Long-term exposure to 6-PPD quinone at environmentally relevant concentrations causes neurotoxicity by affecting dopaminergic, serotonergic, glutamatergic, and GABAergic neuronal systems in Caenorhabditis elegans.

6-PPD quinone (6-PPDQ), an emerging environmental pollutant, is converted based on 6-PPD via ozonation. However, a systematic evaluation on possible neurotoxicity of long-term and low-dose 6-PPDQ exposure and the underlying mechanism remain unknown. In the present work, 0.1-10 µg/L 6-PPDQ was added to treat Caenorhabditis elegans for 4.5 days, with locomotion behavior, neuronal development, sensory perception behavior, neurotransmitter content, and levels of neurotransmission-related genes being the endpoints. 6-PPDQ exposure at 0.1-10 µg/L significantly reduced locomotion behavior, and that at 1-10 µg/L decreased sensory perception behavior in nematodes. Moreover, 6-PPDQ exposure at 10 µg/L notably induced damage to the development of dopaminergic, glutamatergic, serotonergic, and GABAergic neurons. Importantly, nematodes with chronic 6-PPDQ exposure at 10 µg/L were confirmed to suffer obviously decreased dopamine, serotonin, glutamate, dopamine, and GABA contents and altered neurotransmission-related gene expression. Meanwhile, the potential binding sites of 6-PPDQ and neurotransmitter synthesis-related proteins were further shown by molecular docking method. Lastly, Pearson's correlation analysis showed that locomotion behavior and sensory perception behavior were positively correlated with the dopaminergic, serotonergic, glutamatergic, and GABAergic neurotransmission. Consequently, 6-PPDQ exposure disturbed neurotransmitter transmission, while such changed molecular foundation for neurotransmitter transmission was related to 6-PPDQ toxicity induction. The present work sheds new lights on the mechanisms of 6-PPDQ and its possible neurotoxicity to organisms at environmentally relevant concentrations.

Improving nutrients removal of Anaerobic-Anoxic-Oxic process via inhibiting partial anaerobic mixture with nitrite in side-stream tanks: Role of nitric oxide.

A side-stream tank which was in parallel with the anoxic tank was used to improve the performance of an Anaerobic-Anoxic-Oxic process. The partial mixtures from the anaerobic tank were injected into the side-stream tank with the initial nitrite nitrogen (NO2--N) concentrations of 10 mg/L and 20 mg/L. When the initial NO2--N concentration in the tank was 20 mg/L, total nitrogen and total phosphorus removal efficiencies of the A2/O process increased from 72% and 48% to 90% and 89%, respectively. 2.23 mg/L of nitric oxide (NO) were observed in the side-stream tank. The abundance of Nitrosomonas sp. and Nitrospira sp. were varied from 0.98% and 6.13% to 2.04% and 1.13%, respectively. The abundances of Pseudomonas sp. and Acinetobacter sp. were increased from 0.81% and 0.74% to 6.69% and 5.48%, respectively. NO plays an important role for improving the nutrients removal of the A2/O process in the side-stream nitrite-enhanced strategy.

Metabolic regulation of α-Ketoglutarate associated with heat tolerance in perennial ryegrass.

α-Ketoglutarate (AKG) is a key intermediate metabolite in the tricarboxylic acid cycle of respiration and a precursor for glutamate, playing important roles in regulating plant growth and stress tolerance. The objectives of this study were to examine effects of AKG on heat tolerance characterized by leaf senescence in a cool-season grass species by foliar application and to determine major metabolites and associated metabolic pathways regulated by AKG for its effects on heat tolerance. Perennial ryegrass (Lolium perenne L.) plants were exposed to heat stress (35/30 °C, day/night) or optimal temperature (25/20 °C, day/night, non-stress control) in controlled-environment growth chambers. The solution containing AKG (5 mM) was applied to leaves by spraying 7 d prior to the initiation of heat stress and every 7 d during the heat stress period. Exogenous application of AKG enhanced heat tolerance in perennial ryegrass, as manifested by significant increases in leaf chlorophyll content, photochemical efficiency, and membrane stability, as well as activities of antioxidant enzymes for H2O2 scavenging in AKG-treated plants relative to untreated control plants exposed to heat stress. Metabolic profiling and pathway analysis demonstrated that exogenous AKG application enhanced metabolite accumulation in four major metabolic pathways, including antioxidant metabolism, amino acid metabolism, glycolysis and tricarboxylic acid cycle of respiration, and pyrimidine metabolism, contributing to AKG-improved heat tolerance in perennial ryegrass.

Mechanisms of NO and N2O production by enriched nitrifying sludge in a sequencing batch reactor: Effects of hydroxylamine.

NO and N2O as important greenhouse gases andtheir production mechanisms during nitrification are not completely understood. This study aimed to analyze the effect of hydroxylamine (NH2OH) on NO and N2O produced by nitrifying bacteria from activated sludge in a sequencing batch reactor (SBR). Experimental results showed that when nitrite (NO2-) accumulated during aerobic ammonia (NH4+) oxidation, N2O was the main product. The total amount of NO and N2O produced by NH2OH oxidation was positively correlated with dissolved oxygen (DO) levels. The imbalance of NH4+ oxidation caused by NH2OH addition was more conducive to the generation of NO and N2O under high DO conditions. When NH2OH was added into the reactor with NO2- as the substrate, the production of NO and N2O under high DO levels was mainly related to NH2OH oxidation. Under low DO conditions, NO and N2O from the biotic/abiotic hybrid pathways were more significant in the reactor of the coexistence of NO2- and NH2OH, which could be mainly caused by the pathways of nitrifier denitrification and abiotic reaction. Besides, limited amount of NO and N2O was generated by heterotrophic denitrification pathway during autotrophic nitrification. The implications for the above results are important for understanding the production of NO and N2O under NH2OH stress in nitrifying sludge reactor.

Comparison of inhibitory roles on nitrite-oxidizing bacteria by hydroxylamine and hydrazine during the establishment of partial nitrification.

The inhibitory roles of hydroxylamine (NH2OH) and hydrazine (N2H4) on nitrite-oxidizing bacteria were investigated in a comparative study. The results showed that nitrite accumulation was achieved by adding 5 mg-N/L NH2OH or N2H4 to two parallel sequencing batch reactors, with nitrite accumulation rate reaching 95.83% and 86.58% within 15 days after adopting aeration time control, respectively. Correspondingly, the maximum level of NO in typical cycles caused by NH2OH addition was 0.18 mg-N/L, which was higher than obtained for N2H4. NH2OH or N2H4 showed strong inhibition on Nitrospira and promoted the enrichment of Nitrosomonas, with the effects of NH2OH being more significant. However, nitritation began to deteriorate after the cessation of inhibitors addition. In conclusion, NH2OH was a better inhibitor than N2H4 for Nitrospira. The inhibitory role of NH2OH was primarily related to NO toxicity, while for N2H4 it was attributed to its own toxicity, with NO playing a smaller role.

Metabolic and Physiological Regulation of Aspartic Acid-Mediated Enhancement of Heat Stress Tolerance in Perennial Ryegrass.

Aspartate is the most critical amino acid in the aspartate metabolic pathway, which is associated with multiple metabolic pathways, such as protein synthesis, nucleotide metabolism, TCA cycle, glycolysis, and hormone biosynthesis. Aspartate also plays an important role in plant resistance to abiotic stress, such as cold stress, drought stress, salt stress or heavy metal stress. This study found that the chlorophyll content and antioxidant active enzyme content (SOD, CAT, POD and APX) of perennial ryegrass treated with 2 mM aspartate were significantly higher than those treated with water under heat stress. The electrolyte leakage rate, MDA content and peroxide levels (O2- and H2O2) of perennial ryegrass treated with aspartate were significantly lower than those of perennial ryegrass treated with water, indicating that exogenous aspartate increases the content of chlorophyll, maintain the integrity of cell membrane system, and enhances SOD-CAT antioxidant pathway to eliminate the oxidative damage caused by ROS in perennial ryegrass under heat stress. Furthermore, exogenous aspartate could enhance the TCA cycle, the metabolism of the amino acids related to the TCA cycle, and pyrimidine metabolism to enhance the heat tolerance of perennial ryegrass.

LpNOL-knockdown suppression of heat-induced leaf senescence in perennial ryegrass involving regulation of amino acid and organic acid metabolism.

The nonyellow COLORING 1-like gene (NOL) is known for its roles in accelerating leaf senescence, but the underlying metabolic mechanisms for heat-induced leaf senescence remain unclear. The objectives of this study were to identify metabolites and associated metabolic pathways regulated by knockdown of NOL in perennial ryegrass (Lolium perenne) and to determine the metabolic mechanisms of NOL controlling heat-induced leaf senescence. Wild-type (WT; cv. "Pinnacle") and two lines (Noli-1 and Noli-2) of perennial ryegrass with LpNOL knockdown were exposed to heat stress at 35/33°C (day/night) or nonstress control temperatures at 25/22°C (day/night) for 30 days in growth chambers. Leaf electrolyte leakage, chlorophyll (Chl) content, photochemical efficiency (Fv /Fm ), and net photosynthetic rate (Pn) were measured as physiological indicators of leaf senescence, while gas chromatography-mass spectrometry was performed to identify metabolites regulated by LpNOL. Knockdown of LpNOL suppressed heat-induced leaf senescence and produced a stay-green phenotype in perennial ryegrass, as manifested by increased Chl content, photochemical efficiency, net photosynthetic rate, and cell membrane stability in Noli-1 and Noli-2. Five metabolites (valine, malic acid, threonic acid, shikimic acid, chlorogenic acid) were uniquely upregulated in LpNOL plants exposed to heat stress, and six metabolites (aspartic acid, glutamic acid, 5-oxoproline, phenylalanine, proline, tartaric acid) exhibited more pronounced increases in their content in LpNOL plants than the WT. LpNOL could regulate heat-induced leaf senescence in perennial ryegrass through metabolic reprogramming in the pathways of respiration, secondary metabolism, antioxidant metabolism, and protein synthesis.

Improvement of the performance of simultaneous nitrification denitrification and phosphorus removal (SNDPR) system by nitrite stress.

This study investigated a new way to improve the performance of simultaneous nitrification denitrification and phosphorus removal (SNDPR) system by regularly changing the anaerobic/micro-aerobic/anoxic mode to the anaerobic/anoxic mode with 30 mg/L of nitrite dosing. The results indicated that the removal efficiency of total inorganic nitrogen and PO43--P was improved from 75.44% and 85.14% to 98.89% and 98.17%, respectively. And the good performance of the SNDPR showed a long-time sustainability when the C/N ratio was 5. The results of microbial community illustrated that the abundance of the main nitrite-oxidizing bacteria (NOB), Nitrospira sp., dropped from 5.71% to 0.85% and the abundance of denitrifying polyphosphate-accumulating organisms (DPAOs), Pseudomonas sp. and Acinetobacter sp., increased by 5 times after nitrite stress. The high level of nitric oxide (NO) and free nitrite acid produced by addition of nitrite strongly suppressed the undesired organisms NOB and ordinary heterotrophic denitrifying organisms, and promoted the enrichment of DPAOs. The NO accumulated in the nitrite denitrification process could inhibit NOB and promote AOB. This study revealed that NO plays an important role in regulating the microbial community in the SNDPR system.

The role of hydroxylamine in promoting conversion from complete nitrification to partial nitrification: NO toxicity inhibition and its characteristics.

This study investigated a strategy for hydroxylamine (NH2OH) addition for promoting the conversion of complete nitrification to partial nitrification in a sequencing batch reactor (SBR). The results showed that continuous dosing of 5 mg-N/L NH2OH into a complete nitrification reactor for 16 days led to an increase in the nitrite accumulation ratio (NAR) from 0.22% to 95.08% and a significant enhancement in the accumulation of NO and N2O in the liquid. The maximum concentration of NO in each cycle rose with the increase of NAR during NH2OH addition. With the stopping of NH2OH addition, the partial nitrification disappeared progressively in 21 days. The analysis for microbial community showed that Nitrospira was the main NOB and its relative abundance decreased with NH2OH addition and recovered after the cessation of NH2OH addition. Accordingly, NH2OH has a significant and reversible inhibition on Nitrospira and its essence might be related to NO toxicity.

Cell wall polysaccharides from pathogenic fungi for diagnosis of fungal infectious disease.

Invasive fungal diseases are associated with significant morbidity and mortality, particularly in immunocompromised individuals. Early and accurate diagnosis is crucial for effective treatment. Despite traditional methods such as microbiological culture, histopathology, radiology and direct microscopy are available, antigen/antibody-based diagnostics are emerging for diagnosis of invasive fungal infections (IFI). Fungal cell wall is a unique structure composed of polysaccharides that are well correlated with fungal burden during fungal infections. Based on this feature, cell wall polysaccharides have been explored as antigens in IFIs diagnostics such as the galactomannan assay, mannan test, ß-glucan assay and cryptococcal CrAg test. Herein, we provide an overview on the cell wall polysaccharides from three opportunistic pathogens: Aspergillus fumigatus, Candida albicans and Cryptococcus neoformans, and their applications for IFIs diagnosis. The clinical outcome of newly developed cell wall polysaccharides-based diagnostics is also discussed.