Introduction of a new method for two-dimensional NMR quantitative analysis in metabolomics studies.

NMR is one of the most important platforms for metabolomic studies. Though 2D NMR has been applied in metabolomics, most applications have mainly focused on metabolite identification whilst limitations causing a bottle-neck for applying high-throughput 2D NMR data for quantity related statistical analysis lies on the data interpretation methods. In this study, instead of using the traditional methods of calculating the 2D NMR data to search for the important features, a new procedure, which applies the high-resolution 1D NMR metabolites chemical shift range to filter the 2D NMR data, was developed. This new method was demonstrated using both a mixture of standard metabolites and a case study on plant extracts using 2D non-uniform sampling (NUS) total correlation spectroscopy (TOCSY) data. As a result, our method successfully filtered out the important features with a high success rate, and the extracted peaks showed high linearity between the calculated intensities and the concentrations of metabolites from a range of 0.05 mM-2 mM. The method was successfully applied to a metabolomics case study which included 18 Begonia samples that showed excellent peak extractions. In summary, our study has provided a practical new 2D NMR data extraction method for use in future metabolomics studies.

Photodegradation of ibuprofen and four other pharmaceutical pollutants on natural pigments sensitized TiO2 nanoparticles.

Pharmaceuticals and personal care products (PPCPs) in water system have drawn increasing concerns in recent years. TiO2 -based photodegradation has shown great potential as a low-cost and sustainable technology in water treatment, however, can only use the UV light range of solar radiation which makes the system less efficient. Dyes have been studied to improve the TiO2 system light-harvesting range, but studies on environmental friendly natural dyes are rare. In this study, a screening method using UV-Vis spectra analysis was carried out on a group of 22 different tropical natural plants for the potential applications on dye-sensitized TiO2 in PPCP treatment. As a result, Begonia "Martin's Mystery" significantly increased TiO2 photodegradation efficiency toward ibuprofen treatments which is first time reported in literature as our best knowledge. Moreover, the promising discovery of Begonia application in ibuprofen treatment has been successfully applied to warfarin and famotidine treatment. Similar results were expanded to many other Begonia species which indicate that Begonia extracts could be excellent sensitizers for TiO2 -based photodegradation of PPCPs. Our discovery suggested that the screening process may potentially open a brand-new way for future TiO2 photodegradation studies before the complex and time-consuming detailed mechanism studies. PRACTITIONER POINTS.: Natural dyes were screened as sensitizers for TiO2 photodegradation of ibuprofen. Ibuprofen photodegradation efficiency was increased twice using Begonia "Martin's Mystery." The Begonia applications were extended to warfarin, trimethoprim, and famotidine. Promising results were also observed using five other Begonia species.

Cytochrome P450 Genetic Variation Associated with Tamoxifen Biotransformation in American Indian and Alaska Native People.

Despite evidence that pharmacogenetics can improve tamoxifen pharmacotherapy, there are few studies with American Indian and Alaska Native (AIAN) people. We examined variation in cytochrome P450 (CYP) genes (CYP2D6, CYP3A4, CYP3A5, and CYP2C9) and tamoxifen biotransformation in AIAN patients with breast cancer (n = 42) from the Southcentral Foundation in Alaska and the Confederated Salish and Kootenai Tribes in Montana. We tested for associations between CYP diplotypes and plasma concentrations of tamoxifen and metabolites. Only the CYP2D6 variation was significantly associated with concentrations of endoxifen (P = 0.0008) and 4-hydroxytamoxifen (P = 0.0074), tamoxifen's principal active metabolites, as well as key metabolic ratios. The CYP2D6 was also the most significant predictor of active metabolites and metabolic ratios in a multivariate regression model, including all four genes as predictors, with minor roles for other CYP genes. In AIAN populations, CYP2D6 is the largest contributor to tamoxifen bioactivation, illustrating the importance of validating pharmacogenetic testing for therapy optimization in an understudied population.

Partnership with the Confederated Salish and Kootenai Tribes: Establishing an Advisory Committee for Pharmacogenetic Research.

BACKGROUND: Inclusion of American Indian and Alaska Native (AI/AN) populations in pharmacogenetic research is key if the benefits of pharmacogenetic testing are to reach these communities. Community-based participatory research (CBPR) offers a model to engage these communities in pharmacogenetics. OBJECTIVES: An academic-community partnership between the University of Montana (UM) and the Confederated Salish and Kootenai Tribes (CSKT) was established to engage the community as partners and advisors in pharmacogenetic research. METHODS: A community advisory committee, the Community Pharmacogenetics Advisory Council (CPAC), was established to ensure community involvement in the research process. To promote bidirectional learning, researchers gave workshops and presentations about pharmacogenetic research to increase research capacity and CPAC members trained researchers in cultural competencies. As part of our commitment to a sustainable relationship, we conducted a self-assessment of the partnership, which included surveys and interviews with CPAC members and researchers. RESULTS: Academic and community participants agree that the partnership has promoted a bidirectional exchange of knowledge. Interviews showed positive feedback from the perspectives of both the CPAC and researchers. CPAC members discussed their trust in and support of the partnership, as well as having learned more about research processes and pharmacogenetics. Researchers discussed their appreciation of CPAC involvement in the project and guidance the group provided in understanding the CSKT community and culture. DISCUSSION: We have created an academic-community partnership to ensure CSKT community input and to share decision making about pharmacogenetic research. Our CBPR approach may be a model for engaging AI/AN people, and other underserved populations, in genetic research.

Pharmacogenomics in diverse practice settings: implementation beyond major metropolitan areas.

AIM: The limited formal study of the clinical feasibility of implementing pharmacogenomic tests has thus far focused on providers at large medical centers in urban areas. Our research focuses on small metropolitan, rural and tribal practice settings. MATERIALS & METHODS: We interviewed 17 healthcare providers in western Montana regarding pharmacogenomic testing. RESULTS: Participants were optimistic about the potential of pharmacogenomic tests, but noted unique barriers in small and rural settings including cost, adherence, patient acceptability and testing timeframe. Participants in tribal settings identified heightened sensitivity to genetics and need for community leadership approval as additional considerations. CONCLUSION: Implementation differences in small metropolitan, rural and tribal communities may affect pharmacogenomic test adoption and utilization, potentially impacting many patients. Original submitted 3 September 2014; Revision submitted 3 December 2014.

Pharmacogenetics in American Indian populations: analysis of CYP2D6, CYP3A4, CYP3A5, and CYP2C9 in the Confederated Salish and Kootenai Tribes.

OBJECTIVES: Cytochrome P450 enzymes play a dominant role in drug elimination and variation in these genes is a major source of interindividual differences in drug response. Little is known, however, about pharmacogenetic variation in American Indian and Alaska Native (AI/AN) populations. We have developed a partnership with the Confederated Salish and Kootenai Tribes (CSKT) in northwestern Montana to address this knowledge gap. METHODS: We resequenced CYP2D6 in 187 CSKT individuals and CYP3A4, CYP3A5, and CYP2C9 in 94 CSKT individuals. RESULTS: We identified 67 variants in CYP2D6, 15 in CYP3A4, 10 in CYP3A5, and 41 in CYP2C9. The most common CYP2D6 alleles were CYP2D6*4 and *41 (20.86 and 11.23%, respectively). CYP2D6*3, *5, *6, *9, *10, *17, *28, *33, *35, *49, *1xN, *2xN, and *4xN frequencies were less than 2%. CYP3A5*3, CYP3A4*1G, and *1B were detected with frequencies of 92.47, 26.81, and 2.20%, respectively. Allelic variation in CYP2C9 was low: CYP2C9*2 (5.17%) and *3 (2.69%). In general, allele frequencies in CYP2D6, CYP2C9, and CYP3A5 were similar to those observed in European Americans. There was, however, a marked divergence in CYP3A4 for the CYP3A4*1G allele. We also observed low levels of linkage between CYP3A4*1G and CYP3A5*1 in the CSKT. The combination of nonfunctional CYP3A5*3 and putative reduced function CYP3A4*1G alleles may predict diminished clearance of CYP3A substrates. CONCLUSION: These results highlight the importance of carrying out pharmacogenomic research in AI/AN populations and show that extrapolation from other populations is not appropriate. This information could help optimize drug therapy for the CSKT population.