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Chrysanthemum is a very popular flower in Japan and is known to be infected by many soil-borne plant pathogens including nematodes. A nematode survey in six chrysanthemum fields in Okinawa, Japan, found Pratylenchus, Meloidogyne, and Paratylenchus (P. dianthus). The first two genera are known as plant pathogens against chrysanthemum, however, Paratylenchus has not been reported previously. Chrysanthemum seedlings were grown in pots containing soil infected only with P. dianthus for two months in 2017 and 2018. The nematicide imicyafos was applied in triplicates to half of the pots (treated) while the other half were left without the nematicide (non-treated). Plant height and dry plant weight of the imicyafos treated plants exceeded those of the control plants. Also, single-photon avalanche diode value of chrysanthemum leaves was higher in imicyafos treated plants than in the non-treated plants at two-month after planting. The results suggest that P. dianthus may suppress the growth of chrysanthemum. For high-throughput nematode diagnosis, a real-time PCR primer set specific to P. dianthus was developed and its sensitivity to quantify P. dianthus was confirmed with a proper calibration curve. The calibration curve was developed in a simpli- fied approach by using serially diluted DNA extracted from individual nematodes. [ABSTRACT FROM AUTHOR]

• A sensitive assay has been developed to measure human plasma free pazopanib levels using UHPLC-MS/MS. • The assay fulfills the requirements of US Food and Drug Administration for bioanalytical method validation. • The novel method has clinical potential to quantify free pazopanib concentration in patients receiving pazopanib. Pazopanib is widely used to treat renal cell carcinomas and soft tissue tumors in Japan. Although several reports demonstrated the usefulness of therapeutic drug monitoring (TDM) of pazopanib, those studies measured only total pazopanib concentration. For drugs with high protein binding rates such as pazopanib, measuring free concentrations may be clinically more useful than measuring total concentrations. In this study, we aimed to develop a high-throughput method for quantification of free pazopanib in human plasma using ultra-high performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS). Free pazopanib was separated by ultrafiltration. After a simple solid-phase extraction step using a 96-well plate, pazopanib was analyzed by UHPLC-MS/MS in positive electrospray ionization mode. The novel method fulfilled the requirements of the US Food and Drug Administration guidelines for assay validation, and the lower limit of quantification was 0.05 ng/mL. The calibration curve was linear over the concentration range of 0.05–50 ng/mL. The average recovery rate was 66.9 ± 2.1% (mean ± SD). The precision was below 7.02%, and accuracy was within 10.60% across all quality control levels. Matrix effect varied between 44.4% and 60.4%. This assay was successfully applied to measure trough free pazopanib concentrations in three patients treated with pazopanib for soft tissue tumors. We succeeded to develop a novel high-throughput UHPLC-MS/MS method for quantification of free pazopanib in human plasma. This method can be applied to TDM for patients receiving pazopanib in the clinical setting. [ABSTRACT FROM AUTHOR]