Nutrient Utilization and Regulation in Retinal Pigment Epithelium

Purpose: The retinal pigment epithelium (RPE) plays a critical role in maintaining vision by performing essential functions including visual cycle processing, nutrient transport, protein synthesis, protection against oxidative stress, cytokine secretion, and phagocytosis of outer segments. RPE dysfunction can cause photoreceptor cell death in inherited retinal degeneration and age-related retinal degeneration (AMD), resulting in blindness. RPE relies on its robust mitochondrial metabolism to support its multifaceted functions. However, it remains unclear how healthy RPE mitochondria fuel their metabolism and how substrate utilization is impaired in diseased RPE. Nicotinamide N-methyl transferase, an enzyme that methylates nicotinamide into 1-methyl nicotinamide (1-MNAM) is a master regulator of mitochondrial metabolism. Transcriptomics data show that NNMT is highly expressed in RPE, and its expression is further upregulated in RPE from patients with age-related macular degeneration (AMD). A gap in knowledge remains regarding how NNMT regulates RPE substrate utilization and mitochondrial metabolism in RPE. The purpose of this work is to comprehensively evaluate nutrient utilization in healthy and diseased RPE and determine the role of NNMT in RPE metabolism. The central hypothesis is that healthy human RPE will have a high metabolic flexibility in nutrient utilization which will be disrupted in diseased RPE. NNMT is expressed in RPE to regulate its unique metabolism, but its excessive activity will contribute to metabolic changes in aging and AMD. Methods and Results: A novel approach was developed using Biolog Phenotype Microarray Assays to screen substrate utilization in human RPE cells. Five human RPE cells were used for this substrate screening, including dedifferentiated fetal RPE (fRPE), induced pluripotent stem cell-derived RPE (iPSC RPE), Sorsby Fundus dystrophy (SFD) patient-derived iPSC RPE, CRISPR-corrected isogenic SFD (cSFD) iPSC RPE, and ARPE-19 cell lines. Some changes were further validated with stable isotope tracing using mass spectrometry. Differentiated fRPE and healthy iPSC RPE cells could use up to 51 nutrients. However, when dedifferentiated, fRPE used far fewer nutrients, primarily sugar and glutamine-related amino acids. SFD RPE can use 37 nutrients; however, compared to cSFD RPE and healthy iPSC RPE, they were unable to use lactate, some TCA cycle intermediates, and short-chain fatty acids. Nonetheless, they showed increased use of branched-chain amino acids (BCAAs) and BCAA-containing dipeptides. Dedifferentiated ARPE-19 cells grown in traditional culture media were incapable of utilizing lactate and ketone bodies. In contrast, nicotinamide supplementation promotes differentiation toward an epithelial phenotype, restoring the ability to use these nutrients. NNMT expression was quantified by quantitative PCR and immunoblot. Human RPE cells, isolated mouse retina, and mouse RPE were incubated with deuterium nicotinamide to trace NAD synthesis and degradation. Mitochondrial oxygen consumption was measured using the Agilent Seahorse assay. Pharmacological inhibitors and constructs packaged in lentivirus or adeno-associated virus (AAV) were used for loss or gain of function studies. NNMT expression and activity were limited to the RPE but almost absent in the retina. RPE contained more 1-MNAM, the product of NNMT, than any other tissues. 1-MNAM was actively exported by the RPE to the retina. The inhibition of NNMT inhibited mitochondrial respiration and disrupted RPE nutrient utilization. Finally, NNMT was upregulated in aged RPE and RPE from AMD patients, and excessive NNMT activity impaired RPE morphology. Conclusion: Healthy RPE cells have high flexibility in using different nutrients because of their epithelial phenotype. SFD RPE cells have reduced metabolic flexibility, relying on the oxidation of BCAAs. Our findings highlight the important roles of nutrient availability and use in RPE differentiation and diseases. NNMT is enriched in RPE, and its optimal expression is crucial for RPE nutrient utilization, metabolism, and morphology. Excessive NNMT activity in RPE may underlie pathology in aging and AMD

Effects of Pubertal Stress on Later Pup-Directed Behavior in Pregnant and Non-Pregnant Adult Female Mice

Beginning in adolescence and throughout adulthood, women are at an increased risk for anxiety and mood disorders compared to men. Several risk factors for these disorders have been identified, but it is not clear how they interact. One such factor is pregnancy. The normative changes to the brain and behavior that occur during and after pregnancy are associated with an increased risk for anxiety and mood disorders. Exposure to adverse childhood experiences (ACEs) has also been associated with an increased risk of mood disorders, and compared to men, women appear to be particularly vulnerable to the lasting effects of stress that occurs during puberty. There is evidence that these factors interact and increase vulnerability to anxiety or mood disorders, but this relationship is not well understood. Previously, we found that adult female mice that had undergone chronic variable stress (CVS) in adolescence showed a blunted hypothalamic-pituitary-adrenal (HPA) axis response to acute stress in late pregnancy. In postpartum humans, this same blunted maternal stress response has been correlated with an increased incidence of depressive symptoms. However, the consequences of this HPA blunting on behavior are not well understood. Of particular interest are maternal behaviors, such as pup retrieval in mice, which emerge during pregnancy due to vast changes in hormones and neural circuitry. The current study aims to examine the effects of CVS during adolescence on pup retrieval behaviors in pregnant and not-pregnant adult female mice. We hypothesized that exposure to chronic stress during puberty would disrupt the formation of maternal behavior in adult mice. Here, we found that pubertal stress disrupted the HPA axis response of CVS mice to behavioral testing. In addition, pubertal stress led to changes in behavior on a pup retrieval task

Pinus strobus

https://researchrepository.wvu.edu/pinaceae/1551/thumbnail.jp

Haven: Viewing the Concept Album Through the Classical Lens

This research project analyzes the album Haven by Canadian pop-rock band Marianas Trench and compares it to the seminal song cycles of Franz Schubert, Winterreise and Die schöne Müllerin, as well as those of Robert Schumann, Dichterliebe and Frauenliebe und Leben. Haven is a concept album that is based on the hero’s journey framework outlined in Joseph Campbell’s The Hero With a Thousand Faces. Josh Ramsay, the lead singer of the group, crafted his hero’s journey by drawing on his own life experiences. The songs on the album represent different plot points from Campbell’s framework, telling a cohesive story over the course of the album. Ramsay utilized a variety of musical and poetic devices to tell this story, many of which can be found throughout Schubert and Schumann’s cycles as well. The comparison of these pieces shows the possibilities of utilizing popular music in university music curriculum. Haven can be utilized as a modern example of the German Romantic song cycle, providing students with music from a genre that is more relevant to modern life. It can also stand alone, exemplifying compositional practices in modern popular music and leading to a broader range of topics in curriculum

A Multi-Case Study Exploring NCAA Division I Athletic Departments’ Mental Health Culture through Stakeholders’ Perspectives

Despite the NCAA’s ongoing efforts to guide athletic departments into providing sufficient mental health support to student-athletes, psychological well-being continues to be a primary area of concern for collegiate athletes. Student-athletes are as likely as the regular college population to experience mental health issues, but less likely to seek services (Watson, 2005). Researchers have continued to examine the perceived barriers preventing student-athletes from utilization of mental health services which include a lack of knowledge, limited access, and perceived negative stigma (e.g., Harris & Maher, 2023; Yoon & Petrie, 2023). Organizational culture–loosely defined as a group’s accumulated shared learning in which adopted actions and ideologies are all designed to help increase the success of the organization (Schein & Schein, 2017)--plays a significant role in the way it binds its stakeholders and guides their behaviors. Therefore, developing a positive athletic department culture surrounding mental health can be beneficial in increasing student-athlete help-seeking behaviors. The primary purpose of this study was to investigate the employed practices of Division I athletic departments and how stakeholders perceive and experience the layers of culture (artifacts, beliefs and values, underlying assumptions; Schein & Schein, 2017) surrounding mental health. A dual-case study approach was used, and multiple sources of data were collected including semi-structured/focus group interviews (N=14 participants), archival documents, physical artifacts, and direct observation (Stake, 1995). Data was analyzed using thematic analysis (Braun & Clark, 2006) and revealed overarching themes of visibility, integration, and leadership support as imperative factors in changing athletic department culture and reducing mental health stigma. These findings highlight the most significant interacting elements of athletic department culture (Schroeder, 2010) that impact the sub-culture related to mental health. Practical recommendations for future researchers and applicable suggestions for mental health providers and athletic administrators are presented to improve athletic department culture for effective mental health support

Ex Libris, Spring 2025

The West Virginia University Libraries magazine, Ex Libris, is produced and printed through the support of library donors. AOTUS SPENDS DAY WITH WVRHC The Archivist of the United States visited WVU Morgantown for a series of visits and tours, and chatted with WVU Libraries in an exclusive Q & A interview. RISING TO MEET RESEARCH NEEDS WVU Libraries and the WVU Research Office are empowering University researchers, paving the way to advances in research and science. CENTERING STUDENT SUCCESS AMIDST CHANGE The hybrid introduction to Library Research course isn\u27t your average college course. WVU librarians are innovating to make information literacy real and relatable for students. TEN YEARS OF ART IN THE LIBRARIES From a vision to vibrant hubs of artistic expression, cultural enrichment, and visual displays of scholarship. Dive into a visual timeline of Art in the Libraries impact. FUTURE OF THE HUMANITIES CENTER The Humanities Center is embarking on a journey of evolution based on the needs of WVU\u27s humanities community. Plus, meet the new director leading the change. LIBRARIES NEWS AND UPDATES Meet the students and faculty who received awards from the Libraries, as well as the new librarians we welcomed, the current librarians we celebrated, and the retiring librarians we said farewell to. SUPPORTING THE LIBRARIES\u27 FUTURE To meet shifting needs of WVU students, researchers, and West Virginians, WVU Libraries has honed its development and giving priorities for 2025

Project FORESIGHT Annual Report, 2023-2024

Project FORESIGHT is a business-guided self-evaluation of forensic science laboratories across the globe. The participating laboratories represent local, regional, state, and national agencies. Economics, accounting, finance, and forensic faculty provide assistance, guidance, and analysis. Laboratories participating in Project FORESIGHT have developed standardized definitions for metrics to evaluate work processes, linking financial information to work tasks and functions. Laboratory managers can then assess resource allocations, efficiencies, and value of services—the mission of Project FORESIGHT is to measure, preserve what works, and change what does not. The benchmark data for the 2023-2024 performance period includes laboratory submissions for a variety of fiscal year definitions. However, all submissions have December 31, 2023 as part of their fiscal year accounting. The majority of submissions follow a July 1, 2023 through June 30, 2024 convention. Others follow a year that begins as early as January 1, 2023 (ending December 31, 2023) while the other extreme includes laboratories with a fiscal year originating October 1, 2023 and ending September 30, 2024

Soft Infrastructure and the Location Choice of Multinational Firms:Evidence from Japanese Investment in the United States in the 1980s

We examine whether culture-specific educational infrastructure influenced the location decisions of multinational enterprises by analyzing Japanese foreign direct investment (FDI) in the United States during the 1980s. Using the quasi-natural experiment of politically-driven Japanese FDI following Reagan-era trade tensions, we test whether pre-existing Japanese Studies programs in U.S. universities predict subsequent Japanese investment across 722 commuting zones. We find that zones with Japanese Studies programs in 1980 were 21-29% more likely to receive new Japanese manufacturing investment by 1992, controlling for traditional determinants likemanufacturing infrastructure, agglomeration, and market access. Our results suggest that soft infrastructure —culture-specific human capital pipelines—represents an overlooked location advantage that complements traditional hard infrastructure in attracting FDI. These findings have implications for understanding how regions can strategically develop cultural competencies to attract foreign investment

Implementation of an Evidenced Based Enteral Nutrition Protocol for Mechanically Ventilated Patients: A Quality Improvement Initiative

Abstract Implementation of an Evidenced Based Enteral Nutrition Protocol for Mechanically Ventilated Patients: A Quality Improvement Initiative. Hannah Nuzum Background: Mechanically ventilated patients require supplemental nutrition as an important factor for their overall recovery. Malnutrition in hospitalized patients is reported as high as 35% (Ikram et al., 2022). Purpose: Lack of an enteral nutrition (EN) protocol has contributed to variability in initiation of EN for post-intubated mechanically ventilated patients. Intervention: The project interventions are threefold: the development of an EN protocol, EN initiated within 24 to 28 hours for post-intubation patients, and staff understands that EN should be initiated within 24 to 48 hours after post-intubation for better patient outcomes. Methods: This change project was developed utilizing the Lewin’s Theory of Change theoretical framework to implement an EN protocol for adult intensive care unit patients. Collaboration with the information technology team will be done to integrate clinical notifications within the electronic health record, and education will be provided to staff to appropriately implement the enteral nutrition protocol into patient care. Staff will also take post intervention surveys to look at potential barriers and feasibility of the intervention. Results: EN protocol was developed and integrated within the medical intensive care unit at Ruby Memorial Hospital. The median time for initiation of EN for pre-intervention patients was 79 hours, and the median time for initiation of EN for post-intervention patients was 17 hours. Conclusions: Development of an electronic EN protocol aids in making initiation of EN for mechanically ventilated patients more consistent and improves the time to initiation of EN for critically ill patients