Genetic tuning of ß-carotene oxygenase-1 activity rescues cone photoreceptor function in STRA6-deficient mice.

Rod and cone photoreceptors in the retina mediate dim light and daylight vision, respectively. Despite their distinctive functions, rod and cone visual pigments utilize the same vitamin A-derived chromophore. To sustain vision, vitamin A precursors must be acquired in the gut, metabolized, and distributed to the eyes. Deficiencies in this pathway in inherited ocular disease states deplete cone photoreceptors from chromophore and eventually lead to cell death, whereas the more abundant rod photoreceptors are less affected. However, pathways that support cone function and survival under such conditions are largely unknown. Using biochemical, histological, and physiological approaches, we herein show that intervention with ß-carotene in STRA6-deficient mice improved chromophore supply to cone photoreceptors. Relieving the inherent negative feedback regulation of ß-carotene oxygenase-1 activity in the intestine by genetic means further bolstered cone photoreceptor functioning in the STRA6-deficient eyes. A vitamin A-rich diet, however, did not improve cone photoreceptor function in STRA6-deficiency. We provide evidence that the beneficial effect of ß-carotene on cones results from favorable serum kinetics of retinyl esters in lipoproteins. The respective alterations in lipoprotein metabolism maintained a steady supply of retinoids to the STRA6-deficient eyes, which ameliorated the competition for chromophore between rod and cone photoreceptors. Together, our study elucidates a cone photoreceptor-survival pathway and unravels an unexpected metabolic connection between the gut and the retina.

Reconsidering the residency training pathway for ambulatory care pharmacists.

Ambulatory care, commonly found in primary care settings, is a growing area of pharmacy practice supported by an expanding number of residency training opportunities in this setting. As this practice expands, the profession has chosen to adopt structures that define it as a specialty practice area, a departure from the medical profession. A key example of this definition is the profession's alignment of residency training for this setting within postgraduate year 2 standards. In this commentary, we explore the implications of this approach and share experience from more than 20 years of statewide ambulatory care residency training in Minnesota. We question whether current training expectations are rooted in an objective evaluation of the knowledge and skills required for ambulatory care pharmacy practice. Ultimately, we call on practice leaders to take account of the impact on current training expectations for learners and pharmacy workforce development and seek a rationalization of the training pathway for ambulatory care practice.

ASTER-B regulates mitochondrial carotenoid transport and homeostasis.

The scavenger receptor class B type 1 (SR-B1) facilitates uptake of cholesterol and carotenoids into the plasma membrane (PM) of mammalian cells. Downstream of SR-B1, ASTER-B protein mediates the nonvesicular transport of cholesterol to mitochondria for steroidogenesis. Mitochondria also are the place for the processing of carotenoids into diapocarotenoids by ß-carotene oxygenase-2. However, the role of these lipid transport proteins in carotenoid metabolism has not yet been established. Herein, we showed that the recombinant StART-like lipid-binding domain of ASTER-A and B preferentially binds oxygenated carotenoids such as zeaxanthin. We established a novel carotenoid uptake assay and demonstrated that ASTER-B expressing A549 cells transport zeaxanthin to mitochondria. In contrast, the pure hydrocarbon ß-carotene is not transported to the organelles, consistent with its metabolic processing to vitamin A in the cytosol by ß-carotene oxygenase-1. Depletion of the PM from cholesterol by methyl-ß-cyclodextrin treatment enhanced zeaxanthin but not ß-carotene transport to mitochondria. Loss-of-function assays by siRNA in A549 cells and the absence of zeaxanthin accumulation in mitochondria of ARPE19 cells confirmed the pivotal role of ASTER-B in this process. Together, our study in human cell lines established ASTER-B protein as key player in nonvesicular transport of zeaxanthin to mitochondria and elucidated the molecular basis of compartmentalization of the metabolism of nonprovitamin A and provitamin A carotenoids in mammalian cells.

Exploring the medication-related needs of sports medicine organizations.

BACKGROUND: Medication-related services are provided by sports medicine organizations at all levels of competition; however, no studies have been conducted to date that have been aimed at evaluating the medication-related needs among members of each organization, the challenges associated with meeting these needs, and the potential utilization of pharmacists to facilitate these services for athletes. OBJECTIVES: To explore the medication-related needs within sports medicine organizations and identify where services provided by a pharmacist may assist in reaching organizational goals. METHODS: Qualitative semistructured group interviews were utilized to identify the medication-related needs of sports medicine organizations in the U.S. Organizations, including orthopedic centers, sports medicine clinics, training centers, and athletic departments were recruited via email. A survey and a set of sample questions were sent to each participant to gather demographic information and allow time for the participant to reflect on their specific organization's medication-related needs in preparation for the interviews. A discussion guide was created to explore each organization's overarching medication-related functions and the challenges and successes related to their current policies and procedures regarding their medication-related needs. Each interview was conducted virtually, recorded, and transcribed into text. A thematic analysis was conducted by a primary and secondary coder. Themes and subthemes were identified from the codes and defined. RESULTS: Nine organizations were recruited for participation. Of these, individuals from 3 Division 1 university-based athletic programs were interviewed. Twenty-one individuals participated across all 3 organizations including 16 athletic trainers, 4 physicians, and 1 dietitian. The thematic analysis revealed the following themes: Medication-Related Responsibilities, Barriers to Optimizing Medication Use, Contributions to Successful Implementation of the Medication-Related Services, and Opportunities to Improve Medication-Related Needs. Themes were reduced to subthemes to further describe the medication-related needs within each organization. CONCLUSION: Division 1 university-based athletic programs have medication-related needs and challenges that have the potential to be enhanced by services provided by pharmacists.

Genetic dissection in mice reveals a dynamic crosstalk between the delivery pathways of vitamin A.

Vitamin A is distributed within the body to support chromophore synthesis in the eyes and retinoid signaling in most other tissues. Two pathways exist for the delivery of vitamin A: the extrinsic pathway transports dietary vitamin A in lipoproteins from intestinal enterocytes to tissues, while the intrinsic pathway distributes vitamin A from hepatic stores bound to serum retinol binding protein (RBP). Previously, the intestine-specific homeodomain transcription factor (ISX) and the RBP receptor STRA6 were identified as gatekeepers of these pathways; however, it is not clear how mutations in the corresponding genes affect retinoid homeostasis. Here, we used a genetic dissection approach in mice to examine the contributions of these proteins in select tissues. We observed that ISX deficiency increased utilization of both preformed and provitamin A. We found that increased storage of retinoids in peripheral tissues of ISX-deficient mice was dependent on STRA6 and induced by retinoid signaling. In addition, double-mutant mice exhibited a partial rescue of the Stra6 mutant ocular phenotype. This rescue came at the expense of a massive accumulation of vitamin A in other tissues, demonstrating that vitamin A is randomly distributed when present in excessive amounts. Remarkably, provitamin A supplementation of mutant mice induced the expression of the RBP receptor 2 in the liver and was accompanied by increased hepatic retinyl ester stores. Taken together, these findings indicate dynamic crosstalk between the delivery pathways for this essential nutrient and suggest that hepatic reuptake of vitamin A takes place when excessive amounts circulate in the blood.

LRAT coordinates the negative-feedback regulation of intestinal retinoid biosynthesis from ß-carotene.

There is increasing recognition that dietary lipids can affect the expression of genes encoding their metabolizing enzymes, transporters, and binding proteins. This mechanism plays a pivotal role in controlling tissue homeostasis of these compounds and avoiding diseases. The regulation of retinoid biosynthesis from ß-carotene (BC) is a classic example for such an interaction. The intestine-specific homeodomain transcription factor (ISX) controls the activity of the vitamin A-forming enzyme ß-carotene oxygenase-1 in intestinal enterocytes in response to increasing concentration of the vitamin A metabolite retinoic acid. However, it is unclear how cells control the concentration of the signaling molecule in this negative-feedback loop. We demonstrate in mice that the sequestration of retinyl esters by the enzyme lecithin:retinol acyltransferase (LRAT) is central for this process. Using genetic and pharmacological approaches in mice, we observed that in LRAT deficiency, the transcription factor ISX became hypersensitive to dietary vitamin A and suppressed retinoid biosynthesis. The dysregulation of the pathway resulted in BC accumulation and vitamin A deficiency of extrahepatic tissues. Pharmacological inhibition of retinoid signaling and genetic depletion of the Isx gene restored retinoid biosynthesis in enterocytes. We provide evidence that the catalytic activity of LRAT coordinates the negative-feedback regulation of intestinal retinoid biosynthesis and maintains optimal retinoid levels in the body.

Using a multisite and multi-health system pharmacy resident program model for care documentation quality improvement.

OBJECTIVES: To implement and evaluate a pharmacy resident documentation peer review process. SETTING: The University of Minnesota Postgraduate Year One Pharmacy Residency Program is a multisite program with 25 residents across 16 different health care organizations. PRACTICE DESCRIPTION: Sites within the program provide comprehensive medication management (CMM) services to patients in ambulatory care settings, including participation in the full patient care process of assessment, care plan development, follow-up, and appropriate documentation. PRACTICE INNOVATION: In this innovative peer review process model, residents undergo a deidentified CMM documentation review process with residents from other practice sites, exposing them to different documentation templates and perspectives. EVALUATION: A workgroup of residency preceptors led by a research team developed a peer review process, which evolved through 3 phases over 2 years in response to resident, preceptor, and administration team feedback. Resident feedback was compiled and analyzed. RESULTS: Forty-two residents responded to the survey (67% response rate); 71% found the review process to be helpful. Residents reported that the process improved their understanding of how to improve patient care documentation (74%), how to provide peer feedback (90%), and the importance of effective interprofessional communication in clinical decision making (81%). DISCUSSION: The core perceived benefit of the peer review process was exposure to how other health systems and practitioners document CMM. Some residents participate in a peer review process at their home institutions, which may explain some of the lack of perceived benefit. Generalizability of this study is limited by being within a single residency program with a relatively small number of participants. CONCLUSION: Pharmacy residents found a peer review process of documentation to be helpful during their residency education. The process exposed residents to different documentation practices at various health care systems, which led to ideas of how to improve documentation and provided a foundation for how to provide peer feedback in practice.

Warfarin monitoring in antiphospholipid syndrome and lupus anticoagulant.

OBJECTIVE: To review the available literature on international normalized ratio (INR) and chromogenic factor X (CFX) monitoring in patients with antiphospholipid syndrome (APS), specifically lupus anticoagulant (LA), and furthermore, to identify benefits of one monitoring test compared with the other in the presence of LA. DATA SOURCES: A literature search was conducted through MEDLINE (1946-May 2014) utilizing the following MeSH terms chromogenic compounds, anticoagulants, and factor X. Further articles were identified from original literature citations. STUDY SELECTION: All English-language studies were included that involved INR and/or CFX monitoring in APS patients that focused on a therapeutic anticoagulation level with warfarin therapy. DATA SYNTHESIS: A total of 55 articles were identified, of which nine are referenced because of their relevance for this review: three articles focus on the efficacy of utilizing INR monitoring in patients with APS, five focus on CFX compared with INR for therapeutic warfarin dosing, and one compares different thromboplastins utilizing both INR and CFX monitoring. INR monitoring in patients with APS, specifically LA, was not found to be reliable because thromboplastin reagents are sensitive to LA. Furthermore, when INR was compared to CFX, patients with LA had supratherapeutic INRs despite having CFX within goal range. CONCLUSIONS: In a subgroup of APS patients, INR monitoring may not be safe for determining the dose of warfarin because their INR values can be falsely elevated. Although CFX monitoring is more accurate, it too comes with its own downsides. Managing warfarin therapy in the APS population needs to be individualized.

Promoting diversity, equity and inclusion awareness in clinical documentation through postgraduate year one training.

BACKGROUND AND PURPOSE: As healthcare providers increasingly focus on emerging issues of diversity, equity and inclusion (DEI) in patient care, less is known about the training in postgraduate year one (PGY1) pharmacy residency on DEI clinical documentation considerations. This pilot project explored whether training, discussion and self-reflection within a peer review activity promoted DEI self-awareness in clinical documentation through a centralized curriculum of a multisite PGY1. EDUCATIONAL ACTIVITY AND SETTING: Building upon an established peer review of clinical documentation activity, PGY1 pharmacy residents practicing in ambulatory care settings received training on DEI considerations and completed small and large group discussions, a post-activity mixed methods survey with self-reflection prompts, and a three-month follow-up survey. FINDINGS: Twenty-two residents participated in the peer review of clinical documentation activity, DEI training and discussions. Twelve residents completed the post-activity survey with reflection prompts; 6 (50%) reported similar previous DEI training prior to residency. After the DEI training and discussions, 12 (100%) agreed or strongly agreed that their awareness of DEI documentation considerations increased; 10 (83%) would document their submitted notes differently, while one resident was unsure and one would not make changes. Twelve residents completed the follow-up survey three months following the activity. Themes from the free-text responses on key learnings collected post-activity and three-month post (respectively) included: 1) new knowledge, increased self-awareness, and intended action and 2) increased self-awareness and changes in note-making convention. SUMMARY: Integrating DEI training, discussion, and self-reflection prompts into a peer review clinical documentation activity increased self-awareness and knowledge of DEI considerations and promoted intended changes in patient care documentation for pharmacy residents. Regardless of previous training, residents reported continued self-awareness and changes in documentation conventions continued three months later.

Spirometry: tool for pharmacy practitioners to expand direct patient care services.

OBJECTIVES: To introduce pharmacy practitioners to spirometry testing and provide strategies for integrating this "value-added" tool with other direct patient care pharmacy services. DATA SOURCES: Spirometry literature and resources obtained through search strategies including Ovid, PubMed, and Google Scholar. SUMMARY: Pharmacists are distinctive members of the multidisciplinary patient care team and can contribute by performing spirometry services for pulmonary patients. Pharmacists have been largely absent from performing this much needed service, perhaps as a result of lack of training or because this testing may be perceived as irrelevant to the pharmacist scope of practice. However, pharmacists are actively integrated with many aspects of pulmonary patient care, including recommending and adjusting inhaled pharmacological agents, monitoring for potential drug-drug and drug-disease interactions, recommending smoking cessation, assessing patient prescription insurance coverage, and educating patients, caregivers, and health care providers on use of prescribed respiratory delivery devices. Adding quality spirometry services, based on American Thoracic Society guidelines for accuracy, would increase the breadth and depth of services for pharmacy practitioners. CONCLUSION: Spirometry testing is an added tool for expanding direct patient care pharmacy services. Physician support, appropriate pharmacist training, and understanding of reimbursement of spirometry services are essential in providing quality spirometry testing. Future studies are needed to assess the role of pharmacists in performing spirometry and measuring the performance outcomes of pulmonary patients.

Vitamin A deficiency compromises the barrier function of the retinal pigment epithelium.

A major cause for childhood blindness worldwide is attributed to nutritional vitamin A deficiency. Surprisingly, the molecular basis of the ensuing retinal degeneration has not been well defined. Abundant expression of the retinoid transporter STRA6 in the retinal pigment epithelium (RPE) and homeostatic blood levels of retinol-binding protein delay vitamin A deprivation of the mouse eyes. Hence, genetic dissection of STRA6 makes mice susceptible to nutritional manipulation of ocular retinoid status. We performed RNA-seq analyses and complemented the data with tests of visual physiology, ocular morphology, and retinoid biochemistry to compare eyes with different vitamin A status. Mild ocular vitamin A deficiency decreased transcripts of photoreceptor transduction pathway-related genes and increased transcripts of oxidative stress pathways. The response was associated with impaired visual sensitivity and an accumulation of fluorescent debris in the retina. Severe vitamin A deficiency did not only impair visual perception but also decreased transcripts of genes encoding cell adhesion and cellular junction proteins. This response altered cell morphology, resulted in significant changes in transport pathways of small molecules, and compromised the barrier function of the RPE. Together, our analyses characterize the molecular events underlying nutritional blindness in a novel mouse model and indicate that breakdown of the outer blood-retinal barrier contributes to retinal degeneration and photoreceptor cell death in severe vitamin A deficiency.

Asthma management: How the guidelines compare.

This quick guide details the similarities and differences between recommendations from the National Asthma Education and Prevention Program and the Global Initiative for Asthma.

Addressing practical challenges with entrustable professional activities in ambulatory care experiential education.

BACKGROUND AND PURPOSE: Entrustable professional activities (EPAs) have been adopted and endorsed by the American Association of Colleges of Pharmacy as an assessment strategy. Application of EPAs in ambulatory care advanced pharmacy practice experiences (APPEs) has demonstrated potential internal grading consistency challenges and warrants further guidance. EDUCATIONAL ACTIVITY AND SETTING: Within a required ambulatory care APPE, a rubric was developed using purposeful and convenience sampling of preceptors, faculty, and students to concretely delineate expected EPA level progression from week one to week four based on setting specific activities. FINDINGS: The rubric was perceived to be accurate and useful as a tool for improving expectations of progression in the respective roles on ambulatory care APPEs. SUMMARY: Specifying performance of common clinical activities by weekly EPA level progression within an ambulatory care APPE was perceived to address some of the challenges in APPE grading consistency. Research for determining appropriate progression within an APPE and progression context within graduate and post-graduate training is needed. As more pharmacy training programs implement EPAs, collaboration in applying core EPAs into experiential education will be needed. This example of a setting specific progression rubric may serve as a useful tool for preceptors and students during their APPEs.

Diabetes Aggravates Photoreceptor Pathologies in a Mouse Model for Ocular Vitamin A Deficiency.

Emerging evidence indicates that diabetes disturbs photoreceptor function and vitamin A homeostasis. However, the biochemical basis of this phenotype is not well established. Here, we compared the effects of streptozotocin-induced diabetes in wild-type (WT) mice and Stra6-/- mice, a mouse model for ocular vitamin A deficiency. After 8 weeks, diabetes increased serum retinyl esters in mice of both genotypes. The eyes of diabetic WT mice displayed increased superoxide levels but no changes in retinoid concentrations. Diabetic Stra6-/- mice showed increased ocular retinoid concentrations, but superoxide levels remained unchanged. After 30 weeks, significant alterations in liver and fat retinoid concentrations were observed in diabetic mice. Diabetic WT mice exhibited a decreased expression of visual cycle proteins and a thinning of the photoreceptor layer. Stra6-/- mice displayed significantly lower ocular retinoid concentration than WT mice. An altered retinal morphology and a reduced expression of photoreceptor marker genes paralleled these biochemical changes and were more pronounced in the diabetic animals. Taken together, we observed that diabetes altered vitamin A homeostasis in several organ systems and aggravated photoreceptor pathologies in the vitamin-deficient mouse eyes.

Shifts in Pharmacists' Responsibilities in Family Medicine Residency Programs during COVID-19.

Background: The role of clinical pharmacists in family medicine residency programs (FMRPs) has become increasingly commonplace in the last several years, with mixed responsibilities, however largely involving teaching and patient care. The COVID-19 pandemic affected all facets of healthcare, pharmacists included. Assessment of the impact of COVID-19 on the role of the pharmacist in FMRPs is needed. Methods: A survey tool was developed and distributed through two national listservs clinical pharmacists in FMRPs commonly subscribe to. Results: A total of 32 responses were received. The majority of pharmacist participants indicated no change in their overall time allocation to the FMRP. Patient care was affected by transitions to virtual or remote care for those in the outpatient setting, which in some cases were sustained changes. Teaching responsibilities increased for the majority, largely impacted by the need to transition to virtual platforms. Barriers to relationship development resulted from both of these transitions. Innovations and positive results were reported by participants in patient care, teaching, and research. Conclusions: These results highlight the many silver linings of the COVID-19 pandemic. The new challenges, such as the increased teaching need, use of virtual platforms and using creative ways to find connections and build relationships in an ongoing, semi-virtual world, are opportunities for continued innovation for clinical pharmacists as they fit within our strengths and scope within FMRPs.

Education Standards for Pharmacists Providing Comprehensive Medication Management in Outpatient Nephrology Settings.

Chronic kidney disease is a public health problem that has generated renewed interest due to poor patient outcomes and high cost. The Advancing American Kidney Health initiative aimed to transform kidney care with goals of decreasing the incidence of kidney failure and increasing the number of patients receiving home dialysis or a kidney transplant. New value-based models of kidney care that specify inclusion of pharmacists as part of the kidney care team were developed to help achieve these goals. To support this Advancing American Kidney Health-catalyzed opportunity for pharmacist engagement, the pharmacy workforce must have a fundamental knowledge of the core principles needed to provide comprehensive medication management to address chronic kidney disease and the common comorbid conditions and secondary complications. The Advancing Kidney Health through Optimal Medication Management initiative was created by nephrology pharmacists with the vision that every person with kidney disease receives optimal medication management through team-based care that includes a pharmacist to ensure medications are safe, effective, and convenient. Here, we propose education standards for pharmacists providing care for individuals with kidney disease in the outpatient setting to complement proposed practice standards.

Molecular components affecting ocular carotenoid and retinoid homeostasis.

The photochemistry of vision employs opsins and geometric isomerization of their covalently bound retinylidine chromophores. In different animal classes, these light receptors associate with distinct G proteins that either hyperpolarize or depolarize photoreceptor membranes. Vertebrates also use the acidic form of chromophore, retinoic acid, as the ligand of nuclear hormone receptors that orchestrate eye development. To establish and sustain these processes, animals must acquire carotenoids from the diet, transport them, and metabolize them to chromophore and retinoic acid. The understanding of carotenoid metabolism, however, lagged behind our knowledge about the biology of their receptor molecules. In the past decades, much progress has been made in identifying the genes encoding proteins that mediate the transport and enzymatic transformations of carotenoids and their retinoid metabolites. Comparative analysis in different animal classes revealed how evolutionary tinkering with a limited number of genes evolved different biochemical strategies to supply photoreceptors with chromophore. Mutations in these genes impair carotenoid metabolism and induce various ocular pathologies. This review summarizes this advancement and introduces the involved proteins, including the homeostatic regulation of their activities.

HIV-1 Tat and Morphine Differentially Disrupt Pyramidal Cell Structure and Function and Spatial Learning in Hippocampal Area CA1: Continuous versus Interrupted Morphine Exposure.

About half the people infected with human immunodeficiency virus (HIV) have neurocognitive deficits that often include memory impairment and hippocampal deficits, which can be exacerbated by opioid abuse. To explore the effects of opioids and HIV on hippocampal CA1 pyramidal neuron structure and function, we induced HIV-1 transactivator of transcription (Tat) expression in transgenic mice for 14 d and co-administered time-release morphine or vehicle subcutaneous implants during the final 5 d (days 9-14) to establish steady-state morphine levels. Morphine was withheld from some ex vivo slices during recordings to begin to assess the initial pharmacokinetic consequences of opioid withdrawal. Tat expression reduced hippocampal CA1 pyramidal neuronal excitability at lower stimulating currents. Pyramidal cell firing rates were unaffected by continuous morphine exposure. Behaviorally, exposure to Tat or high dosages of morphine impaired spatial memory Exposure to Tat and steady-state levels of morphine appeared to have largely independent effects on pyramidal neuron structure and function, a response that is distinct from other vulnerable brain regions such as the striatum. By contrast, acutely withholding morphine (from morphine-tolerant ex vivo slices) revealed unique and selective neuroadaptive shifts in CA1 pyramidal neuronal excitability and dendritic plasticity, including some interactions with Tat. Collectively, the results show that opioid-HIV interactions in hippocampal area CA1 are more nuanced than previously assumed, and appear to vary depending on the outcome assessed and on the pharmacokinetics of morphine exposure.

Carotenoid metabolism at the intestinal barrier.

Carotenoids exert a rich variety of physiological functions in mammals and are beneficial for human health. These lipids are acquired from the diet and metabolized to apocarotenoids, including retinoids (vitamin A and its metabolites). The small intestine is a major site for their absorption and bioconversion. From here, carotenoids and their metabolites are distributed within the body in triacylglycerol-rich lipoproteins to support retinoid signaling in peripheral tissues and photoreceptor function in the eyes. In recent years, much progress has been made in identifying carotenoid metabolizing enzymes, transporters, and binding proteins. A diet-responsive regulatory network controls the activity of these components and adapts carotenoid absorption and bioconversion to the bodily requirements of these lipids. Genetic variability in the genes encoding these components alters carotenoid homeostasis and is associated with pathologies. We here summarize the advanced state of knowledge about intestinal carotenoid metabolism and its impact on carotenoid and retinoid homeostasis of other organ systems, including the eyes, liver, and immune system. The implication of the findings for science-based intake recommendations for these essential dietary lipids is discussed. This article is part of a Special Issue entitled Carotenoids recent advances in cell and molecular biology edited by Johannes von Lintig and Loredana Quadro.