Kelly Karpa

Dopamine D2 and D3 receptors are linked to the actin cytoskeleton via interaction with filamin A

We have used a yeast two-hybrid approach to uncover protein interactions involving the D2-like subfamily of dopamine receptors. Using the third intracellular loop of the D2S and D3 dopamine receptors as bait to screen a human brain cDNA library, we identified filamin A (FLN-A) as a protein that interacts with both the D2 and D3 subtypes. The interaction with FLN-A was specific for the D2 and D3 receptors and was independently confirmed in pull-down and coimmunoprecipitation experiments. Deletion mapping localized the dopamine receptor–FLN-A interaction to the N-terminal segment of the D2 and D3 dopamine receptors and to repeat 19 of FLN-A. In cultures of dissociated rat striatum, FLN-A and D2 receptors colocalized throughout neuronal somata and processes as well as in astrocytes. Expression of D2 dopamine receptors in FLN-A-deficient M2 melanoma cells resulted in predominant intracellular localization of the D2 receptors, whereas in FLN-A-reconstituted cells, the D2 receptor was predominantly localized at the plasma membrane. These results suggest that FLN-A may be required for proper cell surface expression of the D2 dopamine receptors. Association of D2 and D3 dopamine receptors with FLN-A provides a mechanism whereby specific dopamine receptor subtypes may be functionally linked to downstream signaling components via the actin cytoskeleton.

The Pharmacists’ Role in the Patient-Centered Medical Home (PCMH) A white paper created by the Health Policy Committee of the Pennsylvania Pharmacists Association (PPA)

Thefollowing are contributors to the writing of this paper (in alphabetical order): Hildegarde Berdine, PharmD, BCPS,CDE Associate Professor ofPharmacy Practice, Duquesne University, Mylan School of Pharmacy; Tanya Dougherty, PharmD, BCPS, Clinical Pharmacy Coordinator, Penn Presbyterian Medical Center; Jonathan Ference, PharmD, BCPS, Associate Professor, Wilkes University School ofPharmacy and Nursing; Kelly Karpa, PhD, RPh, Director, Medical Pharmacology Instruction, Penn State University College ofMedicine; Jacqueline Klootwyk, PharmD, BCPS, Assistant Professor, Thomas Jefferson University School ofPharmacy; Melinda Kozminski, PharmD, BCACP, Clinical Pharmacist, Gateway Health Plan; Nicholas Leon, PharmD, BCPS, BCACP, Thomas Jefferson University School of Pharmacy; Maria Osborne, PharmD, Clinical Pharmacist-Family Practice, UPMC St. Margaret; Adam C. Welch, PharmD, MBA, BCACP, Associate Professor, Wilkes University School of Pharmacy and Nursing; and Vincent J. Willey, PharmD, Associate Professor and Vice Chair, Department of Pharmacy Practice and Pharmacy Administration, Philadelphia College of Pharmacy, University of the Sciences.

Probiotics for Clostridium Difficile Diarrhea: Putting it into Perspective

Clostridium difficile diarrhea is an expensive, life-threatening infection associated with serious morbidity and mortality, even among previously healthy individuals. Relapses from the infection are common following standard antibiotic treatments, with 3–5% of patients who contract C. difficile diarrhea unable to discontinue vancomycin due to continual relapses. Such patients may have a focal immunodeficiency in which they tail to mount an immune response against C. difficile. For these individuals, antimicrobial therapies are unable to eradicate the microorganism because no antibiotics are capable of killing C. difficile spores. Although they are considered alternative medicine, probiotics have provided a safe and effective means of restoring gastrointestinal flora and alleviating diarrhea, particularly for individuals experiencing multiple relapses.

Bacillus Coagulans GBI-30 (BC30) improves indices of Clostridium difficile-Induced colitis in mice

BackgroundProbiotics have beneficial effects in rodent models of Clostridium difficile (C. diffiicle)-induced colitis. The spore forming probiotic strain Bacillus Coagulans GBI-30, 6086 (BC30) has demonstrated anti-inflammatory and immune-modulating effects in vitro. Our goal was to determine if BC30 improved C. difficile-induced colitis in mice. Starting on study day 0, female C57BL/6 mice were dosed by oro-gastric gavage for 15 days with vehicle (saline) or BC30 (2 × 109 CFU per day). Mice in the C. difficile groups received an antibiotic mixture (study days 5 to 8 in the drinking water), and clindamycin (10 mg/kg, i.p., on study day 10). The C. difficile strain VPI 10463 was given by gavage at 104 CFU to induce colitis on day 11. On day 16, stools and colons were collected for further analyses.ResultsAll mice treated with BC30 survived on study day 13, while two mice treated with vehicle did not survive. On day 12, a significant difference (p = 0.0002) in the percentage of mice with normal stools (66.7%) was found in the BC30/C. difficile group, as compared to the vehicle/C. diffcile group (13.0%). On study day 16, 23.8% of mice treated with BC30 had normal stools, while this value was 0% with vehicle treatment (p value = 0.0187). On this day, the stool consistency score for the BC30/C. difficile group (1.1 ± 0.2) was significantly lower (p < 0.05) than for the vehicle/C. difficile cohort (1.9 ± 0.2). BC30 modestly attenuated the colonic pathology (crypt damage, edema, leukocyte influx) that was present following C. difficile infection. Colonic MIP-2 chemokine contents (pg/2 cm colon) were: 10.2 ± 0.5 (vehicle/no C. difficile), 24.6 ± 9.5 (vehicle/C. difficile) and 16.3 ± 4.3 (BC30/C. difficle).ConclusionThe probiotic BC30 improved some parameters of C. difficile-induced colitis in mice. BC30 prolonged the survival of C. diffiicle infected mice. Particularly, this probiotic improved the stool consistency of mice, in this infectious colitis model.

A retrospective chart review to identify perinatal factors associated with food allergies

BackgroundGut flora are important immunomodulators that may be disrupted in individuals with atopic conditions. Probiotic bacteria have been suggested as therapeutic modalities to mitigate or prevent food allergic manifestations. We wished to investigate whether perinatal factors known to disrupt gut flora increase the risk of IgE-mediated food allergies.MethodsBirth records obtained from 192 healthy children and 99 children diagnosed with food allergies were reviewed retrospectively. Data pertaining to delivery method, perinatal antibiotic exposure, neonatal nursery environment, and maternal variables were recorded. Logistic regression analysis was used to assess the association between variables of interest and subsequent food allergy diagnosis.ResultsRetrospective investigation did not find perinatal antibiotics, NICU admission, or cesarean section to be associated with increased risk of food allergy diagnosis. However, associations between food allergy diagnosis and male gender (66 vs. 33; p=0.02) were apparent in this cohort. Additionally, increasing maternal age at delivery was significantly associated with food allergy diagnosis during childhood (OR, 1.05; 95% CI, 1.017 to 1.105; p=0.005).ConclusionsGut flora are potent immunomodulators, but their overall contribution to immune maturation remains to be elucidated. Additional understanding of the interplay between immunologic, genetic, and environmental factors underlying food allergy development need to be clarified before probiotic therapeutic interventions can routinely be recommended for prevention or mitigation of food allergies. Such interventions may be well-suited in male infants and in infants born to older mothers.

Bacillus coagulans GBI-30, 6086 limits the recurrence of Clostridium difficile-Induced colitis following vancomycin withdrawal in mice

BackgroundRecently, we found that the probiotic strain Bacillus coagulans GBI-30, 6086 (GanedenBC30) improved indices of Clostridium difficile (C. difficile)-induced colitis in mice (Fitzpatrick et al., Gut Pathogens, 2011). Our goal was to determine if BC30 could also prevent the recurrence of C. difficile-induced colitis in mice, following initial treatment with vancomycin. During study days 0 through 5, mice were treated with antibiotics. On day 6, the C. difficile strain VPI 10463 was given by oro-gastric gavage at ≈ 5x104 CFU to induce colitis. Mice were treated on study days 6 to 10 with vancomycin (50 mg/kg) (vanco) or vehicle (saline) by gavage. On days 10 to16, mice were dosed by gavage with saline vehicle or BC30 (2 x 109 CFU per day). Mice were monitored for mortality, weight loss and diarrhea. On study days 14, 16 and 17, stools and colons were collected for analyzing other parameters of colitis.ResultsThe mean stool consistency score in Vehicle/C.difficile/Vanco mice increased from 0.4 (day 10) to a range of 1.1 to 1.4 (days 14 to 17), indicating the recurrence of colitis. On days 13 through 17, the stool consistency scores for the vancomycin/BC30 mice were significantly lower (p< 0.05) than for the vancomycin/vehicle cohort of animals. On day 17, 88.9% of mice treated with BC30 had normal stools, while this value was 0% with vehicle treatment (p value = 0.0004). Colonic myeloperoxidase (Units/2 cm colon) was significantly (p < 0.05) reduced from 4.3 ± 0.7 (Vehicle/C.difficile/Vanco) to 2.6 ± 0.2 (BC30/C. Difficle/Vanco). The colonic histology score and Keratinocyte derived-chemokine level in the colon were also lower in BC30 treated mice.SummaryIn BC30-treated mice, there was evidence of better stool consistency, as well as improved biochemical and histological indices of colitis, following initial treatment of animals with vancomycin.ConclusionBC30 limited the recurrence of CD-induced colitis following vancomycin withdrawal in mice.

Creating a Virtual Pharmacology Curriculum in a Problem-Based Learning Environment: One Medical School's Experience

Integrating pharmacology education into a problem-based learning (PBL) curriculum has proven challenging for many medical schools, including the Pennsylvania State University College of Medicine (Penn State COM). In response to pharmacology content gaps in its PBL-intensive curriculum, Penn State COM in 2003 hired a director of medical pharmacology instruction to oversee efforts to improve the structure of pharmacology education in the absence of a stand-alone course. In this article, the authors describe the ongoing development of the virtual pharmacology curriculum, which weaves pharmacology instruction through the entire medical school curriculum with particular emphasis on the organ-based second year. Pharmacology is taught in a spiraling manner designed to add to and build upon students’ knowledge and competency. Key aspects of the virtual curriculum (as of 2011) include clearly stated and behaviorally oriented pharmacology learning objectives, pharmacology study guides that correspond to each PBL case, pharmacology review sessions that feature discussions of United States Medical Licensing Examination (USMLE)-type questions, and pharmacology questions for each PBL case on course examinations to increase student accountability. The authors report a trend toward improved USMLE Step 1 scores since these initiatives were introduced. Furthermore, graduates’ ratings of their pharmacology education have improved on the Medical School Graduation Questionnaire. The authors suggest that the initiatives they describe for enhancing pharmacology medical education are relevant to other medical schools that are also seeking ways to better integrate pharmacology into PBL-based curricula.

Development and implementation of an herbal and natural product elective in undergraduate medical education

BackgroundMedical students have consistently expressed interest in learning about alternative healing modalities, especially herbal and natural products. To fill this void in medical education at our institution, a novel elective was developed and implemented for fourth year medical students. This herbal/natural product course uses guest lecturers, classroom presentations, and active learning mechanisms that include experiential rotations, case-based learning, and team-based learning to increase student knowledge of herbal/natural product safety and efficacy.MethodsKnowledge outcomes were evaluated via administration of a pre- and post-course test (paired student t-test). End-of-course evaluations (Likert-type questions and narrative responses) were used to assess student opinion of knowledge and skills imparted by the elective and overall course content (mean, standard deviation).ResultsOver three academic years, 23 students have enrolled in this elective. More than 60% of participants have been female and nearly half of the students (43%) have pursued residencies in primary care. Completion of the course significantly increased student knowledge of common herbal/natural product mechanisms, uses, adverse effects, and drug-interactions as determined by a pre- and post-course knowledge assessment (45%u2009±u200910% versus 78%u2009±u20096%; pu2009<u20090.0001). The course was highly rated by enrollees (overall course quality, 4.6 of 5.0u2009±u20090.48) who appreciated the variety of activities to which they were exposed and the open classroom discussions that resulted. While students tended to view some alternative medical systems with skepticism, they still believed it was valuable to learn what these modalities encompass.ConclusionsDevelopment and implementation of a herbal/natural product elective that engages undergraduate medical students through active learning mechanisms and critical analysis of the literature has proven effective in increasing knowledge outcomes and is deemed to be a valuable curricular addition by student participants. In the future, it will be of interest to explore mechanisms for expanding the course to reach a larger number of students within the time, financial, and logistical constraints that currently exist.

Medication safety curriculum: enhancing skills and changing behaviors

BackgroundAdverse drug reactions are a leading cause of death in the United States. Safe and effective management of complex medication regimens is a skill for which recent medical school graduates may be unprepared when they transition to residency. We wished to assess the impact of a medication safety curriculum on student competency when evaluating medication therapeutic appropriateness as well as evaluate students’ ability to transfer curricular material to management of patients in clinical settings.MethodsTo prepare 3rd and 4th year medical students to critically evaluate medication safety and appropriateness, we developed a medication reconciliation/optimization curriculum and embedded it within a Patient-Centered Medical Home longitudinal elective. This curriculum is comprised of a medication reconciliation workshop, in-class and individual case-based assignments, and authentic patient encounters in which medication management skills are practiced and refined. Pre- and post-course competency and skills with medication reconciliation/optimization are evaluated by assessing student ability to identify and resolve medication-related problems (MRPs) in case-based assignments using paired difference tests. A group of students who had wished to enroll in the elective but whose schedule did not permit it, served as a comparison group.ResultsStudents completing the curriculum (nu2009=u200945) identified 75xa0% more MRPs in case assignments compared to baseline. No changes from baseline were apparent in the comparison group. Enrolled students were able to transfer their skills to the care of authentic patients; these students identified an average of 2.5 MRPs per patient from a panel of individuals that had recently transitioned from hospital to home. Moreover, patient questionnaires (before and several months following the medication encounters with assigned students) indicated that patients felt more knowledgeable about several medication parameters as a result of the student-led medication encounter. Patients also indicated that students helped them overcome barriers to medication adherence (e.g. cost, transportation, side effects).ConclusionsNovice learners may have difficulty transitioning from knowledge of basic pharmacology facts to application of that information in clinical practice. Our curriculum appears to bridge that gap in ways that may positively impact patient care.

Patient experience and use of probiotics in community-based health care settings.

Objective To investigate patient experience with probiotics and factors that influence probiotic use among adult patients. Method Patients were invited to complete a questionnaire that assessed their experiences and opinions regarding probiotics. Questionnaires were distributed to patients seeking primary health care services at a family and community medicine practice site and a community pharmacy. Patients were invited to complete the questionnaire while awaiting the physician or waiting for prescriptions to be filled. Results Overall, 162 surveys were completed and returned (66% response rate) from patients aged 18 to 89 years of age (mean 49.5 years). Most patients (n=107; 65%) were familiar with the term “probiotic”, and 49 patients (29.9%) had personally used the supplements in the past. Of those who had used probiotics, the majority (57%) had used the supplements to maintain “good gastrointestinal health” and most (59%) felt that the supplements had been beneficial. However, most (59%) had not informed their health care provider about their use of the supplements. Conclusion Use of probiotic supplements is common among consumers, but may not be reported to health care providers.