To improve and complete our knowledge of archaeal tRNA modification patterns, we have identified and compared the modification pattern (type and location) in tRNAs of three very different archaeal species,Methanococcus maripaludis(a mesophilic methanogen),Pyrococcus furiosus(a hyperthermophile thermococcale), andSulfolobus acidocaldarius(an acidophilic thermophilic sulfolobale). Most abundant isoacceptor tRNAs (79 in total) for each of the 20 amino acids were isolated by two-dimensional gel electrophoresis followed by in-gel RNase digestions. The resulting oligonucleotide fragments were separated by nanoLC and their nucleotide content analyzed by mass spectrometry (MS/MS). Analysis of total modified nucleosides obtained from complete digestion of bulk tRNAs was also performed. Distinct base- and/or ribose-methylations, cytidine acetylations, and thiolated pyrimidines were identified, some at new positions in tRNAs. Novel, some tentatively identified, modifications were also found. The least diversified modification landscape is observed in the mesophilicMethanococcus maripaludisand the most complex one inSulfolobus acidocaldarius. Notable observations are the frequent occurrence of ac4C nucleotides in thermophilic archaeal tRNAs, the presence of m7G at positions 1 and 10 inPyrococcus furiosustRNAs, and the use of wyosine derivatives at position 37 of tRNAs, especially those decoding U1- and C1-starting codons. These results complete those already obtained by others with sets of archaeal tRNAs fromMethanocaldococcus jannaschiiandHaloferax volcanii.
Background In neurodegenerative diseases, alongside genetic factors, the possible intervention of environmental factors in the pathogenesis is increasingly being considered. In particular, recent evidence suggests the intervention of a pesticide-like xenobiotic in the initiation of disease with Lewy bodies (DLB). Objectives To test for the presence of pesticides or other xenobiotics in the cerebrospinal fluid (CSF) of patients with DLB. Methods A total of 45 patients were included in this study: 16 patients with DLB at the prodromal stage, 8 patients with DLB at the demented stage, 8 patients with Alzheimer’s disease (AD) at the prodromal stage and 13 patients with AD at the demented stage. CSF was obtained by lumbar puncture and analysed by liquid chromatography–mass spectrometry. Results Among the compounds detected in greater abundance in the CSF of patients with DLB compared with patients with AD, only one had a xenobiotic profile potentially related to the pathophysiology of DLB. After normalisation and scaling, bis(2-ethylhexyl) phthalate was more abundant in the CSF of patients with DLB (whole cohort: 2.7-fold abundant in DLB, p=0.031; patients with dementia: 3.8-fold abundant in DLB, p=0.001). Conclusions This study is the first reported presence of a phthalate in the CSF of patients with DLB. This molecule, which is widely distributed in the environment and enters the body orally, nasally and transdermally, was first introduced in the 1920s as a plasticizer. Thereafter, the first cases of DLB were described in the 1960s and 1970s. These observations suggest that phthalates may be involved in the pathophysiology of DLB.
Micropollutants and emerging organic contaminants (EOCs) have been widely studied in terms of persistance, removal, human risk assessment, toxicology, etc. Mass spectrometry imaging (MSI) offers the possibility of following the fate of a single pesticide in a plant leaf or a drug in the whole body of an animal, organ by organ. However, the admissibility of chronic low doses of complex mixtures for the ecosystem has not been assessed. How do micropollutants diffuse in the environment? How do living organisms cope with chronic exposure to a low dose of diverse micropollutants? Is there a cocktail effect or a chance for hormesis? Combining mass spectrometry imaging (MSI) and targeted and nontargeted liquid chromatography coupled to mass spectrometry (LC-MS), we attempt to answer these questions. We investigate the diversity of micropollutants at the exit of a water treatment facility, their diffusion in sludge and black poplar (Populus nigra), and their impact on a living organism. We reveal a specific tissue localization of micropollutants in peripheral leaf tissues, and an associated stress response from the plant, with stress hormones and tissue degradation markers induced in the plant growing near the water efflux.