Separated Transcriptomes of Male Gametophyte and Tapetum in Rice: Validity of a Laser Microdissection (LM) Microarray

In flowering plants, the male gametophyte, the pollen, develops in the anther. Complex patterns of gene expression in both the gametophytic and sporophytic tissues of the anther regulate this process. The gene expression profiles of the microspore/pollen and the sporophytic tapetum are of particular interest. In this study, a microarray technique combined with laser microdissection (44K LM-microarray) was developed and used to characterize separately the transcriptomes of the microspore/pollen and tapetum in rice. Expression profiles of 11 known tapetum specific-genes were consistent with previous reports. Based on their spatial and temporal expression patterns, 140 genes which had been previously defined as anther specific were further classified as male gametophyte specific (71 genes, 51%), tapetum-specific (seven genes, 5%) or expressed in both male gametophyte and tapetum (62 genes, 44%). These results indicate that the 44K LM-microarray is a reliable tool to analyze the gene expression profiles of two important cell types in the anther, the microspore/pollen and tapetum

Characteristics of Pemetrexed Transport by Renal Basolateral Organic Anion Transporter hOAT3

application/pdfPurpose: Pemetrexed transport by human organic anion transporters, hOAT1 (SLC22A6) and hOAT3 (SLC22A8), were characterized in comparison with methotrexate. Methods: Accumulation of pemetrexed and methotrexate in hOAT1- and hOAT3-expressing cells were evaluated. Pemetrexed and methotrexate were determined by HPLC. Kinetic parameters were calculated by Eadie-Hofstee plot. Results: When HEK-hOAT3 and -hOAT1 cells were incubated with 100 μM pemetrexed for 30min, pemetrexed was accumulated at 14- and 1.7-fold greater than that in control cells, respectively. Pemetrexed and methotrexate transport by hOAT3 was saturated at high concentrations with apparent Km values 28.2 μM and 76.6 μM, respectively. In addition, intrinsic activity (Vmax/Km) of pemetrexed and methotrexate transport by hOAT3 was 4.82 and 0.42 μl/min/mg protein, respectively, suggesting 11-fold higher transport of pemetrexed than methotrexate by hOAT3. Furthermore, loxoprofen, ibuprofen, pravastatin, and cefazolin, transport substrates of hOAT3, inhibited pemetrexed transport by hOAT3 with IC50 values, 34.2, 27.9, 76.3 and 650 μM, respectively. Conclusions: Pemetrexed is a superior substrate to methotrexate for hOAT3. Loxoprofen, ibuprofen, and cefazolin could cause drug-drug interactions when attaining high blood concentrations.本文 / Department of Clinical Pharmacy and Biopharmaceutics, Mie University Graduate School of Medicine6

Characteristics of Pemetrexed Transport by Renal Basolateral Organic Anion Transporter hOAT3

application/pdf内容の要旨・審査結果の要旨 / 三重大学大学院医学系研究科　生命医科学専攻　病態解明医学講座　臨床薬剤学分

Design and Synthesis of New Stable Fluorenyl-Based Radicals

Organic neutral radicals have long fascinated chemists with a fundamental understanding of structure–reactivity relationships in organic reactions and with applications as new functional materials. However, the elusive nature of these radicals makes the synthesis, isolation, and characterization very challenging. In this work, the synthesis of three long-lived, fluorenyl-based radicals are reported. The geometry and electronic structures of these radicals were systematically investigated with a combination of various experimental methods, besides density functional theory (DFT) calculations, which include X-ray crystallographic analysis, electron spin resonance (ESR), electron nuclear double resonance (ENDOR), cyclic voltammetry, and UV–vis–NIR measurements. Their half-life periods (τ_1/2) in air-saturated solution under ambient conditions were also determined. Surprisingly, all three radicals showed remarkable stabilities: τ_1/2 = 7, 3.5, and 43 days