Now showing items 41-60 of 1438

    • A patient-centered approach to evaluating the role of medical cannabis in the treatment of chronic pain

      Basappa, Sapna; Liang, Connie; Tucker, Shannon; Pincus, Kathleen J. (2022-12)
    • Advancing Regulatory Science to Assure COAs Reflect Diverse Patient Experiences

      Mullins, C. Daniel; Hall, Candace; Kakim, Saya; Medeiros, Michelle; Perfetto, Eleanor M. (2023-10-22)
    • The role of a cysteine residue within an ERK1/2 substrate docking site on signaling and proliferation of melanoma cells containing BRAF mutations

      Grogan, Lena; McClean, Nathaniel; Shapiro, Paul, Ph.D. (2023-10-13)
      The role of extracellular signal-regulated kinase 1/2 (ERK1/2) in signaling pathways in cells is crucial for cell proliferation. Within specific types of cancers, a member of this pathway, BRAF, is mutated at the valine (V)600 position so that this pathway is continuously activated, leading to uncontrolled proliferation. A docking site in ERK1/2 is of interest for inhibitors to control activation of downstream proteins responsible for transcription.1 A compound has been developed to target a substrate docking site and was found to target a specific cysteine2. This residue has been mutated via CRISPR CAS9 in both ERK1 and ERK2 and the proposed studies investigate the effects the ERK1/2 cysteine mutations have on A375 cell melanoma cells regarding cell signaling and proliferation.
    • Engagement of Older People and Caregivers as Partners in Medication Optimization Patient-Centered Outcomes Research

      Tang, Viviane; Haynes, Jodi-Ann; Wang, Sabrina; Lee, Merton; Isetts, Brian; Brandt, Nicole J.; Cooke, Catherine E.; Elonge, Eposi (2023-10-27)
    • Zinc Fingers are a General Target for Persulfidation by Hydrogen Sulfide (H2S)

      Stoltzfus, Andrew T.; Vigane, Thibaut; Li, Haoju; Worth, Madison M.; Filipovic, Milos R.; Michel, Sarah L.J. (2023-10-13)
      Persulfidation (or "sulfhydration") of protein thiols by the gasotransmitter hydrogen sulfide (H2S) has been recently established as an important signaling event associated with oxidative stress and cellular aging. While H2S, mostly bisulfide (HS-) anion at physiological pH, likely does not persulfidate protein thiols directly due to incompatible electric potential, it may act through small- molecule thiols (ex: glutathione) to add a sulfur to cysteine thiols (CysSH → CysSSH). This highly reactive persulfide species can alter protein function and/or scavenge intracellular radical species during oxidative stress by direct reaction with radicals. Zinc finger proteins (ZFs) are potential targets for this PTM as they contain Cys-rich zinc-binding domains and our laboratory has previously reported the direct reaction of the ZF protein TTP with H2S. This reaction requires O2 and involves in-situ persulfidation. To understand how general ZF persulfidation is, we applied a persulfide- specific proteomics approach and observed a trend between Cys content of ZF domains and frequency of persulfidation. A series of TTP variants were prepared and analyzed for H2S reactivity via cryo-electrospray ionization mass spectrometry, as well as UV-visible, circular dichroism, and fluorescence spectroscopies. We found that all peptide variants bound Zn(II) and were persulfidated by H2S to some extent, with higher Cys content contributing to greater persulfide labeling and ROS in the -CCCH and -CCCC peptides. Current work is focused on proteomic classification of persulfidated ZFs and elucidating the radical mechanism of this PTM using chemical tags, fluorescence, and spin-traps.
    • Capsule 2023

      University of Maryland, Baltimore. School of Pharmacy, 2023
    • Investigating the Role of an ERK1/2 Cysteine Mutation on MAP Kinase Signaling Pathways

      Piluk, Arianna; Jones, Kristine; Gudivada, Himaja; McClean, Nathaniel; Grogan, Lena; Jateng, Danielle; Shapiro, Paul, Ph.D. (2023-07-28)
    • Development of an In-Cell Footprinting Method Coupled with MS for the Study of Proteins in Three-Dimensional Cancer Models

      Shortt, Raquel; Wang, Hongbing; Jones, Lisa M. (2023)
      Fast photochemical oxidation of proteins (FPOP) is a powerful, mass spectrometry (MS)-based, biophysical method used to probe protein structure, interactions, and conformations. FPOP was recently extended into cells (IC-FPOP) and can modify thousands of proteins in a single experiment, enabling proteome-wide structural biology. Although IC-FPOP can reveal critical structural information in 2D cell culture, the conditions do not emulate an in-vivo environment. To address this, we propose to develop a mass spectrometry-based protein footprinting method that assesses the varying protein heterogeneity in 3D cell culture; Spheroid-FPOP. IC-FPOP on intact spheroids was performed using a patented PIXY platform which brought automation to IC-FPOP. Spheroid-FPOP coupled with serial trypsinization to obtain spatial resolution, revealed modifications in three distinct spheroid regions; the outer inner and core. Native oncogenic pathways were interrogated through this study showing its value in disease pathogenesis and treatment. Though progressive for FPOP, the extension into 3D model systems generated three times the samples and data compared to typical IC- or IV-FPOP experiments. This shed light to FPOP workflow limitations. The research herein responds to those challenges by developing an automated sample preparation workflow by coupling a sample handling robot with Thermo’s sample preparation kit. These modifications robustly improve the workflow by significantly reducing the manual labor, execution time, and variability of samples processed and data acquired. After workflow optimization FPOP, we apply the optimized method more complex biological sample. In our case, 3D bioprinted Huh-7 liver organoids were generated for IC-FPOP. To obtain spatial resolution within the model, we integrated cryosectioning of the top, middle and bottom layers of the organoid. Peptide level analysis revealed differences in the extent of modification for peptides identified in each region of the organoid, which confirms the acquisition of structural information. However further optimization was required to increase proteome depth. By coupling the organoid model with IC-FPOP we aim further validate its implementation for complex proteome-wide structural studies. In all, this research is focused on advancing the applications and processing workflows for IC-FPOP.
    • The Effect of Medication Information Delivery Format on Cognitive Load and Knowledge Retention of Informal Caregivers

      McPherson, Mary Lynn M.; Kulo, Violet A; Cestone, Christina (2023)
      Informal caregivers (IFCs) are tasked with many responsibilities in patient care, including medication management. Many IFCs feel ill-prepared for this responsibility, and it is incumbent on health care professionals to provide education and ensure IFCs competence in medication management. One common strategy is to provide a medication information leaflet to the IFC to prepare them for this role. Designing medication information leaflets using sound educational principles, such as an infographic designed according to the cognitive theory of multimedia learning (CTML), may optimize knowledge retention and decrease cognitive load for IFCs. The purpose of this randomized, experimental study was to investigate the impact of medication information delivery format on immediate retention of medication information and cognitive load of IFCs of patients with a serious illness. Using purposive sampling, 120 IFCs who have provided some element of medication management for patients diagnosed with a serious illness, including patients who may have been receiving hospice or palliative care services were recruited. Study participants were randomly assigned in either the experimental group or the control group. The experimental group viewed an infographic on the medication hydromorphone, followed by a knowledge quiz, and a self-assessment of cognitive load. This was followed by a second infographic on hydroxyzine, the quiz, and cognitive load assessment. The control group went through the same steps but viewed a text-only medication leaflet. Statistical analyses included descriptive statistics, independent samples t-test, one-way analysis of variance, and one-way multivariate analysis of variance. Statistically significantly higher quiz scores were observed among those who viewed the infographics than those who viewed text-only medication leaflets, indicating better immediate knowledge retention of medication information. Those who viewed the infographic also had statistically significantly lower intrinsic and extraneous cognitive load, and higher germane cognitive load. These findings are consistent with the hypothesis that infographics prepared using the CTML result in better and more efficient learning. Limitations of this research include use of nonprobability sampling, examining only two medications that are commonly used in serious illness, and lack of systematic randomization. Additional research is needed to continue determining best practices for instructing and supporting IFCs in medication management. 
    • The Effects of Graded Versus Ungraded Individual Readiness Assurance Tests on Pharmacy Students’ Assessment Performance and Achievement Goals in a Team-Based Learning Classroom

      Noel, Zachary; Cestone, Christina; Gordes, Karen L. (2023)
      Individual readiness assurance tests (iRATs) are frequently graded in team-based learning (TBL) classrooms, with the goal of incentivizing individual pre-class preparation. The purpose of this study was to determine whether shifting to an ungraded iRAT process affects student preparation and learning, as measured using assessment scores, and whether this is accompanied by a change in achievement goals. Using a crossover design in a required second-year Doctor of Pharmacy pharmacotherapy course, students were assigned to one of two iRAT grading sequences: graded/ungraded (G/UG) or ungraded/graded (UG/G). In the G condition iRATs were graded based on correctness and in the UG condition based on completion. Each period consisted of four iRATs and one examination. Students completed the Achievement Goal Questionnaire at the conclusion of each period. A one-way repeated measures multivariate analysis of variance (MANOVA) was used to test within-subject differences of mean iRAT and examination scores across grading conditions. A separate one-way repeated measures MANOVA was used to analyze differences in achievement goal scores. A total of 91 doctor of pharmacy students were included in the study. There was a statistically significant main effect for iRAT grading condition on assessment scores, F(2,88) = 3.851, Wilks’ Λ = .992, p = .025. Univariate testing using one-way analysis of variance with Bonferroni correction demonstrated a significant difference only in iRAT scores, with the mean score higher in the G condition (72.51% versus 67.99%; p = .011). Examination scores were similar in the G and UG conditions (81.07% versus 80.32%, p = .397). There was not a statistically significant difference in achievement goals based on iRAT grading condition, F(4,85) = 1.109, η2 =.050, p = .358. In conclusion, a modest reduction in iRAT performance was observed when shifting from a graded to ungraded iRAT; however, this had no effect on examination performance. Achievement goals were unaffected by the change in iRAT grading condition.
    • KRSC-University of Maryland's CERSI MOU Signing Ceremony

      Maryland's Center of Excellence in Regulatory Science and Innovation; Korean Regulatory Science Center; Korean Ministry of Food and Drug Safety (2023-06-05)