Inhibition of FAAH, TRPV1, and COX2 by NSAID-serotonin conjugates.

Serotonin was linked by amidation to the carboxylic acid groups of a series of structurally diverse NSAIDs. The resulting NSAID-serotonin conjugates were tested in vitro for their ability to inhibit FAAH, TRPV1, and COX2. Ibuprofen-5-HT and Flurbiprofen-5-HT inhibited all three targets with approximately the same potency as N-arachidonoyl serotonin (AA-5-HT), while Fenoprofen-5-HT and Naproxen-5-HT showed activity as dual inhibitors of TRPV1 and COX2.

Evaluating the effects of a mindfulness mobile application on student pharmacists' stress, burnout, and mindfulness.

PURPOSE: Pharmacists report high levels of burnout. Mindfulness approaches have been demonstrated to have positive results in the general population and in other healthcare professions. However, limited studies have been performed evaluating mindfulness approaches in student pharmacists. The aim of this study was to evaluate the effectiveness of daily use of a mindfulness mobile application in improving student pharmacists' perceived stress, burnout, and mindfulness. METHODS: This study was a randomized, longitudinal, waitlist-controlled trial. The intervention group was asked to meditate using the mindfulness application Headspace daily for at least 6 weeks. The waitlist control group was asked to abstain from using the application for the entire study. Stress, burnout, and mindfulness were assessed using validated survey instruments at baseline, 6 weeks, and 10 weeks. A secondary outcome was to assess the persistence of application use after the intervention period. RESULTS: Fifty-six participants completed the study. The intervention group reported significantly lower scores on stress and burnout at 6 weeks compared to the control group. The intervention group also reported significantly higher scores on mindfulness. The differences in stress, burnout, and mindfulness persisted at follow-up. The mean percentage of students in the intervention group who used the application each day was 90% over the intervention period and 62% over the follow-up period. CONCLUSION: A mindfulness mobile application significantly improved student pharmacists' stress, burnout, and mindfulness with daily use. Most participants continued to use the application for 4 weeks after the end of the intervention. Positive effects on stress and mindfulness persisted even with decreased use.

Getting Rigor Right.

A colleague of mine once shared a story he heard of an organic chemistry teacher who would tell people he had one of the most important, lifesaving jobs in the world. Why? Because he made sure academically unprepared premed students never made it to medical school. Teachers can have many possible motivations for maintaining high academic standards in their courses. A desire to ensure future health practitioners are adequately prepared to engage in the intellectual rigor of their profession may be just one of them.

Top consumer uses of bestselling, single-ingredient vitamin and mineral supplements.

OBJECTIVE: Vitamin and mineral supplements are widely used for self-care of a variety of medical conditions, but little is known about the specific conditions for which they are used. This study mined consumer product reviews to determine specific ways vitamin and mineral supplements are being used therapeutically. DESIGN/SETTING: A cross-sectional analysis of user reviews for top-selling, single-ingredient vitamin and mineral products from a popular online retailer was performed to identify the most frequently appearing words associated with medical conditions. Results of individual analyses were compared to achieve consensus on the top, relevant keywords for each supplement. The full text of the reviews was searched for these keywords to distinguish whether they referred to therapeutic uses or adverse effects. RESULTS: A total of 14 vitamin and 11 mineral supplements were analyzed. The number of user reviews for the analyzed products varied from 41 for manganese to over 5000 for biotin and vitamin D (median = 547 reviews per product). Cohen's kappa test for investigator-selected keywords related to medical conditions was generally greater than 0.6, indicating good interrater reliability. From these lists, the top consumer self-care uses were identified for 24 supplements. Commonly reported adverse effects were also noted for several products. CONCLUSION: This study used data mining to identify the top ways consumers use an array of bestselling, single-ingredient vitamin and mineral supplements. These results can provide healthcare and nutrition professionals with information to anticipate the supplement-related education needs of patients and provide researchers with priority areas for clinical studies.

A Pilot Evaluation of I'M HAPPY: An Interactive Module to Halt Abuse of Prescriptions in Preteens and Youth.

Objective: The purpose of this project was to create a web-accessible drug education program that would positively impact student knowledge and perceptions. Materials and Methods: An interactive computer module (Interactive Module to Halt Abuse of Prescriptions in Preteens and Youth [I'M HAPPY]) was created using a combination of e-learning and animation software. The module used narrated illustrations, animations, and games to explain what over-the-counter (OTC) and prescription drugs are, how they are used correctly, and what risks they carry. Animated drug refusal scenarios with user-determined outcomes were also incorporated. Sixth-grade students were asked to complete the module online and were given a 14-question pretest and posttest on the definitions, adverse effects, and proper use of OTC and prescription medicines. They were also asked to rank the perceived helpfulness of several drug refusal strategies. Student opinions about the computer module itself were collected on the posttest. Results: Students (n = 34) scored better on quiz questions following the module. They also showed increased favorability toward three of four drug refusal strategies after completing the module, with a greater proportion ranking these strategies "Very Helpful." Most students indicated they learned new things from the module (79%) and found it enjoyable (88%) and easy to use (97%). Conclusion: The I'M HAPPY program improved student knowledge of OTC and prescription drugs and enhanced perceptions of the helpfulness of certain drug refusal strategies. The computer module was considered enjoyable and easy to use, and can be easily disseminated to any student or classroom having internet access.

Lysophosphatidic acid is constitutively produced by human peritoneal mesothelial cells and enhances adhesion, migration, and invasion of ovarian cancer cells.

Lysophosphatidic acid (LPA) is both a potential marker and a therapeutic target for ovarian cancer. It is critical to identify the sources of elevated LPA levels in ascites and blood of patients with ovarian cancer. We show here that human peritoneal mesothelial cells constitutively produce LPA, which accounts for a significant portion of the chemotactic activity of the conditioned medium from peritoneal mesothelial cells to ovarian cancer cells. Both production of LPA by peritoneal mesothelial cells and the chemotactic activity in the conditioned medium can be blocked by HELSS [an inhibitor of the calcium-independent phospholipase A(2) (iPLA(2))] and AACOCF(3) [an inhibitor of both cytosolic PLA(2) (cPLA(2)) and iPLA(2)]. Moreover, cell-based enzymatic activity assays for PLA(2) indicate that peritoneal mesothelial cells have strong constitutive PLA(2) activity. Receptors for LPA, LPA(2), and LPA(3) are involved in the conditioned medium-induced chemotactic activity. Invasion of ovarian cancer cells into peritoneal mesothelial cells has also been analyzed and shown to require PLA(2), LPA receptors, and the mitogen-activated protein/extracellular signal-regulated kinase kinase/extracellular signal-regulated kinase signaling pathway. Thus, we show here, for the first time, that human peritoneal mesothelial cells constitutively produce bioactive lipid signaling molecules, such as LPA, via iPLA(2) and/or cPLA(2) activities. Conditioned medium from peritoneal mesothelial cells stimulate migration, adhesion, and invasion of ovarian cancer cells, and may play similar roles in vivo.

Lessons Learned Using a Demonstration in a Large Classroom of Pharmacy Students.

Objective. To create a demonstration that would enhance student understanding of the following acid/base concepts: the opposing effects of a weak acid and its conjugate base, the acidifying effects of carbon dioxide on blood pH, and the resistance of a buffered solution to changes in pH. Methods. The demonstrations were delivered to four different years of first-year pharmacy (P1) students in three different ways: as live in-class demonstrations, as a series of three short videos posted online, and as a series of three short videos used both in class and posted online. Quizzes were administered to each set of students and qualitative feedback solicited. Comparisons were made between the four classes that received the demonstrations and with two previous classes that had not received the demonstrations but had been given the same quiz questions. Results. Student feedback indicated difficulty seeing and understanding the live demonstrations, and the demonstration videos were not widely used when they were posted online. However, students provided positive feedback on demonstration videos when they were used in class in addition to being posted online. Using videos, performance increased on quiz items related to the learning objectives of the demonstrations, returning to levels seen in years prior to the decline in student performance. Conclusion. When delivered appropriately, a series of pH demonstrations appeared to improve student assessment performance.

Using drawings to probe the affective experience of pharmacy students on rotation: A pilot study.

INTRODUCTION: The objective of this study was to use drawings to assess the global affective experience of pharmacy students on rotation. METHODS: Students nearing graduation were asked to submit a drawing that captured the essence of their Advanced Pharmacy Practice Experience (APPE). Drawings were analyzed qualitatively using inductive content analysis and the affects portrayed in the drawings were coded, themed and categorized. RESULTS: Of the 22 (21% response rate) submissions, four categories emerged as positive, negative, bisemous (can be positive or negative), and mixed. DISCUSSION: The results from the study demonstrated a range of feelings such as easiness, inadequateness, busyness, tiredness, learning, and excitement. The drawings also appeared superficially to fit well into the flow model of subjective experience. CONCLUSION: Drawings by pharmacy students provided insight into their feelings about their rotations. Further studies are needed to determine how this method can be utilized to record affective experiences and how they can be used to improve experiential learning.

Synthesis and evaluation of fluorogenic substrates for phospholipase D and phospholipase C.

Fluorogenic analogues of phosphatidylcholine and lysophosphatidylcholine, DDPB and lysoDDPB, were synthesized by an enzyme-assisted strategy. The analogues were evaluated as substrates for phospholipases C and D and lysophospholipase D. DDPB was cleaved by bacterial and plant phospholipase D (PLD) enzymes and represents the first direct fluorogenic substrate for real-time measurement of PLD activity. Both fluorogenic substrates have potential in screening for PLD and PC-PLC inhibitors and for monitoring spatiotemporal changes in PLD activity in cells. [structure: see text]

A board game to assist pharmacy students in learning metabolic pathways.

OBJECTIVES: To develop and evaluate a board game designed to increase students' enjoyment of learning metabolic pathways; their familiarity with pathway reactions, intermediates, and regulation; and, their understanding of how pathways relate to one another and to selected biological conditions. DESIGN: The board game, entitled Race to Glucose, was created as a team activity for first-year pharmacy students in the biochemistry curriculum. ASSESSMENT: A majority of respondents agreed that the game was helpful for learning regulation, intermediates, and interpathway relationships but not for learning reactions, formation of energetic molecules, or relationships, to biological conditions. There was a significant increase in students' scores on game-related examination questions (68.8% pretest vs. 81.3% posttest), but the improvement was no greater than that for examination questions not related to the game (12.5% vs. 10.9%). CONCLUSION: First-year pharmacy students considered Race to Glucose to be an enjoyable and helpful tool for learning intermediates, regulation, and interpathway relationships.

Fluorogenic phospholipids as head group-selective reporters of phospholipase A activity.

PLA (phospholipases A) are important mediators of cell signaling, generating bioactive fatty acids and LPLs (lysophospholipids). PLA products having different head groups can initiate vastly different types of signaling. Fluorogenic analogues of the PLs (phospholipids) PA (phosphatidic acid), PC (phosphatidylcholine), PE (phosphatidylethanolamine), and PG (phosphatidylglycerol) were synthesized as PLA substrates for rapidly determining in real time the influence of head group modifications on cell signaling both in vitro and in cells. Enzyme-assisted remodeling of the sn-2 position of the diacylglyceryl moiety with cobra venom PLA 2 and transphosphatidylation with a particular PLD (phospholipase D) were central steps in the preparation of these enzymatic probes. The resulting fluorogenic Dabcyl- and BODIPY-containing PL analogues, DBPA, DBPC, DBPE, and DBPG, were used in mixed micelle assays to determine PLA 2 kinetics. Next, the assays were used to determine the X i (50) value of a common PLA 2 inhibitor. Finally, the head group selectivities of a series of commercially available PLA 2 enzymes were readily established using the DBPLs (Dabcyl-BODIPY PLs) as substrates.

Caspase-3-dependent activation of calcium-independent phospholipase A2 enhances cell migration in non-apoptotic ovarian cancer cells.

Calcium-independent phospholipase A(2) (iPLA(2)) plays a pivotal role in phospholipid remodeling and many other biological processes, including inflammation and cancer development. iPLA(2) can be activated by caspase-3 via a proteolytic process in apoptotic cells. In this study we identify novel signaling and functional loops of iPLA(2) activation leading to migration of non-apoptotic human ovarian cancer cells. The extracellular matrix protein, laminin-10/11, but not collagen I, induces integrin- and caspase-3-dependent cleavage and activation of overexpressed and endogenous iPLA(2). The truncated iPLA(2) (amino acids 514-806) generates lysophosphatidic acid and arachidonic acid. Arachidonic acid is important for enhancing cell migration toward laminin-10/11. Lysophosphatidic acid activates Akt that in turn acts in a feedback loop to block the cleavage of poly-(ADP-ribose) polymerase and DNA fragmentation factor as well as prevent apoptosis. By using pharmacological inhibitors, blocking antibodies, and genetic approaches (such as point mutations, dominant negative forms of genes, and siRNAs against specific targets), we show that beta(1), but not beta(4), integrin is involved in iPLA(2) activation and cell migration to laminin-10/11. The role of caspase-3 in iPLA(2) activation and cell migration are supported by several lines of evidence. 1) Point mutation of Asp(513) (a cleavage site of caspase-3 in iPLA(2)) to Ala blocks laminin-10/11-induced cleavage and activation of overexpressed iPLA(2), whereas mutation of Asp(733) to Ala has no such effect, 2) treatment of inhibitors or a small interfering RNA against caspase-3 results in decreased cell migration toward laminin-10/11, and 3) selective caspase-3 inhibitor blocks cleavage of endogenous iPLA(2) induced by laminin-10/11. Importantly, small interfering RNA-mediated down-regulation of endogenous iPLA(2) expression in ovarian carcinoma HEY cells results in decreased migration toward laminin, suggesting that our findings are pathophysiologically important.