A Culinary Medicine Elective Course Incorporating Lifestyle Medicine for Medical Students.

OBJECTIVE: The purpose of this study is to describe our culinary medicine elective course with a lifestyle modification focus and to evaluate the students' perceived knowledge and attitudes in lifestyle medicine. METHODS: Pre- and post-surveys including quantitative assessment, Likert-type questions, and one open-ended response question to assess students' perceived knowledge of nutrition and lifestyle medicine were distributed to osteopathic medical students who participated in the culinary medicine elective course. The Mann-Whitney U test and dependent t test were used where appropriate based on normality. RESULTS: Compared to the pre-course survey, students who responded "strongly agree" in questions related to nutrition counseling in the post-course survey were 26.5 to 31.3% higher (p < 0.05). Based on the post-course survey (n = 34), 33 students responded either "strongly agree" (n = 25, 73.5%) or "agree" (n = 8, 23.5%) to the question of "increased my knowledge of nutrition." CONCLUSIONS: Culinary medicine courses with a lifestyle medicine focus may be effective in increasing medical students' confidence and perceived knowledge of nutrition and lifestyle medicine.

Pedigree Analysis: A Team-Based Learning Activity.

INTRODUCTION: Regardless of their specialty, physicians, particularly those practicing primary care in rural areas, typically encounter genetic conditions. Therefore, it is important to incorporate genetic principles into medical training prior to students' clinical rotations. METHODS: The advance preparation assignment for this team-based learning (TBL) resource includes lectures and directed study assignments on the following topics: Mendelian genetics, sexual genetics, population genetics, and pedigree analysis. Students then demonstrate their understanding of the content through a formal TBL lesson utilizing a gallery walk for group application exercises. RESULTS: Course evaluations clearly indicated that students enjoyed and learned from this TBL. Student performance on the team readiness assurance test and the summative exam scores showed significant signs of improvement when compared to individual readiness assurance test performance. DISCUSSION: In this TBL module, students develop an understanding of basic genetics, pedigree analysis, and calculation of risks for inheriting autosomal recessive and X-linked recessive diseases. This TBL can be easily adapted to other allied health programs.

Community adaptations to an impending food desert in rural Appalachia, USA.

INTRODUCTION: The United States Department of Agriculture (USDA) describes a food desert as an urban neighborhood or rural town without ready access to fresh, healthy, and affordable food. An estimated 2.3 million rural Americans live in food deserts. One goal of the USDA is to eliminate food deserts. However, at a time when some food deserts are being eliminated, hundreds of grocery stores are closing, causing other food deserts to arise. The literature is scarce on how a community adapts to an impending food desert. Alderson, West Virginia, USA (population 1184) rallied to face an impending food desert when the only grocery store in town closed in December 2014. This study investigated how this small rural community adapted to its oncoming food desert. METHODS: A community member survey was administered to 155 Alderson families (49%) to determine how the new food desert affected family food acquisition and storage behaviors. A restaurant survey was given to the town's four restaurants to determine how the food desert affected their businesses. Sales data for a new food hub (Green Grocer) was obtained to see if this new initiative offset the negative effects of the food desert. ANOVA and t-tests were used to compare group numerical data. Two group response rates were compared by testing the equality of two proportions. Categorical data were analyzed with the χ2 or frequency distribution analysis. Group averages are reported as mean ± standard error of the mean. Significance for all analyses was set at p<0.05. RESULTS: Even though 86% of the population shopped at the new Green Grocer, 77% did most of their shopping at a store at least 17.7 km (11 miles) from home. The number of long-distance monthly shopping trips made after the food desert (3.3±0.4) did not change significantly (p=0.16) from the number before the food desert (2.8±0.3). Price comparisons among the Green Grocer and three distant supermarkets showed a 30% savings by traveling to distant supermarkets. Frequency of monthly restaurant visits did not change after the emergence of the food desert (2.98±0.54 vs 3.05±0.51, p=0.85). However, restaurant patrons requested to buy fresh produce and dairy from the restaurants to use for their own home cooking. Food pantry use increased by 43%, with community members requesting more fresh produce, meat, and dairy. The food desert triggered a 21% increase in home gardening and an 11% increase in home food preservation. CONCLUSIONS: Opening a Green Grocer offset only some of the effects of the food desert, because community members use it as a convenience store to purchase fresh produce and dairy products that families may lack before their next long-distance trip to a supermarket. Alderson's low-income residents now rely more heavily on food pantry assistance, while a small number of other residents have started gardening and food preservation. The first factor governing food acquisition behavior in rural Appalachia is food pricing, with the proximity of food access coming in second. How to overcome these two major barriers to food security in the midst of current economics and marketing remains to be answered.

Misconceptions about health and disease prevention behaviors of rural Appalachian Americans.

BACKGROUND: Appalachia is one of the unhealthiest regions in the United States due to poor disease prevention behaviors. OBJECTIVE: Determine if self-perceived health of rural Appalachians is related to participation in disease prevention behaviors. METHODS: Rural Appalachian adults (n=437) were surveyed regarding their self-perceived health and disease prevention behaviors. Healthy behaviors included: moderate (≥ 90 min/wk) and vigorous (≥ 45 min/wk) physical activity, sugared drink consumption (≤ 1 sugared drink/d), smoking (non-smoker), alcohol consumption (≤ 1 drink/d), blood pressure (≤ 120/80 mm Hg), and fast food consumption (≤ 1 time/wk). Participants were grouped where healthy = (self-health rating > 5 on a 0-10 scale), BMI < 25, and blood pressure (≤ 120/80 mm Hg). Jaccard Binary Similarity (JBS) coefficients and Russell and Rao (RR) dichotomy coefficients determined the association and predictability of self-health ratings and disease prevention behaviors. T-tests determined group differences in the number of disease prevention behaviors. RESULTS: Individuals who reported being healthy had high JBS coefficients for having healthy sugared drink consumption (0.552), not smoking (0.704), low alcohol consumption (0.742), and low fast food consumption (0.481). RR results were similar to JBS results. Not smoking and low alcohol consumption were highly correlated (r=0.87). Those with a good health perception practiced more disease prevention behaviors (mean±SEM, 2.84±0.06) than those with a poor health perception (2.19±0.10, p<0.001). Good health perceptions were not strongly related to obesity and inactivity. CONCLUSIONS: Appalachians are not indifferent about their health. However, Appalachians may not understand how inactivity and obesity relate to disease.

Tobacco dependence curricula in US osteopathic medical schools: a follow-up study.

CONTEXT: Tobacco use is the leading preventable cause of illness and death in the United States. A 1998 survey of US osteopathic medical schools identified deficiencies in tobacco dependence curricula. OBJECTIVE: To assess the current content and extent of tobacco dependence education and intervention skills in US osteopathic medical school curricula. DESIGN: An electronic survey. SETTING: Osteopathic medical schools with students enrolled for the 2009-2010 academic year. PARTICIPANTS: Twenty-seven osteopathic medical school deans or their designated administrators. MAIN OUTCOME MEASURES: Reported instruction in 7 basic science and 6 clinical science content areas (elective or required) and hours of tobacco dependence education were assessed and compared with the 1998 data. RESULTS: The mean (standard deviation) number of content areas reported as covered in 2010 was 10.6 (2.3) (6.1 [1.2] basic science areas, 4.6 [1.3] clinical science areas). Seventeen of 27 respondents (63%) reported that smokeless tobacco content was covered at their school, and 9 of 27 (33%) reported that the stages of change counseling technique was covered. Compared with 1998, a significant increase was noted in the percentage of schools covering tobacco dependence (92.6% in 2010 compared with 57.9% in 1998, P=.0002). Reported hours of tobacco dependence instruction were also significantly higher in 2010 compared with those in 1998 (Fisher exact test, P<.05). No statistically significant changes were found in the proportion of schools covering all 13 content areas (15.7% vs 22.2%), the proportion covering motivational interviewing in detail (26.3% vs 33.3%), or the proportion requiring curricula on smokeless tobacco (57.9% vs 59.3%). CONCLUSION: Osteopathic medical school respondents reported more instruction on tobacco dependence in 2010 compared with those in 1998. However, some important basic science and clinical science content areas are not being adequately taught in US osteopathic medical schools.

Self-rated health in rural Appalachia: health perceptions are incongruent with health status and health behaviors.

BACKGROUND: Appalachia is characterized by poor health behaviors, poor health status, and health disparities. Recent interventions have not demonstrated much success in improving health status or reducing health disparities in the Appalachian region. Since one's perception of personal health precedes his or her health behaviors, the purpose of this project was to evaluate the self-rated health of Appalachian adults in relation to objective health status and current health behaviors. METHODS: Appalachian adults (n = 1,576) were surveyed regarding health behaviors - soda consumer (drink ≥ 355 ml/d), or non-consumer (drink < 355 ml/d), fast food consumer (eating fast food ≥ 3 times/wk) or healthy food consumer (eating fast food < 3 times/wk), smoking (smoker or non-smoker), exercise (exerciser > 30 min > 1 d/wk) and sedentary (exercise < 30 min 1 d/wk), blood pressure medication (yes, no), and self-rated health. Blood pressure was measured through auscultation and serum cholesterol measured via needle prick. Weight status was based on BMI: normal weight (NW ≥ 18.5 and < 25.0), overweight (OW ≥ 25.0 and < 30.0), and obese (OB ≥ 30.0). Jaccard Binary Similarity coefficients, odds ratios, chi-square, and prevalence ratios were calculated to evaluate the relationships among self-rated health, objective health status, and health behaviors. Significance was set at p < 0.05. RESULTS: Respondents reported being healthy, while being sedentary (65%), hypertensive (76%), overweight (73%), or hyperlipidemic (79%). Between 57% and 66% of the respondents who considered themselves healthy had at least two disease conditions or poor health behaviors. Jaccard Binary Similarity coefficients and odds ratios showed the probability of reporting being healthy when having a disease condition or poor health behavior was high. CONCLUSIONS: The association between self-rated health and poor health indicators in Appalachian adults is distorted. The public health challenge is to formulate messages and programs about health and health needs which take into account the current distortion about health in Appalachia and the cultural context in which this distortion was shaped.

A novel Bcl-2 small molecule inhibitor 4-(3-methoxy-phenylsulfannyl)-7-nitro-benzofurazan-3-oxide (MNB)-induced apoptosis in leukemia cells.

A novel small molecule inhibitor, 4-(3-methoxy-phenylsulfannyl)-7-nitro-benzofurazan-3-oxide (MNB), competes with the Bak BH3 peptide to bind Bcl-2 protein with a binding affinity of IC(50) = 0.70 microM, as assessed by a fluorescence polarization based binding assay. HL-60 cells express the highest levels of Bcl-2 among the cell lines examined. Treated with 5 microM of MNB only for 6 h, 85% of HL-60 cells were detected to undergo apoptosis. Pan-caspase inhibitor, Z-VAD-FMK, blocks MNB-induced apoptosis in HL-60 cells. Caspase-2, caspase-3, caspase-7, caspase-8, caspase-9, and PARP activation were observed at as early as 4 to 6 h of MNB treatment. In addition, it has been confirmed that the caspase-3 specific inhibitor, Z-DEVD-FMK, blocks the activation of caspase-8 in MNB-treated HL-60 cells. MNB treatment does not change Bcl-2 or Bax expression level in HL-60 cells, but causes Bid cleavage. Further experiments have illustrated that MNB inhibits the heterodimerization of Bcl-2 with Bax or Bid, reduces the mitochondrial membrane potential (DeltaPsimt), and induces cytochrome c release from mitochondria in HL-60 cells. These results suggest that MNB induces apoptosis in HL-60 by inhibiting the heterodimerization of Bcl-2 with pro-apoptosis Bcl-2 members, resulting in a decrease in the mitochondrial membrane potential and cytochrome c release, activation of caspases and PARP; it is a caspase-dependent process in which the activation of caspase-8 is dependent on the mitochondrial apoptosis signal transduction pathway. MNB prolongs the life spans of HL-60 bearing mice, potently kills fresh AML and ALL cells, indicating that it has the potential to be developed to treat leukemia.

Bioinformatics-based discovery and characterization of an AKT-selective inhibitor 9-chloro-2-methylellipticinium acetate (CMEP) in breast cancer cells.

AKT is a promising target for anticancer drug development. In this work, a bioinformatics approach was applied to search for AKT inhibitors based on the correlation analysis between phospho-Ser473 AKT expression level and the antiproliferative data of NCI small molecule compounds against NCI 60 cancer cell lines, the candidate compounds were then subject to AKT kinase assay. The possible effects of potent compound on PI3K/AKT, PDK1, and MAPK, its antiproliferative and apoptosis-inducing effects on breast cancer cells which have high-levels of AKT activation were assessed by Western blot analysis, cell viability assay, and apoptosis assay. One compound, CMEP (NSC632855, 9-chloro-2-methylellipticinium acetate) was identified with all three correlation algorithm, Pearson's, Sperman's, and Kendall's, showing a high-ranked correlation coefficient. CMEP inhibits only AKT, but does not inhibit PI3K, PDK1, or MAPK. CMEP also inhibits heregulin-induced AKT activation, does not inhibit heregulin-induced MAPK activation in MCF-7 breast cancer cells. Increased concentrations of ATP reverse the AKT inhibitory effect of CMEP. CMEP inhibits growth and induces apoptosis in breast cancer cells which have high-levels of AKT activation and lack functional PTEN; however, CMEP only shows a minimal activity in NIH3T3 cells which do not have AKT activation. In conclusion, a lead compound CMEP, as an AKT selective inhibitor has been identified started with a bioinformatics-based approach. CMEP inhibits growth and induces apoptosis in cancer cells which have high-levels of AKT activation and lack PTEN or harbor PTEN mutation.

Gossypol induces apoptosis in human PC-3 prostate cancer cells by modulating caspase-dependent and caspase-independent cell death pathways.

The rate of gossypol-induced apoptosis does not correlate very well with the same dose of gossypol-induced cell growth inhibition, indicating an anti-proliferative effect of gossypol. Using a co-immunoprecipitation assay, it was observed that the level of Bcl-X(L) protein bound to Bax was clearly lower than that of Bcl-2 protein at 5 micro M of gossypol treatment, and the level of Bim protein bound to Bcl-X(L) was lowered at 20 micro M of gossypol treatment for 24 h, implicating that gossypol inhibits the heterodimerization of Bcl-X(L) with Bax and Bim. Gossypol-induced apoptosis is partly suppressed by as low as 0.5 micro M, but not abolished by as high as 50 micro M of a broad range caspase inhibitor, Z-VAD-FMK, suggesting that gossypol-induced apoptosis is both caspase-dependent and -independent. Furthermore, the release of apoptosis inducing factor (AIF), which triggers caspase-independent apoptosis, from mitochondria to cytosol was observed in PC-3 cells exposed to gossypol treatment. In conclusion, gossypol inhibits the proliferation and induces apoptosis in PC-3 cells. Gossypol-induced apoptosis is, at least, through inhibiting the heterodimerization of Bcl-X(L)/Bcl-2 with pro-apoptosis molecules, followed by a caspase-dependent and -independent process which involves the release of AIF from the mitochondria to cytosol.

Identification of hnRNPs K, L and A2/B1 as candidate proteins involved in the nutritional regulation of mRNA splicing.

Nutrient regulation of glucose-6-phosphate dehydrogenase (G6PD) expression occurs through changes in the rate of splicing of G6PD pre-mRNA. This posttranscriptional mechanism accounts for the 12- to 15-fold increase in G6PD expression in livers of mice that were starved and then refed a high-carbohydrate diet. Regulation of G6PD pre-mRNA splicing requires a cis-acting element in exon 12 of the pre-mRNA. Using RNA probes to exon 12 and nuclear extracts from livers of mice that were starved or refed, proteins of 60 kDa and 37 kDa were detected bound to nucleotides 65-79 of exon 12 and this binding was decreased by 50% with nuclear extracts from refed mice. The proteins were identified as hnRNPs K, L, and A2/B1 by LC-MS/MS. The decrease in binding of these proteins to exon 12 during refeeding was not accompanied by a decrease in the total amount of these proteins in total nuclear extract. HnRNPs K, L and A2/B1 have known roles in the regulation of mRNA splicing. The decrease in binding of these proteins during treatments that increase G6PD expression is consistent with a role for these proteins in the inhibition of G6PD mRNA splicing.

An exonic splicing silencer is involved in the regulated splicing of glucose 6-phosphate dehydrogenase mRNA.

The inhibition of glucose-6-phosphate dehydrogenase (G6PD) expression by arachidonic acid occurs by changes in the rate of pre-mRNA splicing. Here, we have identified a cis-acting RNA element required for regulated splicing of G6PD mRNA. Using transfection of G6PD RNA reporter constructs into rat hepatocytes, the cis-acting RNA element involved in this regulation was localized to nucleotides 43-72 of exon 12 in the G6PD mRNA. In in vitro splicing assays, RNA substrates containing exon 12 were not spliced. In contrast, RNA substrates containing other regions (exons 8 and 9 or exons 10 and 11) of the G6PD mRNA were efficiently spliced. Furthermore, exon 12 can inhibit splicing when substituted for other exons in RNA substrates that are readily spliced. This activity of the exon 12 regulatory element suggests that it is an exonic splicing silencer. Consistent with its activity as a splicing silencer, spliceosome assembly was inhibited on RNA substrates containing exon 12 compared with RNAs representing other regions of the G6PD transcript. Elimination of nucleotides 43-72 of exon 12 did not restore splicing of exon 12-containing RNA; thus, the 30-nucleotide element may not be exclusively a silencer. The binding of heterogeneous nuclear ribonucleoproteins K, L, and A2/B1 from both HeLa and hepatocyte nuclear extracts to the element further supports its activity as a silencer. In addition, SR proteins bind to the element, consistent with the presence of enhancer activity within this sequence. Thus, an exonic splicing silencer is involved in the inhibition of splicing of a constitutively spliced exon in the G6PD mRNA.

Improved expression and characterization of Ca2+-ATPase and phospholamban in High-Five cells.

The Ca2+-ATPase accounts for the majority of Ca2+ removed from the cytoplasm during cardiac muscle relaxation. The Ca2+-ATPase is regulated by phospholamban, a 52 amino acid phosphoprotein, which inhibits Ca2+-ATPase activity by decreasing the apparent affinity of the ATPase for Ca2+. To study the physical mechanism of Ca2+-ATPase regulation by phospholamban using spectroscopic and kinetic experiments, large amounts of both proteins are required. Therefore, we developed a Ca2+-ATPase and phospholamban preparation based on the baculovirus-insect cell expression system using High-Five insect cells to produce large amounts of microsomal vesicles that contain either Ca2+-ATPase expressed alone or Ca2+-ATPase co-expressed with phospholamban. The expressed proteins were characterized using immunofluorescence spectroscopy, Ca2+ -ATPase activity assays, Ca2+ uptake and efflux assays, and Western blotting. Our purification method yields 140 mg of microsomal protein per liter of infection (1.7 x 10(9)cells), and the Ca2+-ATPase and phospholamban account for 16 and 1.4%, respectively, of the total microsomal protein by weight, yielding a phospholamban:Ca2+-ATPase ratio of 1.6:1, similar to that observed in native cardiac SR vesicles. The enzymatic properties of the expressed Ca2+-ATPase are also similar to those observed in native cardiac SR vesicles, and when co-expressed with phospholamban, the Ca2+-ATPase is functionally coupled to phospholamban similar to that observed in cardiac SR vesicles.