implemented four CLSs with a total of 57 African Americans who were either past research participants or members of the community-at-large. In the quantitative (dominant) phase, 32 participants completed pre-post surveys and 10 of those participants completed the follow-up semi-structured interviews. Paired samples t-tests and McNemar's test determined bivariate differences between pre- and post-surveys. Thematic analyses determined emerging themes to further understand these differences. There was a significant increase in: (1) perceived advantages of clinical trials pretest (M = 26.63, SD = 5.43) and post-test (M = 28.53, SD = 4.24, p < .01); and (2) in trust in medical researchers from pre to post (M = 36.16, SD = 10.40 vs. M = 27.53, SD = 9.37, p < 0.001). There was no significant difference in pre- and post-tests as it relates to perceived disadvantages of clinical trials and willingness to participate. Qualitative analysis yielded the following themes: (1) sharing research results and the impact on attitudes towards research; (2) community listening sessions: a trust building strategy; and (3) satisfaction with the community listening session. Community listening sessions hold promise as a method that researchers can use to simultaneously disseminate research findings and positively impact research perceptions and potentially participation among racial and ethnic minorities.

Arkadia (RING Finger Protein 111) Mediates Sumoylation-Dependent Stabilization of Nrf2 Through K48-Linked Ubiquitination.

BACKGROUND/AIMS: The transcription factor Nrf2 is a master regulator of the antioxidant defense system, protecting cells from oxidative damage. We previously reported that the SUMO-targeted E3 ubiquitin ligase (STUbL), RING finger protein 4 (RNF4) accelerated the degradation rate of Nrf2 in promyelocytic leukemia-nuclear body (PML-NB)-enriched fractions and decreased Nrf2-mediated gene transcription. The mechanisms that regulate Nrf2 nuclear levels are poorly understood. In this study, we aim to explore the role of the second mammalian STUbL, Arkadia/RNF111 on Nrf2. METHODS: Arkadia mediated ubiquitination was detected using co-immunoprecipitation assays in which whole cell lysates were immunoprecipated with anti-Nrf2 antibody and Western blotted with anti-hemagglutinin (HA) antibody or anti-Lys-48 ubiquitin-specific antibody. The half-life of Nrf2 was detected in whole cell lysates and promyelocytic leukemia-nuclear body enriched fractions by cycloheximide-chase. Reporter gene assays were performed using the antioxidant response element (ARE)-containing promoter Heme oxygenase-1 (HO-1). RESULTS: We show that Arkadia/RNF111 is able to ubiquitinate Nrf2 resulting in the stabilization of Nrf2. This stabilization was mediated through Lys-48 ubiquitin chains, contrary to traditionally degradative role of Lys-48 ubiquitination, suggesting that Lys-48 ubiquitination of Nrf2 protects Nrf2 from degradation thereby allowing Nrf2-dependent gene transcription. CONCLUSION: Collectively, these findings highlight a novel mechanism to positively regulate nuclear Nrf2 levels in response to oxidative stress through Arkadia-mediated K48-linked ubiquitination of Nrf2.

Structural insights into interactions of C/EBP transcriptional activators with the Taz2 domain of p300.

Members of the C/EBP family of transcription factors bind to the Taz2 domain of p300/CBP and mediate its phosphorylation through the recruitment of specific kinases. Short sequence motifs termed homology boxes A and B, which comprise their minimal transactivation domains (TADs), are conserved between C/EBP activators and are necessary for specific p300/CBP binding. A possible mode of interaction between C/EBP TADs and the p300 Taz2 domain was implied by the crystal structure of a chimeric protein composed of residues 1723-1818 of p300 Taz2 and residues 37-61 of C/EBPℇ. The segment corresponding to the C/EBPℇ TAD forms two orthogonally disposed helices connected by a short linker and interacts with the core structure of Taz2 from a symmetry-related molecule. It is proposed that other members of the C/EBP family interact with the Taz2 domain in the same manner. The position of the C/EBPℇ peptide on the Taz2 protein interaction surface suggests that the N-termini of C/EBP proteins are unbound in the C/EBP-p300 Taz2 complex. This observation is in agreement with the known location of the docking site of protein kinase HIPK2 in the C/EBPß N-terminus, which associates with the C/EBPß-p300 complex.

Three Prime Repair Exonuclease 1 preferentially degrades the integration-incompetent HIV-1 DNA through favorable kinetics, thermodynamic, structural and conformational properties.

HIV-1 integration into the human genome is dependent on 3'-processing of the reverse transcribed viral DNA. Recently, we reported that the cellular Three Prime Repair Exonuclease 1 (TREX1) enhances HIV-1 integration by degrading the unprocessed viral DNA, while the integration-competent 3'-processed DNA remained resistant. Here, we describe the mechanism by which the 3'-processed HIV-1 DNA resists TREX1-mediated degradation. Our kinetic studies revealed that the rate of cleavage (kcat) of the 3'-processed DNA was significantly lower than the unprocessed HIV-1 DNA by TREX1. The efficiency of degradation (kcat/KM) of the 3'-processed DNA was also significantly lower than the unprocessed DNA. Furthermore, the binding affinity (Kd) of TREX1 was markedly lower to the 3'-processed DNA compared to the unprocessed DNA. Molecular docking and dynamics studies revealed distinct conformational binding modes of TREX1 with the 3'-processed and unprocessed HIV-1 DNA. Particularly, the unprocessed DNA was favorably positioned in the active site with polar interactions with the catalytic residues of TREX1. Additionally, a stable complex was formed between TREX1 and the unprocessed DNA compared the 3'-processed DNA. These results pinpoint the biochemical mechanism by which TREX1 preferentially degrades the integration-incompetent HIV-1 DNA and reveal the unique structural and conformational properties of the integration-competent 3'-processed HIV-1 DNA.

COVID-19 risk communication gaps, needs, and strategies related to pandemic preparedness plans among vulnerable, Black American subgroups: A qualitative study.

OBJECTIVE: Improving current and future risk communication plans is critical to mitigate the COVID-19 pandemic and begin to prepare for future pandemics. Minority groups, particularly African Americans, have been limited in engagement to prepare these plans which has been demonstrated to be disadvantageous. We report findings from a qualitative study that describes gaps, needs, and strategies to improve communication among vulnerable, Black American subgroups during the COVID-19 pandemic. METHODS: Sixty-two Black Americans in uniquely, vulnerable subgroups participated in qualitative, semi-structured interviews from May to September 2020. Thematic analyses were used to identify themes. RESULTS: Participants were 16 essential workers, 16 parents, 15 young adults, and 15 individuals with underlying medical conditions. Emerging themes were: (1) Poor communication and miscommunication fueled fear and confusion; (2) Information sources and channels: How do I choose one?; (3) Communication needs were simple yet complex; (4) All information sources are not trusted information sources; (5) Preferred yet trusted channels and types of information; and (6) Dissemination of COVID Research: Why and How. Subgroups varied in information sources and processes for choosing the source, communication needs, and channels and types of information needed. They shared why they did and did not trust certain sources along with the importance of COVID research dissemination to promote informed decision-making throughout the pandemic. DISCUSSION: This study found that Black American subgroups had diverse, yet trusted and non-trusted messages, messengers, and strategies for communication and wanted research results disseminated. We describe multi-level stakeholders and strategies to help improve risk communication for pandemics, and potentially preparedness and health outcomes.

Defining Mitochondrial Cristae Morphology Changes Induced by Aging in Brown Adipose Tissue.

Mitochondria are required for energy production and even give brown adipose tissue (BAT) its characteristic color due to their high iron content and abundance. The physiological function and bioenergetic capacity of mitochondria are connected to the structure, folding, and organization of its inner-membrane cristae. During the aging process, mitochondrial dysfunction is observed, and the regulatory balance of mitochondrial dynamics is often disrupted, leading to increased mitochondrial fragmentation in aging cells. Therefore, it is hypothesized that significant morphological changes in BAT mitochondria and cristae will be present with aging. A quantitative 3D electron microscopy approach is developed to map cristae network organization in mouse BAT to test this hypothesis. Using this methodology, the 3D morphology of mitochondrial cristae is investigated in adult (3-month) and aged (2-year) murine BAT tissue via serial block face-scanning electron microscopy (SBF-SEM) and 3D reconstruction software for manual segmentation, analysis, and quantification. Upon investigation, an increase is found in mitochondrial volume, surface area, and complexity and decreased sphericity in aged BAT, alongside significant decreases in cristae volume, area, perimeter, and score. Overall, these data define the nature of the mitochondrial structure in murine BAT across aging.

Psychosocial Stressors and Coping Strategies Among African Americans During Early Stages of the COVID-19 Pandemic: a Qualitative Study.

OBJECTIVES: The disproportionate impact of coronavirus (COVID-19) on African Americans along with associated inequities in social determinants of health (SDOH) and racism increase their vulnerability to the psychosocial impact of COVID-19. This qualitative study applied the socio-ecological model (SEM) to explore psychosocial stressors, coping styles, and needs to improve psychosocial health among unique subgroups of African Americans in early pandemic stages. METHODS: Sixty-two African Americans (16 parents, 15 young adults, 16 essential workers, and 15 individuals with underlying medical conditions) participated in qualitative, semi-structured interviews between May and September 2020. Interview data were analyzed based on the SEM using thematic analysis. RESULTS: The majority (84%) reported being stressed with parents having the highest level. Four themes emerged : (1) our COVID-19 pandemic state of mind, (2) top stressors in the early stages of the COVID-19 pandemic, (3) coping strategies during COVID-19, and (4) needs during the COVID-19 pandemic to reduce stress. While there were similarities, different stressors were experienced among subgroups, which yielded different coping styles and needs from stakeholders across multi-levels to improve their psychosocial health. CONCLUSIONS: Findings suggest current and future pandemic response plans need targeted strategies across multiple levels of influence to address the psychosocial impact of the COVID-19 pandemic on African Americans.

Motivators and Barriers to COVID-19 Research Participation at the Onset of the COVID-19 Pandemic in Black Communities in the USA: an Exploratory Study.

INTRODUCTION: Black individuals in the USA continue to be underrepresented in clinical trials with low participation rates in COVID-19 research studies. Identifying participation barriers is necessary as we develop more vaccines and other treatments to address SARS-CoV-2 and associated sequelae. The purpose of this explorative, qualitative study is to apply the theory of planned behavior to understand motivators and barriers to COVID-19 research participation at the early stages of the COVID-19 pandemic. Understanding these factors is important to ultimately lead to increased vaccination rates among Black individuals, especially in strategies that increase preparedness in response to public health emergencies. METHODS: A phenomenological qualitative study design was conducted between May and September 2020 among 62 Black participants. The participants were purposefully selected from vulnerable subgroups of the Black population: essential workers, young adults, parents, and individuals with underlying medical conditions. An inductive-deductive content analysis approach was used to analyze the interview data. RESULTS: Majority (54.8%) reported willingness to participate in COVID-19 research. The following themes emerged from the interviews: (1) positivity toward research exists yet fear and distrust remain; (2) views toward COVID-19 research vary; (3) motivators to COVID-19 research participation; (4) barriers to COVID-19 research participation; and (5) potential strategies to increase COVID-19 research participation. CONCLUSIONS: Based on our findings, majority of the participants reported willingness to participate in research with observational research being the most commonly cited type of research. Providing data on the attitudes and perspectives of Black individuals and their intentions for COVID-19 research participation using TPB informs intervention targets for healthcare providers and policy makers for an equitable emergency response. Our results suggest improved communication on the research process, research opportunities, and participant testimonial through trusted sources could increase the likelihood of participation. This is especially important as we continue through the pandemic and new treatments for COVID-19 vaccines become readily available.

Catalyst for Change: Future of DEI in Academia.

In this paper, we propose ways to address diversity, equity, and inclusion (DEI) challenges and outline steps and methodologies for creating allies and empowering leaders to support DEI efforts in science, technology, engineering, mathematics, and medicine (STEMM) for underrepresented minorities (URMs).

COVID-19 Vaccination: Comparison of Attitudes, Decision-Making Processes, and Communication among Vaccinated and Unvaccinated Black Americans.

BACKGROUND: COVID-19 vaccination rates remain suboptimal among Black Americans who disproportionately experience higher hospitalization and death rates than White Americans. METHODS: We conducted a multi-method (interviews and surveys) study among 30 Black Americans (n = 16 vaccinated, n = 14 unvaccinated) to explore factors related to vaccination hesitancy, decision-making processes, and communication related to uptake. Participants were recruited by using community-driven approaches, including partner collaborations. Thematic analysis was used to analyze qualitative data, and descriptive and bivariate analysis was used for quantitative data. RESULTS: Of those unvaccinated, 79% (n = 11) stated they were delaying and 21% (n = 3) were declining vaccination indefinitely. When asked about the likelihood of vaccine initiation in 6 months and 12 months, 29% (n = 4) and 36% (n = 5), respectively, stated that they would receive the vaccine. The following themes emerged: (1) COVID-19 vaccination hesitancy exists on a continuum; (2) varied decision-making processes for COVID-19 vaccination; (3) motivators among vaccinated individuals; (4) barriers among unvaccinated individuals; (5) retrieving and navigating vaccine information within the COVID-19 infodemic; and (6) parent perspectives on child vaccination. CONCLUSIONS: Findings suggest that vaccinated and unvaccinated participants had similar and dissimilar perspectives in decision-making processes and vaccine concerns as shown in the Decision-making Processes for the COVID-19 vaccination (DePC) model. Based on these findings, future studies should further explore how factors influencing decision-making can lead to divergent outcomes for COVID-19 vaccination.

Unique interactions and functions of the mitochondrial small Tims in Trypanosoma brucei.

Trypanosoma brucei is an early divergent parasitic protozoan that causes a fatal disease, African trypanosomiasis. T. brucei possesses a unique and essential translocase of the mitochondrial inner membrane, the TbTIM17 complex. TbTim17 associates with 6 small TbTims, (TbTim9, TbTim10, TbTim11, TbTim12, TbTim13, and TbTim8/13). However, the interaction pattern of the small TbTims with each other and TbTim17 are not clear. Here, we demonstrated by yeast two-hybrid (Y2H) analysis that all six small TbTims interact with each other, but stronger interactions were found among TbTim8/13, TbTim9, and TbTim10. Each of the small TbTims also interact directly with the C-terminal region of TbTim17. RNAi studies indicated that among all small TbTims, TbTim13 is most crucial to maintain the steady-state levels of the TbTIM17 complex. Co-immunoprecipitation analyses from T. brucei mitochondrial extracts also showed that TbTim10 has a stronger association with TbTim9 and TbTim8/13, but a weaker association with TbTim13, whereas TbTim13 has a stronger connection with TbTim17. Analysis of the small TbTim complexes by size exclusion chromatography revealed that each small TbTim, except TbTim13, is present in ∼70 kDa complexes, which could be heterohexameric forms of the small TbTims. However, TbTim13 is primarily present in the larger complex (>800 kDa) and co-fractionated with TbTim17. Altogether, our results demonstrated that TbTim13 is a part of the TbTIM complex and the smaller complexes of the small TbTims likely interact with the larger complex dynamically. Therefore, relative to other eukaryotes, the architecture and function of the small TbTim complexes are specific in T. brucei .

Development of strategies for community engaged research dissemination by basic scientists: a case study.

As depicted in the translational research continuum, dissemination of research findings to past research participants and the community-at-large is integral to improving health outcomes. Blocks in translation exist in which poor dissemination is a major contributor. Limited progress has been made on how to engage basic scientists at T1 and T2 phases to meaningfully disseminate study findings to community. Our objective is to report on 4 cases of community engaged research dissemination activities among 3 basic scientists (ie, a cancer biologist, a biochemist, and a molecular biologist.): a townhall, a radio listening session, a community newsletter, and a Facebook Live segment. The Meharry Community Engagement Core dissemination team designed these activities using community informed processes. To plan and conduct these activities, a basic scientist is partnered with a community engaged researcher and a community-based organization to create a dissemination product which can be understood and potentially used by past research participants and the community-at-large. We share reflections from basic scientists, community organizations, and event participants. Finally, we provide competencies, informed by basic scientists, needed to engage in effective, community-engaged research dissemination. The activities, reflections, and competencies can be used by basic scientists and academic institutions as models to guide their community engaged research dissemination activities. This work supports the goal to bridge the translational research gap.

Protocol for isolating mice skeletal muscle myoblasts and myotubes via differential antibody validation.

Isolation of skeletal muscles allows for the exploration of many complex diseases. Here, we present a protocol for isolating mice skeletal muscle myoblasts and myotubes that have been differentiated through antibody validation. We describe steps for collecting and preparing murine skeletal tissue, myoblast cell maintenance, plating, and cell differentiation. We then detail procedures for cell incubation, immunostaining, slide preparation and storage, and imaging for immunofluorescence validation.