Do Not Throw Away Your Shot: Pilot Study in Improving Medical School Curricula Through Focused Vaccine Education.

Introduction: With widely prevalent vaccine hesitancy and medical mistrust, future healthcare providers require knowledge of vaccines and comfort in discussing vaccines with patients. However, many U.S. medical schools do not offer formal or elective curricula focused on vaccine education. Methods: We sought to identify the need for such a curriculum and student gaps in knowledge through a pilot study surveying first- and second-year medical students at a large Pennsylvania medical school to assess knowledge about vaccine initiatives/policies, comfort in discussing vaccines, attitudes toward curricular changes, and effects of COVID-19 on willingness for vaccine education. Results: Many participants (>40%) reported insufficient knowledge of vaccine policy, vaccine development and testing, and community vaccine initiatives. The majority (>50%) indicated discomfort in discussing vaccines in a clinical setting. Importantly, 79% of participants reported insufficient coverage of vaccine topics in the current curriculum. A total of 54% decisively wanted formal/mandatory education versus 72% who decisively wanted elective education. Desiring formal education was associated with less comfort in discussing vaccines overall and with vaccine-hesitant people in clinical settings. Reassuringly, 74% of participants noted that the COVID-19 pandemic made them want to learn more about vaccines. Guided by these findings, we conducted a pilot elective teaching about vaccines through case studies, interactive discussions, and role-playing scenarios. Pre- and postcourse surveys showed improvement in student knowledge in all vaccine-related topics assessed. Conclusions: These data can inform the development of formal and elective curricula that effectively prepare medical students for patients' vaccine questions.

A Cross-Sectional Factor Analysis of COVID-19 and Influenza Vaccination Decisions in a Racially Diverse Western Pennsylvania Community.

INTRODUCTION: Influenza and COVID-19 vaccination rates remain suboptimal, demanding new community-centric approaches that improve targeted counseling and increase vaccine uptake. Notably, racially diverse communities show high vaccine hesitancy, yet most existing vaccine studies focus on white, college-educated cohorts. OBJECTIVE: Here, we identify factors influencing vaccination decisions of patients at Turtle Creek Primary Care clinic in Turtle Creek, PA, a racially-diverse borough. DESIGN: A retrospective mixed-methods study of the predominantly non-white patient population at Turtle Creek Primary Care clinic, a clinic caring for >70% minority patients. RESULTS: Fourteen factors emerged that patients reported were crucial to vaccine decision-making. Of these factors, top reasons for COVID-19 vaccine hesitancy were trust in vaccines, vaccine side effects, perceived vaccine knowledge, and faith/religion. Top reasons for influenza vaccine hesitancy were perceived need, vaccine side effects, trust in vaccines, and habitual behaviors. We also uncovered correlations between vaccine decision factors and sociodemographic factors. Participants > 65-years-old were more likely to cite personal safety in choosing to get the COVID-19 vaccine, while non-white participants were more likely to cite others' safety. Participants > 65-years-old were also more likely to cite personal safety in influenza vaccine decision-making, and non-female participants were more likely to cite perceived need for influenza vaccines. CONCLUSIONS: These data uncover targetable factors contributing to vaccine hesitancy and aid in developing community-centered, personalized vaccine education approaches in Turtle Creek and analogous minority communities.

A Cross-Sectional Analysis of Community Perceptions of Flu and COVID-19 Vaccines at Turtle Creek Primary Care Center.

BACKGROUND: Influenza (flu) and COVID-19 vaccination rates are subpar across the US, especially in racial and/or socioeconomic minority groups who are understudied in public health literature. OBJECTIVE: The objective of this mixed-methods study was to elucidate attitudes of patients at the Turtle Creek Primary Care Center, a clinic that cares for ∼70% non-white patients, towards flu and COVID-19 vaccines, with the goal of establishing vaccine education gaps and increasing vaccine uptake in minority communities. DESIGN/PATIENTS: This study was conducted as a cross-sectional analysis. Authors completed 123 patient phone surveys of patients cared for at the Turtle Creek clinic inquiring about flu and COVID-19 infection status and vaccination uptake (August 26-October 10, 2021). APPROACH/KEY RESULTS: We found that rates of vaccination were subpar in the Turtle Creek community, with only 54% having received the COVID-19 vaccine and only 44% receiving the flu vaccine regularly. There was a strong association between COVID-19 and flu vaccine acceptance and a notable correlation between vaccine acceptance and age. When assessing how vaccine acceptance was influenced by trusted sources of information, those who cited trusting "medical professionals" and "word of mouth" had higher odds of COVID-19 vaccine acceptance but those who cited trusting "social media" had decreased odds of acceptance. Finally, we uncovered 14 common factors for either vaccine acceptance or refusal that clustered into four overarching themes of trust, need, safety, and availability. CONCLUSION: These data highlight the necessity of improved vaccine education and reveal targetable populations and approaches for disseminating vaccine information.

Reduced hepatocyte mitophagy is an early feature of NAFLD pathogenesis and hastens the onset of steatosis, inflammation and fibrosis.

Nonalcoholic fatty liver disease (NAFLD) encompasses a spectrum of pathologies that includes steatosis, steatohepatitis (NASH) and fibrosis and is strongly associated with insulin resistance and type 2 diabetes. Changes in mitochondrial function are implicated in the pathogenesis of NAFLD, particularly in the transition from steatosis to NASH. Mitophagy is a mitochondrial quality control mechanism that allows for the selective removal of damaged mitochondria from the cell via the autophagy pathway. While past work demonstrated a negative association between liver fat content and rates of mitophagy, when changes in mitophagy occur during the pathogenesis of NAFLD and whether such changes contribute to the primary endpoints associated with the disease are currently poorly defined. We therefore undertook the studies described here to establish when alterations in mitophagy occur during the pathogenesis of NAFLD, as well as to determine the effects of genetic inhibition of mitophagy via conditional deletion of a key mitophagy regulator, PARKIN, on the development of steatosis, insulin resistance, inflammation and fibrosis. We find that loss of mitophagy occurs early in the pathogenesis of NAFLD and that loss of PARKIN hastens the onset but not severity of key NAFLD disease features. These observations suggest that loss of mitochondrial quality control in response to nutritional stress may contribute to mitochondrial dysfunction and the pathogenesis of NAFLD.

Hepatocyte-derived GDF15 suppresses feeding and improves insulin sensitivity in obese mice.

Growth differentiation factor 15 (GDF15) is a stress-induced secreted protein whose circulating levels are increased in the context of obesity. Recombinant GDF15 reduces body weight and improves glycemia in obese models, which is largely attributed to the central action of GDF15 to suppress feeding and reduce body weight. Despite these advances in knowledge, the tissue-specific sites of GDF15 production during obesity are unknown, and the effects of modulating circulating GDF15 levels on insulin sensitivity have not been evaluated directly. Here, we demonstrate that hepatocyte Gdf15 expression is sufficient for changes in circulating levels of GDF15 during obesity and that restoring Gdf15 expression specifically in hepatocytes of Gdf15 knockout mice results in marked improvements in hyperinsulinemia, hepatic insulin sensitivity, and to a lesser extent peripheral insulin sensitivity. These data support that liver hepatocytes are the primary source of circulating GDF15 in obesity.

Implementation and challenges of portable blood gas measurements in air medical transport.

OBJECTIVES: Ventilator management in prehospital settings using end-tidal CO2 can lead to inappropriate ventilation in the absence of point of care blood gas (POCBG) measurements. Implementation of POCBG testing in helicopter Emergency Medical Services (HEMS) is limited in part because of concern for preanalytical and analytical errors due to altitude, vibration, and other associated environmental factors and due to insufficient documentation of implementation challenges. METHODS: We performed accuracy and precision verification studies using standard materials tested pre-, in-, and post-flight (n=10) in a large HEMS agency. Quality assurance error log data were extracted and summarized for common POCBG errors during the first 31 months of use and air medical transport personnel were surveyed regarding POCBG use (n=63). RESULTS: No clinically significant differences were found between pre-, in-, and post-flight blood gas measurements. Error log data demonstrated a reduction in device errors over time. Survey participants found troubleshooting device errors and learning new clinical processes to be the largest barriers to implementation. Continued challenges for participants coincided with error log data including temperature and sampling difficulties. Survey participants indicated that POCBG testing improved patient management. CONCLUSIONS: POCBG testing does not appear to be compromised by the HEMS environment. Temperature excursions can be reduced by use of insulated transport bags with heating and cooling packs. Availability of POCBG results in air medical transport appeared to improve ventilator management, increase recognition of ventilation-perfusion mismatch, and improve patient tolerance of ventilation.

Liver-specific Prkn knockout mice are more susceptible to diet-induced hepatic steatosis and insulin resistance.

OBJECTIVE: PARKIN is an E3 ubiquitin ligase that regulates mitochondrial quality control through a process called mitophagy. Recent human and rodent studies suggest that loss of hepatic mitophagy may occur during the pathogenesis of obesity-associated fatty liver and contribute to changes in mitochondrial metabolism associated with this disease. Whole-body Prkn knockout mice are paradoxically protected against diet-induced hepatic steatosis; however, liver-specific effects of Prkn deficiency cannot be discerned in this model due to pleotropic effects of germline Prkn deletion on energy balance and subsequent protection against diet-induced obesity. We therefore generated the first liver-specific Prkn knockout mouse strain (LKO) to directly address the role of hepatic Prkn. METHODS: Littermate control (WT) and LKO mice were fed regular chow (RC) or high-fat diet (HFD) and changes in body weight and composition were measured over time. Liver mitochondrial content was assessed using multiple, complementary techniques, and mitochondrial respiratory capacity was assessed using Oroboros O2K platform. Liver fat was measured biochemically and assessed histologically, while global changes in hepatic gene expression were measured by RNA-seq. Whole-body and tissue-specific insulin resistance were assessed by hyperinsulinemic-euglycemic clamp with isotopic tracers. RESULTS: Liver-specific deletion of Prkn had no effect on body weight or adiposity during RC or HFD feeding; however, hepatic steatosis was increased by 45% in HFD-fed LKO compared with WT mice (P < 0.05). While there were no differences in mitochondrial content between genotypes on either diet, mitochondrial respiratory capacity and efficiency in the liver were significantly reduced in LKO mice. Gene enrichment analyses from liver RNA-seq results suggested significant changes in pathways related to lipid metabolism and fibrosis in HFD-fed Prkn knockout mice. Finally, whole-body insulin sensitivity was reduced by 35% in HFD-fed LKO mice (P < 0.05), which was primarily due to increased hepatic insulin resistance (60% of whole-body effect; P = 0.11). CONCLUSIONS: These data demonstrate that PARKIN contributes to mitochondrial homeostasis in the liver and plays a protective role against the pathogenesis of hepatic steatosis and insulin resistance.

Prenatal hypoxia increases susceptibility to kidney injury.

Prenatal hypoxia is a gestational stressor that can result in developmental abnormalities or physiological reprogramming, and often decreases cellular capacity against secondary stress. When a developing fetus is exposed to hypoxia, blood flow is preferentially redirected to vital organs including the brain and heart over other organs including the kidney. Hypoxia-induced injury can lead to structural malformations in the kidney; however, even in the absence of structural lesions, hypoxia can physiologically reprogram the kidney leading to decreased function or increased susceptibility to injury. Our investigation in mice reveals that while prenatal hypoxia does not affect normal development of the kidneys, it primes the kidneys to have an increased susceptibility to kidney injury later in life. We found that our model does not develop structural abnormalities when prenatally exposed to modest 12% O2 as evident by normal histological characterization and gene expression analysis. Further, adult renal structure and function is comparable to mice exposed to ambient oxygen throughout nephrogenesis. However, after induction of kidney injury with a nephrotoxin (cisplatin), the offspring of mice housed in hypoxia exhibit significantly reduced renal function and proximal tubule damage following injury. We conclude that exposure to prenatal hypoxia in utero physiologically reprograms the kidneys leading to increased susceptibility to injury later in life.

Von Hippel-Lindau Acts as a Metabolic Switch Controlling Nephron Progenitor Differentiation.

BACKGROUND: Nephron progenitors, the cell population that give rise to the functional unit of the kidney, are metabolically active and self-renew under glycolytic conditions. A switch from glycolysis to mitochondrial respiration drives these cells toward differentiation, but the mechanisms that control this switch are poorly defined. Studies have demonstrated that kidney formation is highly dependent on oxygen concentration, which is largely regulated by von Hippel-Lindau (VHL; a protein component of a ubiquitin ligase complex) and hypoxia-inducible factors (a family of transcription factors activated by hypoxia). METHODS: To explore VHL as a regulator defining nephron progenitor self-renewal versus differentiation, we bred Six2-TGCtg mice with VHLlox/lox mice to generate mice with a conditional deletion of VHL from Six2+ nephron progenitors. We used histologic, immunofluorescence, RNA sequencing, and metabolic assays to characterize kidneys from these mice and controls during development and up to postnatal day 21. RESULTS: By embryonic day 15.5, kidneys of nephron progenitor cell-specific VHL knockout mice begin to exhibit reduced maturation of nephron progenitors. Compared with controls, VHL knockout kidneys are smaller and developmentally delayed by postnatal day 1, and have about half the number of glomeruli at postnatal day 21. VHL knockout nephron progenitors also exhibit persistent Six2 and Wt1 expression, as well as decreased mitochondrial respiration and prolonged reliance on glycolysis. CONCLUSIONS: Our findings identify a novel role for VHL in mediating nephron progenitor differentiation through metabolic regulation, and suggest that VHL is required for normal kidney development.