Evaluating imputation methods for single-cell RNA-seq data.

BACKGROUND: Single-cell RNA sequencing (scRNA-seq) enables the high-throughput profiling of gene expression at the single-cell level. However, overwhelming dropouts within data may obscure meaningful biological signals. Various imputation methods have recently been developed to address this problem. Therefore, it is important to perform a systematic evaluation of different imputation algorithms. RESULTS: In this study, we evaluated 11 of the most recent imputation methods on 12 real biological datasets from immunological studies and 4 simulated datasets. The performance of these methods was compared, based on numerical recovery, cell clustering and marker gene analysis. Most of the methods brought some benefits on numerical recovery. To some extent, the performance of imputation methods varied among protocols. In the cell clustering analysis, no method performed consistently well across all datasets. Some methods performed poorly on real datasets but excellent on simulated datasets. Surprisingly and importantly, some methods had a negative effect on cell clustering. In marker gene analysis, some methods identified potentially novel cell subsets. However, not all of the marker genes were successfully imputed in gene expression, suggesting that imputation challenges remain. CONCLUSIONS: In summary, different imputation methods showed different effects on different datasets, suggesting that imputation may have dataset specificity. Our study reveals the benefits and limitations of various imputation methods and provides a data-driven guidance for scRNA-seq data analysis.

Combined acid rain and lanthanum pollution and its potential ecological risk for nitrogen assimilation in soybean seedling roots.

Rare earth elements (REEs) are used in various fields, resulting in their accumulation in the environment. This accumulation has affected the survival and distribution of crops in various ways. Acid rain is a serious global environmental problem. The combined effects on crops from these two types of pollution have been reported, but the effects on crop root nitrogen assimilation are rarely known. To explore the impact of combined contamination from these two pollutants on crop nitrogen assimilation, the soybean seedlings were treated with simulated environmental pollution from acid rain and a representative rare earth ion, lanthanum ion (La3+), then the indexes related to plant nitrogen assimilation process in roots were determined. The results showed that combined treatment with pH 4.5 acid rain and 0.08 mM La3+ promoted nitrogen assimilation synergistically, while the other combined treatments all showed inhibitory effects. Moreover, acid rain aggravated the inhibitory effect of 1.20 or 0.40 mM La3+ on nitrogen assimilation in soybean seedling roots. Thus, the effects of acid rain and La3+ on crops depended on the combination levels of acid rain intensity and La3+ concentration. Acid rain increases the bioavailability of La3+, and the combined effects of these two pollutants were more serious than that of either pollutant alone. These results provide new evidence in favor of limiting overuse of REEs in agriculture. This work also provides a new framework for ecological risk assessment of combined acid rain and REEs pollution on soybean crops.

Analysis of the combined effects of lanthanum and acid rain, and their mechanisms, on nitrate reductase transcription in plants.

Rare earth element (REE) pollution and acid rain are major global environmental concerns, and their spatial distributions overlap. Thus, both forms of pollution combine to act on plants. Nitrogen is important for plant growth, and nitrate reductase (NR) is a key plant enzyme that catalyzes nitrogen assimilation. Studying the combined effects of REEs and acid rain on plant nitrogen-based nutrients has important environmental significance. Here, soybean (Glycine max) plants, commonly used for toxicological studies, were exposed to lanthanum (La), a REE, and acid rain to study the NR activities and NR transcriptional levels in the roots. To explain how the pollution affected the NR transcriptional level, we simultaneously observed the contents of intracellular La and nutrient elements, protoplast morphology, membrane lipid peroxidation and intracellular pH. A combined treatment of 0.08mmol/L La and pH 4.5 acid rain increased the NR activity, decreased the NR transcriptional level, increased the intracellular nutrient elements' contents and caused deformations in membrane structures. Other combined treatments significantly decreased the aforementioned parameters and caused serious damage to the membrane structures. The variation in the amplitudes of combined treatments was greater than those of individual treatments. Compared with the control and individual treatments, combined treatments increased membrane permeability, the malondialdehyde content, and intracellular H+ and La contents, and with an increasing La concentration or acid strength, the change in amplitude increased. Thus, the combined effects on NR gene transcription in soybean seedling roots were related to the intracellular nutrient elements' contents, protoplast morphology, membranous lipid peroxidation, intracellular pH and La content.

Combined Effects of Lanthanum (III) and Acid Rain on Antioxidant Enzyme System in Soybean Roots.

Rare earth element pollution (REEs) and acid rain (AR) pollution simultaneously occur in many regions, which resulted in a new environmental issue, the combined pollution of REEs and AR. The effects of the combined pollution on the antioxidant enzyme system of plant roots have not been reported. Here, the combined effects of lanthanum ion (La3+), one type of REE, and AR on the antioxidant enzyme system of soybean roots were investigated. In the combined treatment of La3+ (0.08 mM) and AR, the cell membrane permeability and the peroxidation of cell membrane lipid of soybean roots increased, and the superoxide dismutase, catalase, peroxidase and reduced ascorbic acid served as scavengers of reactive oxygen species. In other combined treatments of La3+ (0.40 mM, 1.20 mM) and AR, the membrane permeability, malonyldialdehyde content, superoxide dismutase activity, peroxidase activity and reduced ascorbic acid content increased, while the catalase activity decreased. The increased superoxide dismutase activity, peroxidase activity and reduced ascorbic acid content were inadequate to scavenge the excess hydrogen peroxide and superoxide, leading to the damage of the cell membrane, which was aggravated with the increase in the concentration of La3+ and the level of AR. The deleterious effects of the combined treatment of La3+ and AR were stronger than those of the single treatment of La3+ or AR. Moreover, the activity of antioxidant enzyme system in the combined treatment group was affected directly and indirectly by mineral element content in soybean plants.

Combined effects of Lanthanum(III) and elevated Ultraviolet-B radiation on root nitrogen nutrient in soybean seedlings.

Rare earth element pollution and elevated ultraviolet-B (UV-B) radiation occur simultaneously in some regions, but the combined effects of these two factors on plants have not attracted enough attention. Nitrogen nutrient is vital to plant growth. In this study, the combined effects of lanthanum(III) and elevated UV-B radiation on nitrate reduction and ammonia assimilation in soybean (Glycine max L.) roots were investigated. Treatment with 0.08 mmol L(-1) La(III) did not change the effects of elevated UV-B radiation on nitrate reductase (NR), nitrite reductase (NiR), glutamine synthetase (GS), glutamate synthase (GOGAT), glutamate dehydrogenase (GDH), nitrate, ammonium, amino acids, or soluble protein in the roots. Treatment with 0.24 mmol L(-1) La(III) and elevated UV-B radiation synergistically decreased the NR, NiR, GS, and GOGAT activities as well as the nitrate, amino acid, and soluble protein levels, except for the GDH activity and ammonium content. Combined treatment with 1.20 mmol L(-1) La(III) and elevated UV-B radiation produced severely deleterious effects on all test indices, and these effects were stronger than those induced by La(III) or elevated UV-B radiation treatment alone. Following the withdrawal of La(III) and elevated UV-B radiation, all test indices for the combined treatments with 0.08/0.24 mmol L(-1) La(III) and elevated UV-B radiation recovered to a certain extent, but they could not recover for treatments with 1.20 mmol L(-1) La(III) and elevated UV-B radiation. In summary, combined treatment with La(III) and elevated UV-B radiation seriously affected nitrogen nutrition in soybean roots through the inhibition of nitrate reduction and ammonia assimilation.

Combined effects of bisphenol A and cadmium on growth and nitrate assimilation of soybean seedling roots.

Bisphenol A (BPA) and cadmium (Cd) pollution exist simultaneously in many regions. However, little information is available regarding the combined effects of BPA and Cd pollution on plants. Plant roots are in direct contact with the soil, which is an important compartment of BPA and Cd. In the present study, the effects of combined BPA and Cd pollution on soybean seedling roots were evaluated in pot experiments. Combined treatment with BPA and Cd at low concentrations (1.5 mg/kg BPA and 0.2 mg/kg Cd) improved soybean seedling root growth. However, other combined BPA and Cd treatments, including combined treatment with BPA (Cd) at the low concentration and Cd (BPA) at the high concentration as well as combined treatment with BPA and Cd at the high concentration, inhibited soybean seedling root growth. The improvement or inhibition of soybean seedling root growth was greater in the combined BPA and Cd treatments than in single treatments. The effects of the combined BPA and Cd treatments on root growth resulted from changes in nitrate assimilation. In addition, the combined effects of BPA and Cd on the nitrate and ammonium contents in roots are discussed. The present research provides a basic understanding of the combined effects of BPA and Cd pollution on plant roots.

Effects and mechanisms of the combined pollution of lanthanum and acid rain on the root phenotype of soybean seedlings.

Rare earth pollution and acid rain pollution are both important environmental issues worldwide. In regions which simultaneously occur, the combined pollution of rare earth and acid rain becomes a new environmental issue, and the relevant research is rarely reported. Accordingly, we investigated the combined effects and mechanisms of lanthanum ion (La(3+)) and acid rain on the root phenotype of soybean seedlings. The combined pollution of low-concentration La(3+) and acid rain exerted deleterious effects on the phenotype and growth of roots, which were aggravated by the combined pollution of high-concentration La(3+) and acid rain. The deleterious effects of the combined pollution were stronger than those of single La(3+) or acid rain pollution. These stronger deleterious effects on the root phenotype and growth of roots were due to the increased disturbance of absorption and utilization of mineral nutrients in roots.

Living target of Ce(III) action on horseradish cells: proteins on/in cell membrane.

Positive and negative effects of rare earth elements (REEs) in life have been reported in many papers, but the cellular mechanisms have not been answered, especially the action sites of REEs on plasma membrane are unknown. Proteins on/in the plasma membrane perform main functions of the plasma membrane. Cerium (Ce) is the richest REEs in crust. Thus, the interaction between Ce(III) and the proteins on/in the plasma membrane, the morphology of protoplast, and the contents of nutrient elements in protoplast of horseradish were investigated using the optimized combination of the fluorescence microscopy, fluorescence spectroscopy, circular dichroism, scanning electron microscopy, and X-ray energy dispersive spectroscopy. It was found that Ce(III) at the low concentrations (10, 30 µM) could interact with proteins on/in the plasma membrane of horseradish, leading to the improvement in the structure of membrane proteins and the plasma membrane, which accelerated the intra-/extra-cellular substance exchange and further promoted the development of cells. When horseradish was treated with Ce(III) at the high concentrations (60, 80 µM), Ce(III) also could interact with the proteins on/in the plasma membrane of horseradish, leading to the destruction in the structure of membrane proteins and the plasma membrane. These effects decelerated the intra-/extra-cellular substance exchange and further inhibited the development of cells. Thus, the interaction between Ce(III) and proteins on/in the plasma membrane in plants was an important reason of the positive and negative effects of Ce(III) on plants. The results would provide some references for understanding the cellular effect mechanisms of REEs on plants.

Interactive effects of cadmium and acid rain on photosynthetic light reaction in soybean seedlings.

Interactive effects of cadmium (Cd(2+)) and acid rain on photosynthetic light reaction in soybean seedlings were investigated under hydroponic conditions. Single treatment with Cd(2+) or acid rain and the combined treatment decreased the content of chlorophyll, Hill reaction rate, the activity of Mg(2+)-ATPase, maximal photochemical efficiency and maximal quantum yield, increased initial fluorescence and damaged the chloroplast structure in soybean seedlings. In the combined treatment, the change in the photosynthetic parameters and the damage of chloroplast structure were stronger than those of any single pollution. Meanwhile, Cd(2+) and acid rain had the interactive effects on the test indices in soybean seedlings. The results indicated that the combined pollution of Cd(2+) and acid rain aggravated the toxic effect of the single pollution of Cd(2+) or acid rain on the photosynthetic parameters due to the serious damage to the chloroplast structure.