p97 Disease Mutations Modulate Nucleotide-Induced Conformation to Alter Protein-Protein Interactions.

The AAA+ ATPase p97/VCP adopts at least three conformations that depend on the binding of ADP and ATP and alter the orientation of the N-terminal protein-protein interaction (PPI) domain into up and down conformations. Point mutations that cause multisystem proteinopathy 1 (MSP1) are found at the interface of the N domain and D1-ATPase domain and potentially alter the conformational preferences of p97. Additionally, binding of adaptor proteins to the N-domain regulates p97s catalytic activity. We propose that p97/adaptor PPIs are coupled to p97 conformational states. We evaluated the binding of nucleotides and the adaptor proteins p37 and p47 to wild-type p97 and MSP1 mutants. Notably, p47 and p37 bind 8-fold more weakly to the ADP-bound conformation of wild-type p97 compared to the ATP-bound conformation. However, MSP1 mutants lose this nucleotide-induced conformational coupling because they destabilize the ADP-bound, down conformation of the N-domain. Loss in conformation coupling to PPIs could contribute to the mechanism of MSP1

Coenzyme Q10 deficiencies: pathways in yeast and humans.

Coenzyme Q (ubiquinone or CoQ) is an essential lipid that plays a role in mitochondrial respiratory electron transport and serves as an important antioxidant. In human and yeast cells, CoQ synthesis derives from aromatic ring precursors and the isoprene biosynthetic pathway. Saccharomyces cerevisiae coq mutants provide a powerful model for our understanding of CoQ biosynthesis. This review focusses on the biosynthesis of CoQ in yeast and the relevance of this model to CoQ biosynthesis in human cells. The COQ1-COQ11 yeast genes are required for efficient biosynthesis of yeast CoQ. Expression of human homologs of yeast COQ1-COQ10 genes restore CoQ biosynthesis in the corresponding yeast coq mutants, indicating profound functional conservation. Thus, yeast provides a simple yet effective model to investigate and define the function and possible pathology of human COQ (yeast or human gene involved in CoQ biosynthesis) gene polymorphisms and mutations. Biosynthesis of CoQ in yeast and human cells depends on high molecular mass multisubunit complexes consisting of several of the COQ gene products, as well as CoQ itself and CoQ intermediates. The CoQ synthome in yeast or Complex Q in human cells, is essential for de novo biosynthesis of CoQ. Although some human CoQ deficiencies respond to dietary supplementation with CoQ, in general the uptake and assimilation of this very hydrophobic lipid is inefficient. Simple natural products may serve as alternate ring precursors in CoQ biosynthesis in both yeast and human cells, and these compounds may act to enhance biosynthesis of CoQ or may bypass certain deficient steps in the CoQ biosynthetic pathway

Quest to Belong: Nursing Students’ Perceptions while Transitioning from College to University

Background: Students face challenges throughout their undergraduate nursing programs. Adapting to university, accessing resources, and transitioning from one year to the next and between institutions are examples of these challenges. Limited research exists on the perceptions of undergraduate nursing students’ sense of belonging while transitioning from college to university in a collaborative nursing degree program in Canada. Purpose of the Study: The purpose of the study was to examine third year undergraduate nursing students’ perception of sense of belonging while transitioning from college to university in a collaborative nursing degree program. More specifically, the study aimed to identify factors that have affected students’ experiences of their sense of belonging and identify strategies that have influenced the college site students’ sense of belonging while transitioning to the university. Methods: Interpretive Description qualitative methodology was used to explore nursing students’ perceptions of sense of belonging while transitioning from college to university. Participants were third year nursing students who had transitioned from the college site to university site. Data was collected in February 2020 by in-depth in-person semi-structured individual interviews. Thematic analysis was used to analyze the data. Results: Thirteen students participated. Four emergent themes were identified: journey of emotions, perception of barriers and challenges, facilitators to sense of belonging, and students’ suggestions for change. Conclusion: This study highlighted the factors that impacted nursing students’ sense of belonging while transitioning. Study findings revealed perceived challenges and supports from social, program, and institutional perspectives in students’ transition journey from college to university. These findings provide insights to guide faculty, collaborative programs, and academic institutions in ways to design specific strategies and guidelines to support nursing students’ success and to enhance their sense of belonging as they transition from college to university. Résumé Contexte : Les étudiantes font face à des défis tout au long du programme de premier cycle en sciences infirmières. L’adaptation à l’université, l’accès aux ressources et la transition d’une année à l’autre et entre les établissements sont des exemples de ces défis. Il existe peu de recherches sur les perceptions du sentiment d’appartenance des étudiantes en sciences infirmières de premier cycle lors de la transition du collège à l’université dans le cadre d’un programme collaboratif de grade en sciences infirmières au Canada. Objectif de l’étude : L’objectif de l’étude était d’examiner la perception du sentiment d’appartenance d’étudiantes de troisième année du premier cycle en sciences infirmières lors de la transition du collège à l’université dans le cadre d’un programme collaboratif de grade en sciences infirmières. Plus précisément, l’étude visait à déterminer les facteurs qui ont influé sur le sentiment d’appartenance des étudiantes et à cibler les stratégies qui ont influencé le sentiment d’appartenance des étudiantes du collégial lors de leur transition vers l’université. Méthodes : Une méthodologie qualitative de description interprétative a été utilisée pour explorer les perceptions du sentiment d’appartenance des étudiantes en sciences infirmières lors de la transition du collège à l’université. Les participantes étaient d’étudiantes en sciences infirmières de troisième année lors de la transition du collège à l’université. Les données ont été collectées en février 2020 par des entrevues individuelles semi-structurées approfondies en personne. Une analyse thématique a été utilisée pour analyser les données. Résultats : Treize étudiantes ont participé. Quatre thèmes émergents ont été identifiés : le cheminement des émotions, la perception des obstacles et des défis, les facilitateurs du sentiment d’appartenance et les suggestions de changement des étudiantes. Conclusion : Cette étude a mis en évidence les facteurs qui ont eu un impact sur le sentiment d’appartenance des étudiantes en sciences infirmières lors de la transition. Les résultats de l’étude ont révélé les défis et les soutiens perçus du point de vue social, des programmes et des établissements dans le parcours de transition des étudiantes du collège à l’université. Ces résultats fournissent des pistes pour guider le corps professoral, les programmes de collaboration et les établissements universitaires dans la conception de stratégies et de lignes directrices spécifiques pour soutenir la réussite des étudiantes en sciences infirmières et pour renforcer leur sentiment d’appartenance lors de leur transition du collège à l’université

Nursing Students Desire to Belong in Online Learning Environments. Part 1: A Mixed-Methods Study

Background: In early 2020, due to the declaration of the COVID-19 pandemic, many educational institutions significantly modified their learning environments from in-person classrooms to online delivery to ensure the continuity of education and limit the spread of the virus. Understanding how undergraduate nursing students experience belonging during this drastic shift to online learning is an area of inquiry that has a limited amount of research. Purpose of the Study: The goal of this study was to describe nursing students’ sense of belonging in an online learning environment as well as identify strategies and supports to foster their sense of belonging. Methods: An explanatory sequential mixed method design with two phases was used for this study. In phase one, the focus of this paper, we used a mixed-method approach. The quantitative component consisted of an online survey and the qualitative component was the three open-ended questions included as part of the survey. Participants were third-year nursing students from a collaborative nursing degree program who had recently transitioned from the college to the university site and were currently taking all courses online as a result of the COVID-19 pandemic. Results: This paper reports on phase one of the study conducted in April 2021. The survey data were analyzed using descriptive statistics and the three open-ended questions were analyzed using thematic analysis. Conclusions: As more educational institutions continue with online courses during the pandemic and post-pandemic, the findings from this study will provide valuable information as to the factors that foster or hinder nursing students’ sense of belonging in an online learning environment. These findings will also assist in contributing to the development of creative teaching modalities and pedagogies which can help to enhance the quality of student learning experiences and their sense of belonging in online learning environments. Résumé Contexte : Au début de l’année 2020, en raison de la pandémie de COVID-19, de nombreux établissements d’enseignement ont modifié considérablement leurs environnements d’apprentissage, passant de cours en présence à l’apprentissage en ligne, afin d’assurer la poursuite de la formation tout en limitant la propagation du virus. Il y a peu d’études qui permettent de comprendre comment les étudiantes et étudiants du premier cycle en sciences infirmières ont vécu le sentiment d’appartenance lors de cette transition radicale vers l’apprentissage virtuel. Buts de l’étude : Les buts de cette étude étaient de décrire le sentiment d’appartenance d’étudiantes et étudiants en sciences infirmières dans un environnement d’apprentissage en ligne ainsi que d’identifier des stratégies et des mesures de soutien qui favoriseraient leur sentiment d’appartenance. Méthodes : Un devis mixte séquentiel explicatif en deux phases a été utilisé pour cette étude. Dans la première phase, qui est au cœur de cet article, nous avons adopté une approche mixte. La composante quantitative consistait en un sondage en ligne, tandis que la composante qualitative était constituée de trois questions contenue dans le sondage. Les personnes qui ont participé à cette étude étaient des étudiantes et étudiants de troisième année en sciences infirmières inscrits à un programme collaboratif qui venaient de faire la transition du collège à l’université, puis, en raison de la pandémie de COVID-19, ils suivaient tous leurs cours en ligne. Résultats : Cet article présente les résultats de la première phase de l’étude réalisée en avril 2021. Les données recueillies par sondage ont été analysées en utilisant des statistiques descriptives, tandis que les réponses aux trois questions ouvertes ont fait l’objet d’une analyse thématique. Conclusions : Alors que de nombreux établissements d’enseignement poursuivent l’utilisation de cours en ligne au-delà de la période de pandémie, cette étude permet de comprendre les facteurs qui influencent le sentiment d’appartenance d’étudiantes et étudiants en sciences infirmières dans un contexte d’apprentissage en ligne. Les résultats obtenus contribueront également au développement de méthodes d’enseignement créatives et d’approches pédagogiques qui pourront améliorer la qualité de l’expérience d’apprentissage des étudiantes et étudiants et renforcer leur sentiment d’appartenance dans les environnements d’apprentissage en ligne

Specific Inhibition of p97/VCP ATPase and Kinetic Analysis Demonstrate Interaction between D1 and D2 ATPase domains

The p97 AAA (ATPase associated with diverse cellular activities), also called VCP (valosin-containing protein), is an important therapeutic target for cancer and neurodegenerative diseases. p97 forms a hexamer composed of two AAA domains (D1 and D2) that form two stacked rings, and an N-terminal domain that binds numerous cofactor proteins. The interplay between the three domains in p97 is complex, and a deeper biochemical understanding is needed in order to design selective p97 inhibitors as therapeutic agents. It is clear that the D2 ATPase domain hydrolyzes ATP in vitro, but whether D1 contributes to ATPase activity is controversial. Here, we use Walker A and B mutants to demonstrate that D1 is capable of hydrolyzing ATP, and show for the first time that nucleotide binding in the D2 domain increases the catalytic efficiency (k_(cat)/K_m) of D1 ATP hydrolysis 280-fold, by increasing k_(cat) 7-fold and decreasing K_m about 40-fold. We further show that an ND1 construct lacking D2 but including the linker between D1 and D2 is catalytically active, resolving a conflict in the literature. Applying enzymatic observations to small-molecule inhibitors, we show that four p97 inhibitors (DBeQ, ML240, ML241, and NMS-873) have differential responses to Walker A and B mutations, to disease-causing IBMPFD mutations, and to the presence of the N-domain binding cofactor protein p47. These differential effects provide the first evidence that p97 cofactors and disease mutations can alter p97 inhibitor potency and suggest the possibility of developing context-dependent inhibitors of p97

Specific Inhibition of p97/VCP ATPase and Kinetic Analysis Demonstrate Interaction between D1 and D2 ATPase domains

The p97 AAA (ATPase associated with diverse cellular activities), also called VCP (valosin-containing protein), is an important therapeutic target for cancer and neurodegenerative diseases. p97 forms a hexamer composed of two AAA domains (D1 and D2) that form two stacked rings, and an N-terminal domain that binds numerous cofactor proteins. The interplay between the three domains in p97 is complex, and a deeper biochemical understanding is needed in order to design selective p97 inhibitors as therapeutic agents. It is clear that the D2 ATPase domain hydrolyzes ATP in vitro, but whether D1 contributes to ATPase activity is controversial. Here, we use Walker A and B mutants to demonstrate that D1 is capable of hydrolyzing ATP, and show for the first time that nucleotide binding in the D2 domain increases the catalytic efficiency (kcat/Km) of D1 ATP hydrolysis 280-fold, by increasing kcat 7-fold and decreasing Km about 40-fold. We further show that an ND1 construct lacking D2 but including the linker between D1 and D2 is catalytically active, resolving a conflict in the literature. Applying enzymatic observations to small-molecule inhibitors, we show that four p97 inhibitors (DBeQ, ML240, ML241, and NMS-873) have differential responses to Walker A and B mutations, to disease-causing IBMPFD mutations, and to the presence of the N-domain binding cofactor protein p47. These differential effects provide the first evidence that p97 cofactors and disease mutations can alter p97 inhibitor potency and suggest the possibility of developing context-dependent inhibitors of p97

Exploring HPV vaccination policy and payer strategies for opportunities to improve uptake in safety-net settings

IntroductionWe explored priorities and perspectives on health policy and payer strategies for improving HPV vaccination rates in safety-net settings in the United States.MethodsWe conducted qualitative interviews with policy and payer representatives in the greater Los Angeles region and state of New Jersey between December 2020 and January 2022. Practice Change Model domains guided data collection, thematic analysis, and interpretation.ResultsFive themes emerged from interviews with 11 policy and 8 payer participants, including: (1) payer representatives not prioritizing HPV vaccination specifically in incentive-driven clinic metrics; (2) policy representatives noting region-specific HPV vaccine policy options; (3) inconsistent motivation across policy/payer groups to improve HPV vaccination; (4) targeting of HPV vaccination in quality improvement initiatives suggested across policy/payer groups; and (5) COVID-19 pandemic viewed as both barrier and opportunity for HPV vaccination improvement across policy/payer groups.DiscussionOur findings indicate opportunities for incorporating policy and payer perspectives into HPV vaccine improvement processes. We identified a need to translate effective policy and payer strategies, such as pay-for-performance programs, to improve HPV vaccination within safety-net settings. COVID-19 vaccination strategies and community efforts create potential policy windows for expanding HPV vaccine awareness and access

Novel Nuclear Partnering Role of EPS8 With FOXM1 in Regulating Cell Proliferation

One hallmark of cancer cells is sustaining proliferative signaling that leads to uncontrolled cell proliferation. Both the Forkhead box (FOX) M1 transcription factor and the Epidermal Growth Factor (EGF) receptor Pathway Substrate 8 (EPS8) are known to be activated by mitogenic signaling and their levels upregulated in cancer. Well-known to regulate Rac-mediated actin remodeling at the cell cortex, EPS8 carries a nuclear localization signal but its possible nuclear role remains unclear. Here, we demonstrated interaction of FOXM1 with EPS8 in yeast two-hybrid and immunoprecipitation assays. Immunostaining revealed co-localization of the two proteins during G2/M phase of the cell cycle. EPS8 became nuclear localized when CRM1/Exportin 1-dependent nuclear export was inhibited by Leptomycin B, and a functional nuclear export signal could be identified within EPS8 using EGFP-tagging and site-directed mutagenesis. Downregulation of EPS8 using shRNAs suppressed expression of FOXM1 and the FOXM1-target CCNB1, and slowed down G2/M transition in cervical cancer cells. Chromatin immunoprecipitation analysis indicated recruitment of EPS8 to the CCNB1 and CDC25B promoters. Taken together, our findings support a novel partnering role of EPS8 with FOXM1 in the regulation of cancer cell proliferation and provides interesting insight into future design of therapeutic strategy to inhibit cancer cell proliferation

Altered cofactor regulation with disease-associated p97/VCP mutations

Dominant mutations in p97/VCP (valosin-containing protein) cause a rare multisystem degenerative disease with varied phenotypes that include inclusion body myopathy, Paget’s disease of bone, frontotemporal dementia, and amyotrophic lateral sclerosis. p97 disease mutants have altered N-domain conformations, elevated ATPase activity, and altered cofactor association. We have now discovered a previously unidentified disease-relevant functional property of p97 by identifying how the cofactors p37 and p47 regulate p97 ATPase activity. We define p37 as, to our knowledge, the first known p97-activating cofactor, which enhances the catalytic efficiency (k_(cat)/K_m) of p97 by 11-fold. Whereas both p37 and p47 decrease the K_m of ATP in p97, p37 increases the k_(cat) of p97. In contrast, regulation by p47 is biphasic, with decreased k_(cat) at low levels but increased k_(cat) at higher levels. By deleting a region of p47 that lacks homology to p37 (amino acids 69–92), we changed p47 from an inhibitory cofactor to an activating cofactor, similar to p37. Our data suggest that cofactors regulate p97 ATPase activity by binding to the N domain. Induced conformation changes affect ADP/ATP binding at the D1 domain, which in turn controls ATPase cycling. Most importantly, we found that the D2 domain of disease mutants failed to be activated by p37 or p47. Our results show that cofactors play a critical role in controlling p97 ATPase activity, and suggest that lack of cofactor-regulated communication may contribute to p97-associated disease pathogenesis

The glutathione-S-transferase Mu 1 null genotype modulates ozone-induced airway inflammation in human subjects

The Glutathione-S-Transferase Mu 1 null genotype has been reported to be a risk factor for acute respiratory disease associated with increases in ambient air ozone. Ozone is known to cause an immediate decrease in lung function and increased airway inflammation. However, it is not known if GSTM1 modulates these ozone responses in vivo in human