ABSTRACT
Despite their crucial role in health and disease, our knowledge of immune cells within human tissues remains limited. We surveyed the immune compartment of 16 tissues from 12 adult donors by single-cell RNA sequencing and VDJ sequencing generating a dataset of ~360,000 cells. To systematically resolve immune cell heterogeneity across tissues, we developed CellTypist, a machine learning tool for rapid and precise cell type annotation. Using this approach, combined with detailed curation, we determined the tissue distribution of finely phenotyped immune cell types, revealing hitherto unappreciated tissue-specific features and clonal architecture of T and B cells. Our multitissue approach lays the foundation for identifying highly resolved immune cell types by leveraging a common reference dataset, tissue-integrated expression analysis, and antigen receptor sequencing.
Subject(s)
B-Lymphocytes , Machine Learning , Sequence Analysis, RNA , Single-Cell Analysis , T-Lymphocytes , Transcriptome , Cells, Cultured , Humans , Organ SpecificityABSTRACT
Statins have been proposed to be a potential treatment for pulmonary arterial hypertension. If introduced into clinical practice, the statin would have to be used in conjunction with established therapy. We investigated the effects of combining simvastatin with a phosphodiesterase type-5 inhibitor, sildenafil, in the rat model of hypoxia-induced pulmonary hypertension. Rats were allocated to either: 1) a prevention protocol, to receive simvastatin 20 mg x kg(-1) x day(-1) by intraperitoneal injection or sildenafil 75 mg x kg(-1) x day(-1) orally or the combination (or vehicle) for 2 weeks beginning at the start of exposure to hypoxia (10% inspired oxygen); or 2) a treatment protocol, where the same agents were administered in the last 2 weeks of a 4-week period of hypoxia. In both protocols, the combination of sildenafil and simvastatin lowered pulmonary artery pressure and produced a significantly greater reduction in right ventricular hypertrophy and pulmonary vascular muscularisation than either drug alone. Moreover, the combination augmented significantly endothelial nitric oxide synthase expression and cGMP levels in the lung and right ventricle above that produced by either drug independently and resulted in greater inhibition of RhoA activity. These data suggest that simvastatin can be usefully combined with sildenafil in the treatment of pulmonary arterial hypertension to achieve greater therapeutic benefit.
Subject(s)
Hydroxymethylglutaryl-CoA Reductase Inhibitors/pharmacology , Hypertension, Pulmonary/drug therapy , Phosphodiesterase Inhibitors/pharmacology , Piperazines/pharmacology , Simvastatin/pharmacology , Sulfones/pharmacology , Animals , Cyclic GMP/metabolism , Disease Models, Animal , Drug Therapy, Combination , Hypertension, Pulmonary/etiology , Hypertrophy, Right Ventricular/drug therapy , Hypertrophy, Right Ventricular/etiology , Hypertrophy, Right Ventricular/prevention & control , Hypoxia/complications , Male , Nitric Oxide Synthase Type III/metabolism , Pulmonary Circulation/drug effects , Purines/pharmacology , Rats , Rats, Sprague-Dawley , Signal Transduction/drug effects , Sildenafil Citrate , rhoA GTP-Binding Protein/metabolismABSTRACT
Selective systems for screening Arabidopsis thaliana (L.) Heynh. mutants with altered sensitivity to the oxidative stress (OS) inductors norflurazon (NF), acifluorfen (AF), and plumbagin (PB) were developed and a collection of 28 mutants was obtained. Dwarf and necrotic forms predominated among the NF-tolerant mutants, while pigment mutants and those with changed root morphology prevailed among the AF-tolerant and PB-sensitive mutants, respectively. Genetic and biochemical analysis of certain mutants was performed; quantitative and qualitative changes in the content of superoxide dismutase and peroxidase isoforms have been revealed. These data, complemented by the data on the cross-tolerance (sensitivity) of the mutants to paraquat, indicate a correlation between tolerance to the OS inductors and the functions of antioxidant systems.
Subject(s)
Arabidopsis/drug effects , Arabidopsis/genetics , Herbicides/pharmacology , Mutation , Oxidative Stress , Arabidopsis/enzymology , Drug Resistance/genetics , Isoenzymes/analysis , Naphthoquinones/pharmacology , Nitrobenzoates/pharmacology , Oxidants/pharmacology , Oxidative Stress/genetics , Peroxidase/analysis , Plant Leaves/drug effects , Plant Leaves/enzymology , Plant Leaves/genetics , Plant Roots/drug effects , Plant Roots/enzymology , Plant Roots/genetics , Pyridazines/pharmacology , Superoxide Dismutase/analysisABSTRACT
The activities of superoxide dismutase and guaiacol-dependent peroxidase were studied in the ontogenesis of recessive homozygous mutants of Arabidopsis thaliana Heynh. le-2 and nfz24, which are characterized by two- to threefold increases in tolerance to the herbicide norflurazone. The mutants le-2 and nfz24 differed from the initial race Dijon in some phenotypic features, duration of ontogenetic stages, and dynamics of the superoxide dismutase and peroxidase activities in ontogenesis. A single treatment of plants with norflurazone induced an accelerated increase in the level of both enzymes in the mutants as compared to the wild type plants. Under the conditions of multiple treatment with norflurazone, the mutants le-2 and nfz24 displayed a higher tolerance to the bleaching effect of the herbicide and were characterized by a higher level of superoxide dismutase. The data obtained suggest that the superoxide dismutase and peroxidase activities are controlled by both ontogenetic factors and stress signals. Mutations in the lines le-2 and nfz24 increase sensitivity to a stress signal or increase efficiency of an adaptive response due to long-term maintenance of a high level of the antioxidant enzymes under the conditions of stress.
