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Kinetoplastid organisms, including Trypanosoma brucei, are a significant health burden in many tropical and semitropical countries. Much of their metabolism is poorly understood. To better study kinetoplastid metabolism, chemical probes that inhibit kinetoplastid enzymes are needed. To discover chemical probes, we have developed a high-throughput flow cytometry screening assay that simultaneously measures multiple glycolysis-relevant metabolites in live T. brucei bloodstream form parasites. We transfected parasites with biosensors that measure glucose, ATP, or glycosomal pH. The glucose and ATP sensors were FRET biosensors, while the pH sensor was a GFP-based biosensor. The pH sensor exhibited a different fluorescent profile from the FRET sensors, allowing us to simultaneously measure pH and either glucose or ATP. Cell viability was measured in tandem with the biosensors using thiazole red. We pooled sensor cell lines, loaded them onto plates containing a compound library, and then analyzed them by flow cytometry. The library was analyzed twice, once with the pooled pH and glucose sensor cell lines and once with the pH and ATP sensor cell lines. Multiplexing sensors provided some internal validation of active compounds and gave potential clues for each compound's target(s). We demonstrated this using the glycolytic inhibitor 2-deoxyglucose and the alternative oxidase inhibitor salicylhydroxamic acid. Individual biosensor-based assays exhibited a Z'-factor value acceptable for high-throughput screening, including when multiplexed. We tested assay performance in a pilot screen of 14,976 compounds from the Life Chemicals Compound Library. We obtained hit rates from 0.2 to 0.4% depending on the biosensor, with many compounds impacting multiple sensors. We rescreened 44 hits, and 28 (64%) showed repeatable activity for one or more sensors. One compound exhibited EC 50 values in the low micromolar range against two sensors. We expect this method will enable the discovery of glycolytic chemical probes to improve metabolic studies in kinetoplastid parasites., Competing Interests: Declaration of competing interest All authors declare that they have no conflicts of interest., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Hypermobile Ehlers-Danlos syndrome (hEDS) is a common heritable connective tissue disorder that lacks a known genetic etiology. To identify genetic contributions to hEDS, whole exome sequencing was performed on families and a cohort of sporadic hEDS patients. A missense variant in Kallikrein-15 (KLK15 p. Gly226Asp), segregated with disease in two families and genetic burden analyses of 197 sporadic hEDS patients revealed enrichment of variants within the Kallikrein gene family. To validate pathogenicity, the variant identified in familial studies was used to generate knock-in mice. Consistent with our clinical cohort, Klk15 G224D/+ mice displayed structural and functional connective tissue defects within multiple organ systems. These findings support Kallikrein gene variants in the pathogenesis of hEDS and represent an important step towards earlier diagnosis and better clinical outcomes.

The rising costs of healthcare, increased chronic illnesses, and healthcare provider burnout has led to an environment desperate for scalable solutions to ease practice burdens. With a projected shortage in the number of primary healthcare providers available to provide team-based care, community-based pharmacy practitioners are accessible and eager to assist. In order to provide enhanced patient care services to aid their clinician colleagues, community-based pharmacists will have to transform their practices to support the provision of enhanced services and medication optimization in value-based payment models. The purpose of this article is to define how multiple factors in pharmacy, healthcare, technology and payment models aligned to create an opportunity for the Community Pharmacy Foundation and CPESN® USA to implement a nationwide community pharmacy practice model called 'Flip the Pharmacy'. This new model aims to scale community pharmacy practice transformation and move beyond filling prescriptions at a moment-in-time to caring for patients over time through a 24-month step-wise program paired with in-person pharmacist coaching. Preliminary observations from the first six months of the program highlight community pharmacy as a site of care with community-based pharmacist practitioners providing and documenting targeted patient care interventions., (© Individual authors.)

Study Objective: To assess the effectiveness of a community pharmacist-delivered cardiovascular case-management program by comparing body mass index (weight), systolic and diastolic blood pressure, and full lipid profile at the beginning of the program with these outcome measures at the end of the program., Design: Retrospective data analysis using billing data submitted between July 1, 2001, and October 31, 2004, with a pre-post design in which subjects served as their own controls., Setting: Manufacturing workplace in rural Iowa., Participants: Fifty-six workers with risk factors for cardiovascular disease (mean age 40.67 yrs), 37 had diabetes mellitus and 19 did not., Intervention: During visits to the workers, pharmacists provided education about cardiovascular disease, identification of drug therapy problems, and importance of routine blood pressure, pulse, and weight measurements; they communicated with participants' physicians as needed., Measurements and Main Results: The number of pharmacist visits/participant ranged from 1-13 (mean +/- SD 6.97 +/- 3.05). Outcome measures were weight, systolic and diastolic blood pressures, full lipid profiles (in patients with diabetes), and percentage of patients achieving treatment goal by the end of the 3 years. Statistically significant differences between the first and last visits were achieved for both systolic (124.12 +/- 11.07 and 120.36 +/- 14.39 mm Hg, respectively, p=0.016) and diastolic (80.4 +/- 9.01 and 77.43 +/- 9.14 mm Hg, respectively, p=0.019) blood pressure. The 19 patients without diabetes showed a statistically significant improvement in diastolic blood pressure (p=0.039), but the 37 patients with diabetes did not show a significant difference. A nonsignificant increase was seen in the percentage of patients with diabetes achieving low-density lipoprotein cholesterol (LDL) level goal between the first and last visits (p=0.06)., Conclusion: A cardiovascular case-management program delivered in the workplace to middle-aged working adults by community pharmacists improved blood pressure and reduced LDL levels. The program was not effective, however, in weight reduction.

Study Objective: To determine whether community pharmacists can use point-of-service health status assessments to identify and resolve drug-related problems (DRPs) in ambulatory patients with selected musculoskeletal (MSK) disorders., Design: Twelve-month, prospective, multicenter demonstration project., Setting: Twelve independent community pharmacies in eastern Iowa., Patients: Ambulatory patients with self-reported diagnosis of osteoarthritis, rheumatoid arthritis, or low back pain., Measurements: During quarterly pharmacy visits for 1 year, patients used touch-screen computers to report their health status. Patients answered questions on the Short Form-36 (SF-36) general health survey, as well as questions assessing limitations associated with their MSK condition. Pharmacists used this data in interviewing patients to assess for DRPs., Main Results: The study enrolled 461 patients, of whom 388 returned for the 12-month visit. During this 1-year period, community pharmacists identified 926 cumulative DRPs. Patients with no DRPs had significantly higher physical component summary scores on the SF-36 (p<0.05) than patients with more than one DRP at baseline (36.2 vs 31.6), 6 months (39.2 vs 33.3), and 12 months (40.1 vs 35.4). At 12 months, actions performed by pharmacists led to resolution or improvement of 70.7% of DRPs., Conclusion: Drug-related problems are numerous in community-dwelling patients with MSK disorders and correspond to decreased physical health status. Community pharmacists can use patient-reported measures of health status to identify DRPs and initiate processes to resolve them.

In the past decade, the pharmacy profession has made remarkable strides in implementing a wide range of pharmacy-based patient care services. To foster greater awareness of the value of these services among payers and to ensure the long-term success of pharmaceutical care, pharmacists need to focus more attention on obtaining compensation for these services. In the long run, pharmacists are likely to receive greater net profits from pharmaceutical care than from dispensing. As Norwood et al. noted, pharmacies keep all the revenues they receive from pharmaceutical care as profits and to cover operating expenses, whereas they keep only about 29% of the revenues from the sale of products. By exploring innovative markets for pharmaceutical care services and continuing to improve rates of reimbursement from third party payers, pharmacists can further enhance the revenues they obtain from their growing array of patient care services.