Specific in situ immuno-imaging of pulmonary-resident memory lymphocytes in human lungs.

Introduction: Pulmonary-resident memory T cells (TRM) and B cells (BRM) orchestrate protective immunity to reinfection with respiratory pathogens. Developing methods for the in situ detection of these populations would benefit both research and clinical settings. Methods: To address this need, we developed a novel in situ immunolabelling approach combined with clinic-ready fibre-based optical endomicroscopy (OEM) to detect canonical markers of lymphocyte tissue residency in situ in human lungs undergoing ex vivo lung ventilation (EVLV). Results: Initially, cells from human lung digests (confirmed to contain TRM/BRM populations using flow cytometry) were stained with CD69 and CD103/CD20 fluorescent antibodies and imaged in vitro using KronoScan, demonstrating it's ability to detect antibody labelled cells. We next instilled these pre-labelled cells into human lungs undergoing EVLV and confirmed they could still be visualised using both fluorescence intensity and lifetime imaging against background lung architecture. Finally, we instilled fluorescent CD69 and CD103/CD20 antibodies directly into the lung and were able to detect TRM/BRM following in situ labelling within seconds of direct intra-alveolar delivery of microdoses of fluorescently labelled antibodies. Discussion: In situ, no wash, immunolabelling with intra-alveolar OEM imaging is a novel methodology with the potential to expand the experimental utility of EVLV and pre-clinical models.

Comparison of Student Pharmacists' Knowledge Retention Utilizing Electronic Health Records Versus Simultaneously Completing Simulated Experiences.

Objective. The purpose of this study was to determine whether the addition of standardized patients or the addition of interprofessional student teams with standardized patients to the use of a simulated electronic health record improved student knowledge retention and perceptions.Methods. This was a prospective cohort study assessing three cohorts of first-year student pharmacists in pharmacy skills laboratory activities that occurred in 2018, 2019, and 2021. The primary objective of the study was to compare knowledge retention of the case material between groups at one month. Each year, an element of simulated experience was added onto the previous year's case. In 2018, students completed the case using only the electronic health record web application. In 2019, the previous year's experience was combined with an objective structured clinical examination (OSCE) with standardized patients. In 2021, the 2019 experience was supplemented with student physician assistants. Case scores and student perceptions were also compared between groups.Results. Of the 260 potential participants, 238 students were included in the primary analysis. Results showed that with the addition of interprofessional team-based care and standardized patients, significant improvement was demonstrated in knowledge retention assessments at one month. Mean knowledge retention assessment scores for the 2018, 2019, and 2021 groups were 63.8%, 71.7%, and 76.1%, respectively. Significant improvement was also found in student perceptions.Conclusion. Adding standardized patients and interprofessional team-based care to a pharmacy skills laboratory that uses a simulated electronic health record significantly improved student knowledge retention and perceptions.

Current cytochrome P450 phenotyping methods applied to metabolic drug-drug interaction prediction in dogs.

Recombinant cytochrome P450 (P450) phenotyping, different approaches for estimating fraction metabolized (f(m)), and multiple measures of in vivo inhibitor exposure were tested for their ability to predict drug interaction magnitude in dogs. In previous reports, midazolam-ketoconazole interaction studies in dogs have been attributed to inhibition of CYP3A pathways. However, in vitro phenotyping studies demonstrated higher apparent intrinsic clearances (CL(int,app)) of midazolam with canine CYP2B11 and CYP2C21. Application of activity correction factors and isoform hepatic abundance to liver microsome CL(int,app) values further implicated CYP2B11 (f(m) >or= 0.89) as the dog enzyme responsible for midazolam- and temazepam-ketoconazole interactions in vivo. Mean area under the curve (AUC) in the presence of the inhibitor/AUC ratios from intravenous and oral midazolam interaction studies were predicted well with unbound K(i) and estimates of unbound hepatic inlet inhibitor concentrations and intestinal metabolism using the AUC-competitive inhibitor relationship. No interactions were observed in vivo with bufuralol, although significant interactions with bufuralol were predicted with fluoxetine via CYP2D and CYP2C pathways (>2.45-fold) but not with clomipramine (<2-fold). The minor caffeine-fluvoxamine interaction (1.78-fold) was slightly higher than predicted values based on determination of a moderate f(m) value for CYP1A1, although CYP1A2 may also be involved in caffeine metabolism. The findings suggest promise for in vitro approaches to drug interaction assessment in dogs, but they also highlight the need to identify improved substrate and inhibitor probes for canine P450s.

Feasibility of implementing a reduced fasting protocol for critically ill trauma patients undergoing operative and nonoperative procedures.

BACKGROUND: This prospective, observational cohort study was designed to determine the feasibility of implementing a reduced enteral fasting protocol in mechanically ventilated trauma patients undergoing selected operative and nonoperative procedures. METHODS: Critically ill, mechanically ventilated trauma patients undergoing selected operative and nonoperative procedures received enteral nutrition up until the time of the procedure, if receiving small bowel feeds, or received enteral nutrition that was discontinued 45 minutes before the procedure, if receiving gastric feeds. RESULTS: Measures of delivery of nutrition such as total enteral nutrition delivered and days required to reach nutrition goal were collected. Complications measured were death, incidence of ventilator-associated pneumonia, urinary tract infection, catheter-related bloodstream infection, wound infection, hypoglycemia, and emesis during procedures. No significant demographic differences were observed between the 2 groups. Patients in the intervention group showed trends toward greater total enteral nutrition delivered and faster attainment of target nutrition goals, although these measures were not statistically significant. Patients in the intervention group had rates of infective complications similar to those in the standard group. The median (interquartile range) for intensive care unit length of stay in the intervention group vs standard group was 7 (5, 15) vs 7 (5, 12) (P = 0.94), and the ventilator days were 8 (4.2, 14) vs 7 (3, 11) (P = 0.37). CONCLUSIONS: A reduced fasting protocol was feasible for selected operative procedures, with trends toward improving nutrition delivery and no increase in adverse outcomes. A larger randomized study of this approach is warranted before adoption of this practice can be advocated.

Impact of Standardized Simulated Patients on First-Year Pharmacy Students' Knowledge Retention of Insulin Injection Technique and Counseling Skills.

Objective. To compare pre- and post-intervention test scores assessing insulin injection technique and counseling skills among P1 students with (intervention) or without (control) simulated patients, and to compare counseling checklist and knowledge retention test scores between groups. Methods. This study utilized cluster randomization. In addition to traditional instruction, the intervention group counseled a simulated patient on the use of insulin using the teach-back method. Test score changes from baseline were analyzed via two-sample t-test. Results. The intervention group exhibited a significantly greater increase in knowledge test scores from baseline compared to the control group. Similar changes were seen in post-instruction counseling checklist scores and knowledge retention test scores from baseline. Conclusion. Simulated patient interactions, when added to traditional coursework within a P1 skills lab, improve student counseling aptitude and knowledge retention scores.

The quality of life of hospice patients: patient and provider perceptions.

The purposes of this study were to describe the quality of life (QOL) of terminally ill patients in a home-based hospice program and to examine the relationship between QOL data and patients' symptom distress, ability to function, interpersonal communication (support from family and friends), well-being (their affairs in order), and transcendence (religious comfort/support) as recorded in their charts. QOL was measured by the Missoula-Vitas Quality of Life Index (MVQOLI), an instrument designed specifically for use with terminally ill patients. The study was conducted over a three-year period with 129 terminally ill patients enrolled in a home-based hospice program of care. The MVQOLI was administered to patients within 20 days of their admission to hospice. A retrospective chart review was conducted to determine patients' levels of symptom distress, ability to function, social support, whether or not their affairs were in order, and religious comfort/support. The mean age of participants in this study was 67, with 54.3 percent male and 45.7 percent female. Cancer was the primary diagnosis for 92.2 percent of the sample, and 35 percent of these patients had a diagnosis of lung cancer. Of the 7.8 percent non-cancer diagnoses, five were diagnosed with AIDS, four with chronic obstructive pulmonary disease, and one with chronic heart failure. The results of this study revealed positive scores on the five dimensions of the MVQOLI QOL scale, indicating that within 20 days of admission to hospice, patients rated their QOL as good to very good. Data obtained from the chart review also indicated that patients did not experience a great deal of symptom distress (e.g., pain, nausea, shortness of breath, and restlessness). A significant correlation existed between age and QOL; number of interventions and pain levels; and marital status, well-being, interpersonal relationships, and transcendence. Shortness of breath and well-being were significantly correlated with QOL. There was no significant correlation between gender, race, or closeness to death and the five dimensions of the MVQOLI and chart review assessments.

Patient and caregiver satisfaction with end-of-life care: does high satisfaction mean high quality of care?

The purpose of this study was to determine patient and caregiver satisfaction with a hospice program of care. The setting for the study was a home-care hospice in the southeastern United States that provides a full range of services for patients with life-limiting illness and supportive services for family caregivers. Two Likert-type instruments were used to determine satisfaction with staff, communication, education, information provided, symptom management, promptness with service, and overall satisfaction. Some 321 patients and 443 caregivers completed surveys over a two-year period of time. Data indicates the majority of patients and their caregivers were very satisfied with hospice services and the care they received. Providing quality care at the end of life is the goal of hospice. Satisfaction with delivery of care, management of symptoms, and communication with staff are all components of quality care and contribute to quality of life.