Untargeted mass spectrometry (MS) experiments produce complex, multidimensional data that are practically impossible to investigate manually. For this reason, computational pipelines are needed to extract relevant information from raw spectral data and convert it into a more comprehensible format. Depending on the sample type and/or goal of the study, a variety of MS platforms can be used for such analysis. MZmine is an open-source software for the processing of raw spectral data generated by different MS platforms. Examples include liquid chromatography-MS, gas chromatography-MS and MS-imaging. These data might typically be associated with various applications including metabolomics and lipidomics. Moreover, the third version of the software, described herein, supports the processing of ion mobility spectrometry (IMS) data. The present protocol provides three distinct procedures to perform feature detection and annotation of untargeted MS data produced by different instrumental setups: liquid chromatography-(IMS-)MS, gas chromatography-MS and (IMS-)MS imaging. For training purposes, example datasets are provided together with configuration batch files (i.e., list of processing steps and parameters) to allow new users to easily replicate the described workflows. Depending on the number of data files and available computing resources, we anticipate this to take between 2 and 24 h for new MZmine users and nonexperts. Within each procedure, we provide a detailed description for all processing parameters together with instructions/recommendations for their optimization. The main generated outputs are represented by aligned feature tables and fragmentation spectra lists that can be used by other third-party tools for further downstream analysis.

• MeSH
• chromatografie kapalinová metody   MeSH
• hmotnostní spektrometrie * metody   MeSH
• iontová mobilní spektrometrie metody   MeSH
• metabolomika metody   MeSH
• plynová chromatografie s hmotnostně spektrometrickou detekcí metody   MeSH
• reprodukovatelnost výsledků MeSH
• software * MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH

• MeSH
• hmotnostní spektrometrie metody   MeSH
• software * MeSH
• Publikační typ
• dopisy MeSH

Insect herbivores have evolved a broad spectrum of adaptations in response to the diversity of chemical defences employed by plants. Here we focus on two species of New Guinean Asota and determine how these specialist moths deal with the leaf alkaloids of their fig (Ficus) hosts. As each focal Asota species is restricted to one of three chemically distinct species of Ficus, we also test whether these specialized interactions lead to similar alkaloid profiles in both Asota species. We reared Asota caterpillars on their respective Ficus hosts in natural conditions and analyzed the alkaloid profiles of leaf, frass, caterpillar, and adult moth samples using UHPLC-MS/MS analyses. We identified 43 alkaloids in our samples. Leaf alkaloids showed various fates. Some were excreted in frass or found in caterpillars and adult moths. We also found two apparently novel indole alkaloids-likely synthesized de novo by the moths or their microbiota-in both caterpillar and adult tissue but not in leaves or frass. Overall, alkaloids unique or largely restricted to insect tissue were shared across moth species despite feeding on different hosts. This indicates that a limited number of plant compounds have a direct ecological function that is conserved among the studied species. Our results provide evidence for the importance of phytochemistry and metabolic strategies in the formation of plant-insect interactions and food webs in general. Furthermore, we provide a new potential example of insects acquiring chemicals for their benefit in an ecologically relevant insect genus.

• Klíčová slova
• Alkaloids, Biodiversity, Food-webs, Herbivores, Host-specificity, Plant–insect interactions,
• MeSH
• alkaloidy * MeSH
• býložravci MeSH
• Ficus * MeSH
• hmyz MeSH
• larva fyziologie   MeSH
• metabolom MeSH
• můry * fyziologie   MeSH
• rostliny MeSH
• tandemová hmotnostní spektrometrie MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Nová Guinea MeSH
• Názvy látek
• alkaloidy * MeSH

Cytochrome P450 (P450) 2U1 has been shown to be expressed, at the mRNA level, in human thymus, brain, and several other tissues. Recombinant P450 2U1 was purified and used as a reagent in a metabolomic search for substrates in bovine brain. In addition to fatty acid oxidation reactions, an oxidation of endogenous N-arachidonoylserotonin was characterized. Subsequent NMR and mass spectrometry and chemical synthesis showed that the main product was the result of C-2 oxidation of the indole ring, in contrast to other human P450s that generated different products. N-Arachidonoylserotonin, first synthesized chemically and described as an inhibitor of fatty acid amide hydrolase, had previously been found in porcine and mouse intestine; we demonstrated its presence in bovine and human brain samples. The product (2-oxo) was 4-fold less active than N-arachidonoylserotonin in inhibiting fatty acid amide hydrolase. The rate of oxidation of N-arachidonoylserotonin was similar to that of arachidonic acid, one of the previously identified fatty acid substrates of P450 2U1. The demonstration of the oxidation of N-arachidonoylserotonin by P450 2U1 suggests a possible role in human brain and possibly other sites.

• Klíčová slova
• Arachidonic Acid, Cytochrome P450, Eicosanoid, Lipid Oxidation, Mass Spectrometry (MS),
• MeSH
• chromatografie kapalinová MeSH
• erytrocyty enzymologie   MeSH
• Escherichia coli metabolismus   MeSH
• hmotnostní spektrometrie MeSH
• indoly metabolismus   MeSH
• játra enzymologie   MeSH
• katalýza MeSH
• kyseliny arachidonové metabolismus   MeSH
• kyslík metabolismus   MeSH
• lidé MeSH
• magnetická rezonanční spektroskopie MeSH
• molekulární sekvence - údaje MeSH
• mozek metabolismus   MeSH
• polymerázová řetězová reakce MeSH
• proteomika MeSH
• regulace genové exprese enzymů MeSH
• rodina 2 cytochromů P450 MeSH
• sekvence aminokyselin MeSH
• sekvenční homologie aminokyselin MeSH
• serotonin analogy a deriváty   metabolismus   MeSH
• skot MeSH
• systém (enzymů) cytochromů P-450 metabolismus   MeSH
• vazba proteinů MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• skot MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH
• Názvy látek
• arachidonoylserotonin MeSH Prohlížeč
• CYP2U1 protein, human MeSH Prohlížeč
• indoly MeSH
• kyseliny arachidonové MeSH
• kyslík MeSH
• rodina 2 cytochromů P450 MeSH
• serotonin MeSH
• systém (enzymů) cytochromů P-450 MeSH