An Online, Self-directed Pharmacy Bridging Course for Incoming First-Year Students.

Objective. To evaluate the short-term effectiveness of an online bridging course to increase the knowledge of struggling incoming students' in crucial content areas within the Doctor of Pharmacy (PharmD) curriculum. Methods. An assessment was administered to all incoming first-year pharmacy students (N=180) during orientation to determine their foundational knowledge in key areas. Students who scored <70% on the assessment (N=137) were instructed to complete a 10-module, online, self-directed bridging course focusing on physiology, biochemistry, math, and medical terminology during the first two weeks of the quarter to prepare them for first-quarter coursework. After completing the bridging course, participants completed the same assessment to determine content knowledge acquisition and retention. At the end of the quarter, the assessment was again administered to all first-year students, regardless of whether they had completed the bridging course. Results. The average assessment score of students who completed the bridging course modules improved significantly (53% vs 76%). All students demonstrated significant improvement in assessment scores between orientation and the end of the quarter; however, bridging course participants achieved a greater increase in assessment scores (53% vs 73%) than nonparticipants (76% vs 81%). Significant relationships were found between assessment scores following completion of the bridging course and pass rates in first-quarter courses. Conclusion. The online, self-directed bridging course offered at Midwestern University, Chicago College of Pharmacy proved successful as a method of knowledge acquisition and as a system for early identification (within the first two weeks of the quarter) of students in need of additional academic support.

Inclusion of calcium during isolation of high-density lipoprotein from plasma maintains antioxidant function.

We investigated how inclusion of calcium during isolation of high-density lipoprotein (HDL) affected its antioxidant function. Following isolation, HDL was dialyzed against 0.154 M NaCl without or with added calcium (1mM). HDL's paraoxonase 1 activity was unaffected by calcium treatment (87 ± 11% of normal vs. 89 ± 16% of normal, P=0.826). In contrast, whereas HDL dialyzed with calcium inhibited oxidation of low-density lipoprotein (LDL) by 87 ± 10%, HDL dialyzed without calcium inhibited oxidation by only 58 ± 19% (P=0.004). Thus, inclusion of calcium during isolation is important for maintaining HDL's antioxidant function.

Critical thinking and reflection exercises in a biochemistry course to improve prospective health professions students' attitudes toward physician-pharmacist collaboration.

OBJECTIVE: To determine the impact of performing critical-thinking and reflection assignments within interdisciplinary learning teams in a biochemistry course on pharmacy students' and prospective health professions students' collaboration scores. DESIGN: Pharmacy students and prospective medical, dental, and other health professions students enrolled in a sequence of 2 required biochemistry courses. They were randomly assigned to interdisciplinary learning teams in which they were required to complete case assignments, thinking and reflection exercises, and a team service-learning project. ASSESSMENT: Students were asked to complete the Scale of Attitudes Toward Physician-Pharmacist Collaboration prior to the first course, following the first course, and following the second course. The physician-pharmacist collaboration scores of prospective health professions students increased significantly (p<0.001). CONCLUSIONS: Having prospective health professions students work in teams with pharmacy students to think and reflect in and outside the classroom improves their attitudes toward physician-pharmacist collaboration.

Interprofessional workshop to improve mutual understanding between pharmacy and medical students.

OBJECTIVE: To measure changes in pharmacy and medical students' physician-pharmacist collaboration scores resulting from a workshop designed to promote understanding of the others' roles in health care. METHODS: More than 88% of first-year pharmacy (n = 215) and medical (n = 205) students completed the Scale of Attitudes Toward Physician-Pharmacist Collaboration on 3 occasions in order to establish a baseline of median scores and to determine whether the scores were influenced by an interprofessional workshop. RESULTS: Participation in the interprofessional workshop increased pharmacy students' collaboration scores above baseline (p=0.02) and raised the scores of medical students on the education component of the collaboration survey instrument (p=0.015). The collaboration scores of pharmacy students greatly exceeded those of medical students (p<0.0001). CONCLUSION: A workshop designed to foster interprofessional understanding between pharmacy and medical students raised the physician-pharmacist collaboration scores of both. Crucial practical goals for the future include raising the collaboration scores of medical students to those of pharmacy students.

Vitamin C attenuates hypochlorite-mediated loss of paraoxonase-1 activity from human plasma.

Paraoxonase 1 (PON1) is a cardioprotective enzyme associated with high-density lipoprotein (HDL). We tested the hypothesis that vitamin C protects HDL and PON1 from deleterious effects of hypochlorous acid, a proinflammatory oxidant. In our experiments, HDL (from human plasma) or diluted human plasma was incubated with hypochlorite in either the absence (control) or presence of vitamin C before measuring chemical modification and PON1 activities. Vitamin C minimized chemical modification of HDL, as assessed by lysine modification and accumulation of chloramines. In the absence of vitamin C, chloramines accumulated to 114 +/- 4 micromol/L in HDL incubated with a 200-fold molar excess of hypochlorite; but addition of vitamin C (200 micromol/L) limited formation to 36 +/- 6 micromol/L (P < .001). In plasma exposed to hypochlorite, IC(50) values of 1.2 +/- 0.1, 9.5 +/- 1.0, and 5.0 +/- 0.6 mmol/L were determined for PON1's phosphotriesterase, arylesterase, and (physiologic) lactonase activities, respectively. Vitamin C lessened this inhibitory effect of hypochlorite on PON1 activities. In plasma supplemented with vitamin C (400 micromol/L), PON1 phosphotriesterase activity was 72% +/- 17% of normal after incubation with hypochlorite (2 mmol/L), compared with 42% +/- 6% for unsupplemented plasma (P < .05). Similar effects were seen for other PON1 activities. In some experiments, vitamin C also appeared to reverse hypochlorite-mediated loss of PON1 phosphotriesterase activity; but this effect was not observed for the other PON1 activities. In conclusion, vitamin C attenuated hypochlorite-mediated loss of PON1 activity in vitro and may, therefore, preserve cardioprotective properties of HDL during inflammation.

Modification with homocysteine does not increase susceptibility of human low-density lipoprotein to iron-mediated oxidation.

Oxidation of human low-density lipoprotein (LDL) is centrally involved in the development of cardiovascular diseases. This study investigated whether homocysteine-mediated thiolation of LDL rendered it more susceptible to oxidation by iron. After in vitro exposure to homocysteine thiolactone for 60 minutes, LDL's thiol content increased from 26 +/- 5 (control) to 224 +/- 20 nmol/mg of protein (thiolated; P < .0001). Control and thiolated LDL (0.2 mg of protein per milliliter) were incubated with either redox active iron (Fe(3+); 10 micromol/L) or, as a positive control, copper (Cu(2+); 10 micromol/L). Consistent with the observation of others, thiolation decreased Cu(2+)-dependent formation of lipid oxidation products in LDL (17 +/- 16 nmol/mg of protein formed in thiolated LDL, compared with 81 +/- 21 nmol/mg of protein in control, during 6 hours of incubation; P < .01). Thiolation had no effect, however, on Fe(3+)-mediated oxidation of LDL with lipid oxidation products remaining essentially nondetectable during prolonged incubation (up to 48 hours). Thiolation similarly had no effect on oxidation of LDL (0.2 mg of protein per milliliter) by heme-complexed iron (hemin; 10 micromol/L), with lipid oxidation products increasing to 24 +/- 1 and 27 +/- 4 nmol/mg of protein for control and thiolated LDL, respectively, during 6 hours of incubation (P > .05). Similar results were observed using LDL with varying degrees of thiolation (29 +/- 5, 85 +/- 14, 130 +/- 15, and 213 +/- 19 nmol of thiol per milligram of protein). In conclusion, these results demonstrate that thiolation has no effect on LDL's susceptibility to iron-mediated oxidation.

Vitamin C preserves the cardio-protective paraoxonase activity of high-density lipoprotein during oxidant stress.

HDL-associated paraoxonase (PON) antioxidant enzyme activity is cardio-protective. We investigated whether vitamin C prevented loss of PON activity from HDL during oxidant stress. HDL was incubated with either hydrophilic or lipophilic peroxyl radical initiators in the absence (control) or presence of vitamin C (50 and 100 micromol/L). Regardless of the type of radical, accumulation of lipid oxidation products in HDL was similar in incubations lacking vitamin C. Loss of PON activity was greater in HDL exposed to hydrophilic, in contrast to lipophilic, radicals, but addition of vitamin C maintained enzyme activity. Vitamin C's capacity to attenuate loss of the HDL ability to prevent atherogenic modification of LDL (assessed as electrophoretic mobility) was, however, modest, and appeared limited only to those incubations in which HDL was exposed to lipophilic radicals. Our results indicate that vitamin C may, under some conditions, prevent loss of cardio-protective function from HDL during oxidant stress.

Differential modulation of microglia superoxide anion and thromboxane B2 generation by the marine manzamines.

BACKGROUND: Thromboxane B2 (TXB2) and superoxide anion (O2-) are neuroinflammatory mediators that appear to be involved in the pathogenesis of several neurodegenerative diseases. Because activated-microglia are the main source of TXB2 and O2- in these disorders, modulation of their synthesis has been hypothesized as a potential therapeutic approach for neuroinflammatory disorders. Marine natural products have become a source of novel agents that modulate eicosanoids and O2- generation from activated murine and human leukocytes. With the exception of manzamine C, all other manzamines tested are characterized by a complex pentacyclic diamine linked to C-1 of the beta-carboline moiety. These marine-derived alkaloids have been reported to possess a diverse range of bioactivities including anticancer, immunostimulatory, insecticidal, antibacterial, antimalarial and antituberculosis activities. The purpose of this investigation was to conduct a structure-activity relationship study with manzamines (MZ) A, B, C, D, E and F on agonist-stimulated release of TXB2 and O2- from E. coli LPS-activated rat neonatal microglia in vitro. RESULTS: The manzamines differentially attenuated PMA (phorbol 12-myristate 13-acetate)-stimulated TXB2 generation in the following order of decreasing potency: MZA (IC50 < 0.016 microM) > MZD (IC50 = 0.23 microM) > MZB (IC50 = 1.6 microM) > MZC (IC50 = 2.98 microM) > MZE and F (IC50 > 10 microM). In contrast, there was less effect on OPZ (opsonized zymosan)-stimulated TXB2 generation: MZB (IC50 = 1.44 microM) > MZA (IC50 = 3.16 microM) > MZC (IC50 = 3.34 microM) > MZD, MZE and MZF (IC50 > 10 microM). Similarly, PMA-stimulated O2- generation was affected differentially as follows: MZD (apparent IC50 < 0.1 microM) > MZA (IC50 = 0.1 microM) > MZB (IC50 = 3.16 microM) > MZC (IC50 = 3.43 microM) > MZE and MZF (IC50 > 10 microM). In contrast, OPZ-stimulated O2- generation was minimally affected: MZB (IC50 = 4.17 microM) > MZC (IC50 = 9.3 microM) > MZA, MZD, MZE and MZF (IC50 > 10 microM). From the structure-activity relationship perspective, contributing factors to the observed differential bioactivity on TXB2 and O2- generation are the solubility or ionic forms of MZA and D as well as changes such as saturation or oxidation of the beta carboline or 8-membered amine ring. In contrast, the fused 13-membered macrocyclic and isoquinoline ring system, and any substitutions in these rings would not appear to be factors contributing to bioactivity. CONCLUSION: To our knowledge, this is the first experimental study that demonstrates that MZA, at in vitro concentrations that are non toxic to E. coli LPS-activated rat neonatal microglia, potently modulates PMA-stimulated TXB2 and O2- generation. MZA may thus be a lead candidate for the development of novel therapeutic agents for the modulation of TXB2 and O2- release in neuroinflammatory diseases. Marine natural products provide a novel and rich source of chemical diversity that can contribute to the design and development of new and potentially useful anti-inflammatory agents to treat neurodegenerative diseases.

Vitamin C inhibits lipid oxidation in human HDL.

HDL are susceptible to oxidation, which affects their cardioprotective properties. Although several studies have reported inhibition of HDL oxidation by vitamin E, none has determined the potential protective effect of vitamin C, another important blood antioxidant. We investigated whether vitamin C protects HDL from oxidation by incubating HDL (0.2 g of protein/L) at 37 degrees C with cupric (Cu2+) ions (10 micromol/L) in the absence (control) or presence of vitamin C (20-200 micromol/L). In the absence of vitamin C, lipid oxidation in HDL began immediately and proceeded rapidly. Cholesteryl linoleate declined to a minimum, whereas lipid oxidation products (lipid dienes and TBARS) increased to near-maximal levels within 1 h. Vitamin C (50-200 micromol/L) retarded initiation of lipid oxidation for at least 4 h under the same conditions. The ability of vitamin C to preserve the cardioprotective antioxidant function of HDL was also assessed. HDL (0.5 g of protein/L) preincubated with Cu2+ (10 micromol/L) for 2 h in the absence of vitamin C lost antioxidant activity (45.4 +/- 6.2% inhibition of LDL oxidation compared with 93.2 +/- 3.6% for native HDL, P < 0.05). The addition of vitamin C (50-200 micromol/L) during preincubation of HDL with Cu2+, however, resulted in no significant loss of HDL antioxidant activity (77.3 +/- 0.3 to 89.8 +/- 5.4% inhibition of LDL oxidation, P > 0.05 compared with native HDL). Our results demonstrate that vitamin C inhibits lipid oxidation in HDL and preserves the antioxidant activity associated with this lipoprotein fraction.

Cadmium alters the localization of N-cadherin, E-cadherin, and beta-catenin in the proximal tubule epithelium.

Recent studies on proximal tubule-derived cells in culture have shown that Cd has relatively specific damaging effects on the cadherin-dependent junctions between the cells. The objective of the present study was to determine whether Cd can affect cadherin-dependent junctions in the proximal tubule epithelium in vivo. Male Sprague-Dawley rats received subcutaneous injections of Cd (0.6 mg/kg in isotonic saline, 5 days per week for up to 6 weeks). One day each week, 24-h urine samples were collected and analyzed for protein and creatinine. After 5-6 weeks, the Cd-treated animals developed significant proteinuria, with no change in creatinine excretion. Visualization of pan-cadherin immunoreactive materials by immunoperoxidase labeling showed that Cd caused a marked reduction in the intensity of cadherin labeling associated with the apical and the basolateral surfaces of the epithelial cells of the proximal tubule, but no change in the pattern of cadherin labeling in other segments of the nephron. Results of studies utilizing specific antibodies against N-cadherin, E-cadherin, and beta-catenin showed changes in the localization of all three molecules in the proximal tubule. Assessment of cell membrane integrity with trypan blue and ethidium homodimer showed no overt evidence of death in the proximal tubule epithelial cells. Additional results showed that Cd caused only a slight increase in the total levels of glutathione and no significant peroxidation of membrane lipids, indicating only a modest level of oxidative stress. These results indicate that Cd can disrupt cadherin-dependent cell-cell junctions in the proximal tubule, and they raise the possibility that a loss of cadherin-mediated adhesion may contribute to the nephrotoxic effects of Cd.

Vitamin C protects low-density lipoprotein from homocysteine-mediated oxidation.

Homocysteine, an atherogenic amino acid, promotes iron-dependent oxidation of low-density lipoprotein (LDL). We investigated whether vitamin C, a physiological antioxidant, could protect LDL from homocysteine-mediated oxidation. LDL (0.2 mg of protein/ml) was incubated at 37 degrees C with homocysteine (1000 microM) and ferric iron (10-100 microM) in either the absence (control) or presence of vitamin C (5-250 microM). Under these conditions, vitamin C protected LDL from oxidation as evidenced by an increased lag time preceding lipid diene formation (> or = 5 vs. 2.5 h for control), decreased thiobarbituric acid-reactive substances accumulation (< or = 19 +/- 1 nmol/mg when vitamin C > or = 10 microM vs. 32 +/- 3 nmol/mg for control, p <.01), and decreased lipoprotein anodic electrophoretic mobility. Near-maximal protection was observed at vitamin C concentrations similar to those in human blood (50-100 microM); also, some protection was observed even at low concentrations (5-10 microM). This effect resulted neither from altered iron redox chemistry nor enhanced recycling of vitamin E in LDL. Instead, similar to previous reports for copper-dependent LDL oxidation, we found that vitamin C protected LDL from homocysteine-mediated oxidation through covalent lipoprotein modification involving dehydroascorbic acid. Protection of LDL from homocysteine-mediated oxidation by vitamin C may have implications for the prevention of cardiovascular disease.

Assessment of student pharmacists' knowledge concerning folic acid and prevention of birth defects demonstrates a need for further education.

Adequate periconceptional consumption of folic acid can prevent neural tube birth defects, and all women capable of becoming pregnant are recommended to consume 400 microg/d. Most women, however, are unaware of this recommendation and do not consume adequate amounts of folic acid. It is important, therefore, that healthcare professionals, such as pharmacists, be capable of educating women regarding folic acid. The aim of this study was to assess knowledge regarding prevention of birth defects by folic acid among student (future) pharmacists in the final year of a professional degree program. Over a 3-y period (1998-2000), students (n = 98) enrolled in a PharmD program completed a survey consisting of five multiple-choice questions concerning folic acid and birth defects. Almost all students (93.9%) correctly identified folic acid as preventing birth defects. Of these students, many also knew that supplementation should begin before pregnancy (73.9%). Fewer, however, were able to correctly identify either the recommended level of intake (55.4%) or good sources of folic acid (57.6-65.2%). These results show that although student (future) pharmacists are aware of folic acid's ability to prevent birth defects, many lack the specific knowledge needed to effectively counsel women in future clinical practice.