Emotional and Metacognitive Impacts of a Human Anatomy Course Utilizing Three-Dimensional Learning Modalities

A high school human anatomy course was designed to integrate three-dimensional learning modalities through the use of three-dimensional visualization technology, augmented reality, and whole-body donors (cadaveric specimens), and customized 3D digital models were created to support student learning. The course was organized so that each class meeting fell into one of six unique learning session types: lectures, augmented reality labs, gross anatomy (cadaveric) labs, content application (active learning) sessions, review sessions, and examination sessions. An interval scale using ten emoji, each associated with a particular emotional valence (i.e., pleasantness) level, was developed and integrated into the course to track students’ emotional states before and after each learning session. Results indicated that students generally felt neutral or pleasant emotions and experienced significant valence level shifts over a variety of learning session types. Additionally, some correlations between their emotional valence levels and their exam scores were identified, and students experienced statistically significant changes in their emotions regarding learning in a cadaveric laboratory over the span of the course. Four summative lecture and laboratory unit examinations were developed to assess the students’ abilities to remember and to understand information derived from unit learning objectives, using four unique question types. Student metacognitive shifts were analyzed by evaluating the alignment of a students’ pre-examination confidence level, measured on a four-factor scale, for a particular learning objective to a students’ post-response certainty level, measured on a four-factor scale, for a question assessing that particular learning objective. Knowledge levels were defined and could fall into one of four general categories: complete, partial, absent, or flawed, although more nuanced sub-categories were defined and analyzed. Results indicated that students performed best, were most confident, and were most certain on questions utilizing screenshots of 3D digital models, whereas students performed worst, were least confident, and were least certain on questions utilizing cadaveric specimens. Additionally, there were more complete knowledge levels and fewer absent knowledge levels acquired on the unit lecture examinations, as compared to the unit laboratory examinations

Translational Pathways for Ultra Long-Acting Medicines for Chronic Disease: From HIV to Substance Use Disorders

Human immunodeficiency virus (HIV) and opioid use disorder (OUD) are among the top global public health challenges, with a significant overlap in the number of cases and higher mortality risks compared to the general population. Treatment and recreational opioid use lead to addictive disorders. Both conditions require repeated dosing of therapy where adherence to a strict lifelong regimen remains a major challenge. Notably, people living with HIV/AIDS (PLWHA) and struggling with opioid use disorders (OUD) show suboptimal adherence to antiretroviral drugs (ARVs). Also, the risk of transmitting HIV and other blood-borne infections through the sharing of needles and other injection equipment is high in patients with OUD. Recent studies have demonstrated that initiation of long-acting treatments among HIV and OUD patients is not only associated with decreased risk for opioid-related adverse events but also improved viral suppression. We hypothesized that the creation of ultra-long-acting (ULA) prodrug formulations could further improve treatment outcomes. We, therefore, sought to develop ultra-long-acting formulations of a broadly used integrase strand transfer inhibitor called dolutegravir (DTG) and mu-opioid receptor (MOR) partial agonist called buprenorphine (BUP), which has demonstrated a significant advantage in reducing opioid overdose deaths. For DTG, a library of monomeric and dimeric prodrug nanoformulations was synthesized and subjected to in vitro and preclinical pharmacokinetic (PK) screening studies that led to the identification of the lead 18-carbon chain-modified ester prodrug nanocrystal candidate (coined NM2DTG) with the potential to sustain therapeutic drug concentrations for over six months after a single intramuscular (IM) dose. Ideal physiochemical and PK properties facilitated slow DTG release from tissue macrophage depot stores at the muscle injection site and adjacent lymphoid tissues following single parenteral injections. Significant plasma drug levels were recorded for up to a year in rodents following a single injection of NM2DTG. Tissue sites for prodrug hydrolysis were dependent on nanocrystal dissolution and prodrug release, drug-depot volume, perfusion, and cell-tissue pH. Each affected an extended NM2DTG apparent half-life recorded by PK parameters. Similarly, a single IM injection of the lead BUP nanoformulations in Sprague Dawley rats sustains plasma BUP levels at or above the target mu-opioid receptor occupancy concentrations for over three months in ongoing PK studies. We hypothesize that BUP prodrugs will prolong the drug’s apparent half-life and allow for creation of organic solvent free ULA formulations. In summary, the development of ultra-long-acting DTG and BUP injectable formulations can improve treatment of HIV and OUD by enhancing adherence and delivery of the two broadly used drugs into sites of action

Genetic Drivers and Tumor Milieu Analysis in Mantle Cell Lymphoma

Mantle Cell Lymphoma (MCL) is a B-cell lymphoma characterized by t(11;14) leading to CyclinD1 (CCND1) overexpression with highly variable clinical response. Although the tumor intrinsic mechanisms have been fairly studied, the role of the tumor microenvironment (TME) in survival and therapeutic responses is marginally examined. Herein, we integrated high-throughput genomics and characterized TME using a highly multiplexed, spatially resolved, single-cell digital pathology approach termed imaging mass cytometry (IMC) to delineate the tumor-immune landscape. We identified eight MCL genomic subtypes with distinct signatures and prognoses. Subsets C2, C5, and C6 had poorer outcomes with a 5-year overall survival (OS) of 33-50% and a median OS of 3.2-4.5 years. In contrast, subtypes C1, C3, C4, C7, and C8 showed better survival, with a 5-year OS of 62-88% and a median OS of 8.2-11.8 years. Subtypes with complex genomic alterations, like deletions and mutations involving TP53 and regions on chromosomes 13, 17, and 11, correlated with shorter survival. Conversely, subtypes with gains in MYC, mutation/deletion of ATM, mutations in SP140, and deletions on chromosome 6 associated with longer survival, broadly categorizing the subtypes into two groups based on survival outcomes. Extrinsic factors such as TME reveal nine major cellular compartments, with neoplastic cells constituting 58% and various immune cell types forming the tumor milieu. Notably, TP53-altered genetic subtypes were characterized by an increase in T-regulatory, CD8+ T-cells, and endothelial cells, alongside heightened exhaustion markers on immune and neoplastic cells. Poor prognostic indicators included elevated PDL-2 or low HLADR on CD8+ T cells and high CD45RA on SOX11+/- malignant cells. Spatial analysis identified distinct cellular neighborhoods correlated with TP53 alterations. Thus, to further explore the underlying pathobiology of this most lethal TP53 subtype in MCL, we generated modified cell lines to study the functional significance of p53 deficiency in MCL. We v performed RNA-seq on these p53-modified cells to determine their transcriptomic features. Notably, we found that BCR signaling was remarkably repressed upon WT p53 expression, suggesting that p53 deficiency may contribute to resistance against BTK inhibitor treatment. Using the CUT&RUN assay to determine the TP53 targets, besides canonical targets, we observed changes in an expansive list of genes and pathways upon p53 modification, including the BCR signaling. Interestingly, PTPN6, targeted by TP53 and a negative regulator of BCR, modulated BCR pathway activity which may potentially alter ibrutinib responsiveness. Thus, this study suggests a more granular risk stratification recovering previously known subtypes and identifying novel subgroups. TME analysis of the TP53 altered subgroup demonstrates that, far from being histopathologically monotonous, MCL has a complex tumor-immune architecture, and that changes in tumor topology can be correlated with clinically relevant features. This analysis identified candidate biomarkers and therapeutic targets such as TIM-3, PD-L2, HLA-DR that are relevant for combination treatment strategies in immuno-oncology and cellular therapies. Loss of PTPN6 could act as an alternative biomarker in patients with p53 functional loss likely to respond to ibrutinib treatment indicating pharmacological inhibition of PTPN6 might be a novel approach to enhance the efficacy of BCR-targeted therapies

Metabolic Diversity of Human Macrophages: Potential Influence on Staphylococcus Aureus Intracellular Survival

Staphylococcus aureus is a leading cause of medical device-associated biofilm infections. This is influenced by the ability of S. aureus biofilm to evade the host immune response, which is partially driven by the anti-inflammatory cytokine interleukin-10 (IL-10). Here, we show that treatment of human monocyte-derived macrophages (HMDMs) with IL-10 enhanced biofilm formation, suggesting that macrophage anti-inflammatory programming likely plays an important role during the transition from planktonic to biofilm growth. To identify S. aureus genes that were important for intracellular survival in HMDMs and how this was affected by IL-10, transposon sequencing was performed. The size of the S. aureus essential genome was similar between unstimulated HMDMs and the outgrowth control (18.5% vs 18.4%, respectively, with 54.4% overlap) but increased to 22.5% in IL-10-treated macrophages, suggesting that macrophage polarization status exerts differential pressure on S. aureus. Essential genes for S. aureus survival within IL-10-polarized HMDMs were dominated by negative regulatory pathways, including nitrogen and RNA metabolism, whereas S. aureus essential genes within untreated HMDMs were enriched in biosynthetic pathways such as purine and pyrimidine biosynthesis. To explore how IL-10 altered the macrophage intracellular metabolome, targeted metabolomics was performed on HMDMs from six individual donors. IL-10 treatment led to conserved alterations in distinct metabolites that were increased (dihydroxyacetone phosphate, glyceraldehyde-3-phosphate, and acetyl-CoA) or reduced (fructose-6-phosphate, aspartic acid, and ornithine) across donors, whereas other metabolites were variable. Collectively, these findings highlight an important aspect of population-level heterogeneity in human macrophage responsiveness that should be considered when translating results to a patient population.IMPORTANCEOne mechanism that Staphylococcus aureus biofilm elicits in the host to facilitate infection persistence is the production of the anti-inflammatory cytokine interleukin-10 (IL-10). Here, we show that exposure of human monocyte-derived macrophages (HMDMs) to IL-10 promotes S. aureus biofilm formation and programs intracellular bacteria to favor catabolic pathways. Examination of intracellular metabolites in HMDMs revealed heterogeneity between donors that may explain the observed variability in essential genes for S. aureus survival based on nutrient availability for bacteria within the intracellular compartment. Collectively, these studies provide novel insights into how IL-10 polarization affects S. aureus intracellular survival in HMDMs and the importance of considering macrophage heterogeneity between human donors as a variable when examining effector mechanisms

Non-TNFi Biologic and Targeted Synthetic DMARDs in Rheumatoid Arthritis-Associated Interstitial Lung Disease: A Comparative, Propensity Score-Matched Study Using National Veterans Affairs Data

Background/Objectives: There is a paucity of evidence on the safety and effectiveness of non- TNFi biologic therapies in RA-ILD, this study aimed to compare treatment outcomes in RA-ILD between rituximab, abatacept, IL-6i, and JAKi using the Target Trial Emulation Framework. Methods: We emulated 3 trials comparing abatacept, IL-6i, and JAKi with rituximab (reference). Patients fulfilling validated RA-ILD algorithms initiating one of these non-TNFi b/tsDMARDs were propensity score (PS)-matched (1:1) using national Veterans Affairs (VA) data from 2006 to 2020. PS models included demographics, comorbidities, general health status indicators, and RA- and ILD-related severity measures. Study outcomes were death and respiratory-related hospitalization, ascertained by VA data and linkages to the National Death Index and Medicare, over 3-year and 1-year follow-up periods. Cox regression models with univariate frailty correction were used to analyze study outcomes. Sensitivity analyses were performed among cohorts with modified eligibility criteria. Results: In the primary cohort, we 1:1 matched abatacept (n=159), IL-6i (n=78), and JAKi (n=94) with rituximab initiators [age (mean range 68.1-69.9 years), male (range 87-91%), white (range 78-84%)]. There was not a significant difference in the primary composite outcome among any of the comparisons [abatacept = HR: 0.90 [0.64, 1.26]; IL-6i = HR: 0.97 [0.62, 1.53], aHR: 0.93 [0.59, 1.47]; JAKi= HR: 0.85 [0.53,1.36], aHR: 0.69 [0.41, 1.14]]. Sensitivity analyses supported the findings. Conclusions: We did not find significant differences in outcomes between non-TNFi b/tsDMARDs, though estimates were imprecise. These findings emphasize the need for clinical trials of advanced immunomodulatory therapies in RA-ILD

Evaluation of a Student Experiential Learning Clinic for Hand Therapy Using a Logic Model

Student clinics (SC) provide experiential learning opportunities in occupational therapy (OT) education that develop clinical reasoning, while providing much needed rehabilitation to under and un-insured patients in the community. The Student Experiential Learning Clinic for Hand Therapy (SELC-HT) is a SC that used a logic model for planning, implementing, and evaluating the SELC-HT. The purpose of this study is to report on outcome data on students and patients, as outlined in the evaluation phase of the logic model. The 13 OT master/doctorate students, who delivered care in the SELC-HT, demonstrated growth in self-reported hand therapy knowledge (p=0.002) measured with the Hand Therapy Certification Commission Self-Assessment Tool. Nine of the 12 students responding to alumni survey were employed in hand therapy positions shortly after graduation. Five students authored six manuscripts published in peer-reviewed journals or practice journals about their work in the SELC-HT. Of the 57 patients with baseline data, fractures were the most common diagnosis, and most patients were Black and males. One-third of injuries were due to violence, primarily gunshot wounds. At discharge (n=25) mean disability, measured with the Disability Arm Shoulder Hand, decreased 14.8 points (p=.001), which exceeds minimal clinical difference of 10.83. Statistically significant improvements in work disability (n=18) and work ability (n=21) also occurred. Most importantly, five patients who were not able to work at baseline had returned to work at discharge. These positive student and patient outcomes are due in part to the systematic planning and implementation of procedures defined in the SELC-HT logic model

Online & Blended Learning Environments: A Mixed Methods Study Evaluating the Feasibility, Acceptability, & Influence of an Online Professional Development Course for Health Professions Educators

Online and blended teaching and learning (OBTL) are integral to the future and success of higher education, including health professions education. Institutions with a goal of developing high-quality online and blended programs must prioritize time and resources dedicated to professional development and training. This research study utilized an online professional development course as an intervention to gain deeper insights into the scope of faculty development to effectively teach in online and blended learning environments. The study, utilizing a convergent mixed methods approach, gathered data through a pre- and post- intervention survey measuring health professions faculty readiness to teach online, a knowledge-based test, and post-intervention focus group discussions. Statistically significant differences in survey and test scores were observed between pre- and post-intervention and advantages of the intervention were highlighted in the focus group discussions. These key findings suggest the efficacy and influence of the educational intervention. Additionally, barriers and recommendations for enhancement were identified, including a notable gap between perceived importance and perceived capability among faculty members. This information pinpoints areas where professional development and support may be beneficial. Outcomes of the study provide valuable insights into health professions faculty members\u27 knowledge, readiness, and perceptions related to teaching in online or blended educational environments

The Effect of Repetitive Transcranial Magnetic Stimulation on Brain and Cognitive Variables in Patients with Amnestic Mild Cognitive Impairment

Healthy and pathological aging processes can impact human memory. Current interventions for clinical memory impairment offer little opportunity for restoration of memory ability. Fortunately, a novel therapy in the form of repetitive transcranial magnetic stimulation (rTMS) has exhibited promise in this regard. In healthy adults, rTMS of the angular gyrus (AG) has been reported to improve memory. The current study sought to determine the generalizability of these findings to individuals with clinical memory impairment by administering a clinical trial replicating an rTMS protocol reported to improve hippocampal-dependent memory in healthy individuals to individuals with amnestic mild cognitive impairment (aMCI). Individuals diagnosed with aMCI (N=15) completed two five-day sessions of rTMS with a minimum two-week washout between weeks. Brain and cognitive variables were collected before the first rTMS session of each week and the day following the final session of each week. rTMS was administered as a β-frequency pulse sequence alternating 2-sec. trains of 20 Hz stimulation with 28-sec. rest for 20 min (1600 TMS pulses total) for five consecutive days. Stimulation was administered to each participant\u27s area of left AG exhibiting peak resting-state functional connectivity (RSFC) with left hippocampus. Treatment rTMS was delivered at 100% resting motor threshold (RMT) and placebo rTMS at 10% RMT. This study followed a double-blind crossover design. The primary objective of this study was to identify any changes in hippocampal-dependent memory associated with rTMS. Secondary objectives included identifying RSFC changes of the hippocampus and AD with the DMN and any changes in the brain\u27s functional connectome associated with rTMS. Treatment rTMS was associated with significantly improved performance on select measures of hippocampal-dependent memory, specifically verbal memory. Treatment rTMS also reduced RSFC between the hippocampus and bilateral dorsolateral prefrontal cortex. However, no change in connectomic measures was observed within this study. This study\u27s findings suggest that rTMS can improve memory and modify hippocampal RSFC in individuals diagnosed with aMCI and may serve as a potential intervention for AD

Using Evidenced-Based Practices to Improve Sonography Scan Lab Instruction: Peyton’s 4-Step Approach

Objective: The purpose of this study was to examine the effects of implementing Peyton’s 4-Step Approach for skills teaching of sonography laboratory instruction in the first semester of a diagnostic medical sonography (DMS) program. Materials & Methods: Participants included DMS students enrolled at UNMC during the fall semester of academic years 2020 and 2021. Data was collected through pre- and post-lab surveys completed by the DMS students. During weekly scan lab sessions, DMS faculty modeled Peyton’s 4-Step Approach to skills training: Step 1: Demonstration, Step 2: Deconstruction, Step 3: Comprehension, and Step 4: Performance. Results: The pre-lab survey found that no DMS students (n=0/26) had prior experience with Peyton’s 4-Step Approach for skills teaching. The post-lab survey found 88% (n=15/17) of DMS students agreed Peyton’s 4-Step Approach to lab instruction aligned with their learning style. For Step 1 (demonstration), 82% (n=14/17) of DMS students routinely watched the pre-lab video prior to lab and 82% (n=14/17) found the videos helpful in learning the scan lab skills. 88% (n=15/17) agreed the live, instructor demonstration with spoken description in Step 2 (deconstruction) was helpful. For Step 3 (comprehension), 88% (n=15/17) agreed the live instructor demonstration guided by student spoken description was helpful. Of the steps used in Peyton\u27s approach, 88% (n=15/17) responded that Step 4 (performance) was most beneficial, while 12% (n=2/17) responded that Step 3 (comprehension) was most beneficial. Conclusion: Implementation of Peyton’s Approach improved sonography lab instruction in the first semester of a DMS program. This technique standardized teaching processes and DMS students found all steps to be helpful in learning lab skills. While performance or “hands on” step was considered the most helpful, students found all steps were beneficial for learning sonography skills