Specialized pro-resolvin mediators induce cell growth and improve wound repair in intestinal epithelial Caco-2 cell cultures.

Specialized pro-resolvin mediators (SPMs) are a superfamily of bioactive molecules synthesized from polyunsaturated fatty acids (arachidonic, eicosapentaenoic and docosahexaenoic acids) that include resolvins, protectins and maresins. These metabolites are important to control the resolution phase of inflammation and the epithelial repair, which is essential in restoring the mucosal barriers. Unfortunately, the effects of SPMs on intestinal epithelial cell growth remain poorly understood. Caco-2 cell were used as intestinal epithelial cell model. Cell growth/DNA synthesis, cell signalling pathways, western blot and wound repair assay were performed. Our data demonstrated that SPMs such as lipoxin LxA4, resolvin (Rv) E1, RvD1, protectin D 1 and maresin 1 were able to enhance intestinal epithelial Caco-2 cell growth and DNA synthesis. Furthermore, our results provide evidence that these effects of RvE1 and RvD1 were associated with a pertussis toxin-sensitive G protein-coupled receptor, and that leukotriene B4 receptor 2 could be involved, at least in part, in these effects of RvE1/RvD1. Moreover, these mitogenic effects induced by SPMs were dependent on the ERK 1/2 and p38 MAPK pathways as well as phospholipase C and protein kinase C activation. Thus, these mitogenic effects of RvE1/RvD1 on intestinal epithelial cells could be involved in this signalling circuit involved in wounded epithelium and the catabasis process. Keywords: Lipoxin; Maresin; Protectin; Resolvin; Wound closure

Pii‐21

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/109873/1/cptclpt2006159.pd

Pi‐29

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/110026/1/cptclpt200664.pd

Pii‐18

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/110052/1/cptclpt2006156.pd

Whole Genome Amplification of DNA for Genotyping Pharmacogenetics Candidate Genes

Whole genome amplification (WGA) technologies can be used to amplify genomic DNA when only small amounts of DNA are available. The Multiple Displacement Amplification Phi polymerase based amplification has been shown to accurately amplify DNA for a variety of genotyping assays; however, it has not been tested for genotyping many of the clinically relevant genes important for pharmacogenetic studies, such as the cytochrome P450 genes, that are typically difficult to genotype due to multiple pseudogenes, copy number variations, and high similarity to other related genes. We evaluated whole genome amplified samples for Taqman™ genotyping of SNPs in a variety of pharmacogenetic genes. In 24 DNA samples from the Coriell human diversity panel, the call rates, and concordance between amplified (∼200-fold amplification) and unamplified samples was 100% for two SNPs in CYP2D6 and one in ESR1. In samples from a breast cancer clinical trial (Trial 1), we compared the genotyping results in samples before and after WGA for three SNPs in CYP2D6, one SNP in CYP2C19, one SNP in CYP19A1, two SNPs in ESR1, and two SNPs in ESR2. The concordance rates were all >97%. Finally, we compared the allele frequencies of 143 SNPs determined in Trial 1 (whole genome amplified DNA) to the allele frequencies determined in unamplified DNA samples from a separate trial (Trial 2) that enrolled a similar population. The call rates and allele frequencies between the two trials were 98 and 99.7%, respectively. We conclude that the whole genome amplified DNA is suitable for Taqman™ genotyping for a wide variety of pharmacogenetically relevant SNPs

Evaluación de las características hidroquímicas de antiguos cauces del Río Dulce en Villa Nueva, provincia de Santiago del Estero

El acceso al agua segura, en pequeñas localidades como Villa Nueva, es un problema de vieja data. El Área de estudio está situada en el sector SO de la provincia de Santiago del Estero, y geomorfológicamente se ubica en el tramo medio de la Llanura Aluvial del Río Dulce. Se caracteriza por una sucesión de antiguos cauces colmatados y entrelazados. El trabajo de investigación se orientó hacia la caracterización hidroquímica del acuífero libre, y su utilización como soporte de almacenamiento y de recarga en forma natural o artificial, para el abastecimiento de agua destinada al consumo humano. Mediante perforaciones se realizaron estudios granulométricos de los sedimentos que permitieron obtener una aproximación acerca de la permeabilidad. Los análisis físico-químicos de las muestras de agua obtenidas, no superan los límites tolerables de las aguas seguras, excepto una de ellas, que supera la concentración de sulfatos.Access to safe water, in small towns like Villa Nueva, is a long-standing problem. The research area is located in the SW sector of the province of Santiago del Estero, and geomorphologically is located in the middle stretch of the Dulce River Floodplain. It is characterized by a succession of former channels silted and intertwined. The research work was oriented towards free aquifer hydrochemical characterization, and their use as storage media and recharge in natural or artificial, for the supply of water for human consumption. Through perforations were made sediments grain size determination studies, we have obtained an approximation concerning permeability. The physico-chemical analysis of water samples obtained do not exceed tolerable limits of safe water, except one, which exceeds the concentration of sulfates.Universidad Nacional de La Plat

Plasma Letrozole Concentrations in Postmenopausal Women With Breast Cancer Are Associated With CYP2A6 Genetic Variants, Body Mass Index, and Age

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/109858/1/cptclpt2011174.pd

Effects of exemestane and letrozole therapy on plasma concentrations of estrogens in a randomized trial of postmenopausal women with breast cancer

PURPOSE: Inter-individual differences in estrogen concentrations during treatment with aromatase inhibitors (AIs) may contribute to therapeutic response and toxicity. The aim of this study was to determine plasma concentrations of estradiol (E2), estrone (E1), and estrone sulfate (E1S) in a large cohort of AI-treated breast cancer patients. METHODS: In a randomized, multicenter trial of postmenopausal women with early-stage breast cancer starting treatment with letrozole (n = 241) or exemestane (n = 228), plasma estrogen concentrations at baseline and after 3 months were quantitated using a sensitive mass spectrometry-based assay. Concentrations and suppression below the lower limit of quantification (LLOQ) were compared between estrogens and between drugs. RESULTS: The ranges of baseline estrogen concentrations were <LLOQ-361 pg/mL for E2, <LLOQ-190 pg/mL for E1, and 8.3-4060 pg/mL for E1S. For E2, the frequency of suppression below the LLOQ was not statistically significantly different between AIs (exemestane: 89.0%, letrozole: 86.9%, p = 0.51). However, patients on letrozole were more likely to achieve suppression below the LLOQ of both E1 (exemestane: 80.1%, letrozole: 90.1%, p = 0.005) and E1S (exemestane: 17.4%, letrozole: 54.9%, p = 4.34e-15). After 3 months of AI therapy, the ranges of estrogen concentrations were <LLOQ-63.8 pg/mL, <LLOQ-36.7 pg/mL, and <LLOQ-1090 pg/mL for E2, E1, and E1S, respectively. During treatment, 16 patients had an increased concentration compared to the baseline concentration of at least one estrogen. CONCLUSIONS: Letrozole had greater suppression of plasma E1 and E1S than exemestane, though the response was highly variable among patients. Additional research is required to examine the clinical relevance of differential estrogen suppression

Prospective assessment of patient-reported outcomes and estradiol and drug concentrations in patients experiencing toxicity from adjuvant aromatase inhibitors

PURPOSE: Aromatase inhibitors (AI), which decrease circulating estradiol concentrations in post-menopausal women, are associated with toxicities that limit adherence. Approximately one-third of patients will tolerate a different AI after not tolerating the first. We report the effect of crossover from exemestane to letrozole or vice versa on patient-reported outcomes (PROs) and whether the success of crossover is due to lack of estrogen suppression. METHODS: Post-menopausal women enrolled on a prospective trial initiating AI therapy for early-stage breast cancer were randomized to exemestane or letrozole. Those that discontinued for intolerance were offered protocol-directed crossover to the other AI after a washout period. Changes in PROs, including pain [Visual Analog Scale (VAS)] and functional status [Health Assessment Questionnaire (HAQ)], were compared after 3 months on the first versus the second AI. Estradiol and drug concentrations were measured. RESULTS: Eighty-three patients participated in the crossover protocol, of whom 91.3% reported improvement in symptoms prior to starting the second AI. Functional status worsened less after 3 months with the second AI (HAQ mean change AI #1: 0.2 [SD 0.41] vs. AI #2: -0.05 [SD 0.36]; p = 0.001); change in pain scores was similar between the first and second AI (VAS mean change AI #1: 0.8 [SD 2.7] vs. AI #2: -0.2 [SD 2.8]; p = 0.19). No statistical differences in estradiol or drug concentrations were found between those that continued or discontinued AI after crossover. CONCLUSIONS: Although all AIs act via the same mechanism, a subset of patients intolerant to one AI report improved PROs with a different one. The mechanism of this tolerance remains unknown, but does not appear to be due to non-adherence to, or insufficient estrogen suppression by, the second AI