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To the Editor: Paper spray (PS)1 is a recently described method for the direct mass spectrometry (MS) analysis of blood and other biological samples (1–3). Paper spray is performed by depositing a sample such as whole blood onto a paper substrate contained within a disposable cartridge; extraction and ionization occur directly from the disposable cartridge with an automated MS attachment (Fig. 1). No punching or offline extraction is required. Total analysis time, including sample extraction, is several minutes. Fig. 1. Paper spray for direct analysis of blood spots by MS. Ion chronograms from the lowest calibrator in whole blood, overlaid with representative chronograms from blank blood, are shown for cyclosporine and sirolimus. Quantification was performed with the entire area under the curve for each analyte, normalized by SIL-IS, to obtain calibration curves as shown. Because of its simplicity, paper spray could lower the barrier for implementation of MS-based assays in clinical laboratories. We previously used PS-MS/MS to monitor tacrolimus in clinical samples and showed good correlation with 2 immunoassays and an external HPLC-MS/MS method (4). Here we extend this approach by demonstrating the simultaneous quantification of cyclosporine and sirolimus by PS-MS/MS. Therapeutic drug monitoring of these drugs is important for organ transplant patient care. The method used was similar to our …

Background: Mutations in LRRK2 are associated with familial and sporadic Parkinson’s disease (PD). Subjects with PD caused by LRRK2 mutations show pleiotropic pathology that can involve inclusions containing α-synuclein, tau or neither protein. The mechanisms by which mutations in LRRK2 lead to this pleiotropic pathology remain unknown. Objectives: To investigate mechanisms by which LRRK2 might cause PD. Methods: We used systems biology to investigate the transcriptomes from human brains, human blood cells and Caenorhabditis elegans expressing wild-type LRRK2. The role of autophagy was tested in lines of C. elegans expressing LRRK2, V337M tau or both proteins. Neuronal function was measured by quantifying thrashing. Results: Genes regulating autophagy were coordinately regulated with LRRK2. C. elegans expressing V337M tau showed reduced thrashing, as has been noted previously. Coexpressing mutant LRRK2 (R1441C or G2019S) with V337M tau increased the motor deficits. Treating the lines of C. elegans with an mTOR inhibitor that enhances autophagic flux, ridaforolimus, increased the thrashing behavior to the same level as nontransgenic nematodes. Conclusion: These data support a role for LRRK2 in autophagy, raise the possibility that deficits in autophagy contribute to the pathophysiology of LRRK2, and point to a potential therapeutic approach addressing the pathophysiology of LRRK2 in PD.