The azole biocide climbazole induces oxidative stress, inflammation, and apoptosis in fish gut.

Climbazole is an azole biocide that has been widely used in formulations of personal care products. Climbazole can cause developmental toxicity and endocrine disruption as well as gut disturbance in aquatic organisms. However, the mechanisms behind gut toxicity induced by climbazole still remain largely unclear in fish. Here, we evaluate the gut effects by exposing grass carp (Ctenopharyngodon idella) to climbazole at levels ranging from 0.2 to 20 µg/L for 42 days by evaluating gene transcription and expression, biochemical analyses, correlation network analysis, and molecular docking. Results showed that climbazole exposure increased cyp1a mRNA expression and ROS level in the three treatment groups. Climbazole also inhibited Nrf2 and Keap1 transcripts as well as proteins, and suppressed the transcript levels of their subordinate antioxidant molecules (cat, sod, and ho-1), increasing oxidative stress. Additionally, climbazole enhanced NF-κB and iκBα transcripts and proteins, and the transcripts of NF-κB downstream pro-inflammatory factors (tnfα, and il-1ß/6/8), leading to inflammation. Climbazole increased pro-apoptosis-related genes (fadd, bad1, and caspase3), and decreased anti-apoptosis-associated genes (bcl2, and bcl-xl), suggesting a direct reaction to apoptosis. The molecular docking data showed that climbazole could form stable hydrogen bonds with CYP1A. Mechanistically, our findings suggested that climbazole can induce inflammation and oxidative stress through CYP450s/ROS/Nrf2/NF-κB pathways, resulting in cell apoptosis in the gut of grass carp.

Systematic Identification, Fragmentation Pattern, And Metabolic Pathways of Hyperoside in Rat Plasma, Urine, And Feces by UPLC-Q-Exactive Orbitrap MS.

Hyperoside is a natural flavonol glycoside, which has antioxidation, antitumor, and anticancer activities together with other healthy effects like improving cardiovascular function, protecting the liver, and regulating the immune system. It is a popular compound used in the traditional Chinese medicine and different studies on hyperoside are present in the literature. However, studies on the metabolism of hyperoside in vivo were not comprehensive. In this study, UPLC-Q-Exactive Orbitrap MS technology was used to establish a rapid and comprehensive analysis strategy to explore the metabolites and metabolic process of hyperoside in rats. The metabolites of hyperoside were systematically identified in rat plasma, urine, and feces. According to the hyperoside standard substance and relevant works of literature, a total of 33 metabolites were identified, including 16 in plasma, 31 in urine, and 14 in feces. Among them, the metabolites quercetin and dihydroquercetin were unambiguously confirmed by comparison with standard substances. In addition, 13 metabolites had not been reported in hyperoside metabolism-related articles at present. The metabolic reactions of hyperoside in vivo were further explored, including phase I metabolism (hydroxylation, dehydroxylation, glycoside hydrolysis, hydrogenation, and hydration) and phase II metabolism (methylation, acetylation, sulfation, and glucuronide conjugation). The fragment ions of hyperoside and its metabolites were usually produced by glucoside bond hydrolysis, the neutral loss of (CO + OH), COH, CO, O, and Retro-Diels Alder (RDA) cleavage. In conclusion, this study comprehensively characterized the metabolism of hyperoside in rats, providing a basis for exploring its various biological activities.

High-Throughput Untargeted Serum Metabolomics Analysis of Hyperuricemia Patients by UPLC-Q-TOF/MS.

Hyperuricemia (HUA) as a metabolic disease is closely associated with metabolic disorders. The etiology and pathogenesis of HUA are not fully understood, so there is no radical cure so far. Metabolomics, a specialized study of endogenous small molecule substances, has become a powerful tool for metabolic pathway analysis of selected differential metabolites, which is helpful for initially revealing possible development mechanisms of various human diseases. Twenty HUA patients and 20 healthy individuals participated in the experiment, and ultrahigh performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry (UPLC-Q-TOF/MS) was employed to investigate serum samples to find differential metabolites. The statistical techniques used were principal component analysis and orthogonal partial least-squares discriminant analysis. The differences in metabolomics results of samples after pretreatment with different solvents were compared, 38, 20, 26, 28, 33, 50, and 40 potential differential metabolites were found, respectively, in HUA patient samples, and each group involved different metabolic pathways. Repetitive metabolites were removed, 138 differential metabolites in HUA serum were integrated for analysis, and the human body was affected by 7 metabolic pathways of glycerophospholipid metabolism, sphingolipid metabolism, arachidonic acid metabolism, linoleic acid metabolism, phenylalanine metabolism, phenylalanine, tyrosine and tryptophan biosynthesis, and α-linolenic acid metabolism. In this work, the metabolomics approach based on UPLC-Q-TOF/MS was employed to investigate serum metabolic changes in HUA patients, 138 potential differential metabolites related to HUA were identified, which provided associations of lipids, amino acids, fatty acids, organic acids, and nucleosides profiles of HUA individuals. Metabolic pathways involved in glycerophospholipid metabolism, sphingolipid metabolism, arachidonic acid metabolism, linoleic acid metabolism, phenylalanine metabolism, phenylalanine, tyrosine and tryptophan biosynthesis, and a-linolenic acid metabolism shed light on the understanding of the etiology and pathogenesis process of HUA.

Lipidomics study of the therapeutic mechanism of Plantaginis Semen in potassium oxonate-induced hyperuricemia rat.

BACKGROUND: Plantaginis Semen has been widely used as folk medicine and health care food against hyperuricemia (HUA) and gout, but its pharmacological mechanism remains unclear. This study investigated the therapeutic mechanism of Plantaginis Semen extract on potassium oxonate -induced HUA rats based on a lipidomics approach. METHODS: A model of HUA was established by potassium oxonate intragastric administration. 42 Sprague-Dawley (SD) male rats were randomly divided into the control group, model group, benzbromarone group (10 mg/kg) and three Plantaginis Semen groups (n = 7). The Plantaginis Semen groups were treated orally with Plantaginis Semen, 0.9375, 1.875  or 3.75 g/kg for 28 days. The levels of serum uric acid (UA), creatinine (Cr), triacylglycerol (TG) and tumor necrosis factor-α (TNF-α) were  measured using enzyme-linked immunosorbent assay kits. Ultra performance liquid chromatography quadrupole time of flight mass spectrometry (UPLC-Q-TOF/MS) was used for the serum lipidomics analysis, multivariate statistical analysis and independent samples t-test were carried out for the pattern recognition and characteristic metabolites identification. The relative levels of critical regulatory factors were determined by quantitative real-time polymerase chain reaction (RT-qPCR). RESULTS: Compared with the model group, the levels of serum UA, Cr, TG and TNF-α were significantly (p < 0.05) decreased in benzbromarone and three Plantaginis Semen groups. With lipidomics analysis, significant lipid metabolic perturbations were observed in HUA rats, 13 metabolites were identified as potential biomarkers and glycerophospholipid metabolism pathway was  most affected. These perturbations  were partially restored via treatment of benzbromarone and Plantaginis Semen. Additionally, the mRNA expression levels of urate anion transporter 1 (URAT1) and phosphatidylinositol 3-kinase/protein kinases B (PI3K/Akt) were significantly decreased (p < 0.01) after treatment with benzbromarone and high dose of Plantaginis Semen. CONCLUSIONS: Plantaginis Semen had significant effects on anti-HUA, anti-inflammatory and renal protection. It attenuated potassium oxonate-induced HUA through regulation of lipid metabolism disorder.

Nano-selenium controlled cadmium accumulation and improved photosynthesis in indica rice cultivated in lead and cadmium combined paddy soils.

Selenium nanoparticles (Se NPs) are less toxic and more biocompatible than selenite or selenate. However, studies involving spraying with Se NPs for reducing accumulation of cadmium (Cd) and lead (Pb) in rice grains have been rarely reported as yet. Herein, indica rice seedlings cultivated in Cd+Pb-spiked paddy soils (denoted as positive control) were sprayed with Se NPs sols for four times from tillering to booting stage. Compared to positive control, 50-100 µmol/L Se NPs downregulated Cd transporters-related genes such as OsLCT1, OsHMA2 and OsCCX2 in leaves and OsLCT1, OsPCR1 and OsCCX2 genes in node I at filling stage. Meanwhile, Se-binding protein 1 was distinctly elevated, involving the repression of Cd and Pb transportation to rice grains. Se NPs also differentially improved RuBP carboxylase and chlorophylls especially some key genes and proteins involving photosynthetic system. Besides, 25-50 µmol/L Se NPs diminished reactive oxygen species overproduction from NADPH oxidases whereas boosted glutathione peroxidase, reducing protein carbonylation in rice seedlings. However, the antioxidant isozymes and oxidatively modified proteins were slightly rebounded at 100 µmol/L. Se contents were noticeably elevated and confirmed to exist as selenomethionine in the rice grains following all the treatments by Se NPs. Thus, the optimal dosage of Se NPs for foliar application is 50 µmol/L, which significantly decreased Cd accumulation, improved photosynthesis and Se enrichment whereas caused no distinct reduction of Pb in the grains. Thus, an appropriate dosage of Se NPs can be conducted to decrease Cd accumulation, improve photosynthesis, and organic Se contents in rice grains.

Effects and mechanisms of foliar application of silicon and selenium composite sols on diminishing cadmium and lead translocation and affiliated physiological and biochemical responses in hybrid rice (Oryza sativa L.) exposed to cadmium and lead.

Currently, exploring effective measures to reduce multiple toxic metals accumulation in rice grains is an urgent issue to be tackled. Pot experiments were thus conducted to explore the effects and mechanisms of foliar spraying with composite sols of silicon (Si) and selenium (Se) during tillering to booting stage on diminishing cadmium (Cd) and lead (Pb) translocation to rice grains and affiliated physiological and biochemical responses in rice seedlings grown in Cd + Pb-polluted soils (positive control). Results showed that Cd and Pb contents in leaves or grains were distinctly below the positive control by the sols. Compared to the positive control, transcriptions of Cd transporter-related genes including OsLCT1, OsCCX2, OsHMA2 and OsPCR1 genes in leaves, and OsLCT1, OsCCX2, TaCNR2 and OSPCR1 in peduncles were downregulated by the increasing sols. Meanwhile, Se-binding protein 1 was evidently upregulated, together to retard Cd and Pb translocation to rice grains. The sols not only upregulated transcriptions of Lhcb1, RbcL, and OsBTF3 genes and production of psbA, Lhcb1 and RbcL proteins, but also increased the chlorophylls contents and RuBP carboxylase activities in the leaves, improving photosynthesis. The sols restrained ROS production from NADPH oxidases, but activated glutathione peroxidase, alleviating oxidative stress and damage. Additionally, Se was significantly enriched and was existed as selenomethionine in the rice grains. However, Pb transporter-related genes remain to be specified. Thus, the composite sols have potential to reduce Cd and Pb accumulation, mitigate oxidative damage, and promote photosynthesis and organic Se enrichment in rice plants under Cd and Pb combined pollution.

Subchronic effects of dietary selenium yeast and selenite on growth performance and the immune and antioxidant systems in Nile tilapia Oreochromis niloticus.

Selenium is an essential element but toxic at high levels in animals. The effects of Se on growth performance and the immune system in Nile tilapia remain inconclusive. In this study, Nile tilapia Oreochromis niloticus was fed on selenium yeast (Se(Y))- and selenite (Se(IV))-enriched feed at 0, 3, 6, and 12 µg/g (dry wt) for 45 and 90 d. The growth, bioaccumulation, biochemical markers related to antioxidant, immunological, nervous and digestive systems were evaluated in various fish tissues (liver, intestine, kidney, muscle, brain, spleen, gills). The results showed that the accumulation of Se(Y) was 1.3-2 folds of Se(IV) in most tissues. The growth of tilapia was enhanced by both Se(Y) and Se(IV) at 3 µg/g after 90 d, with Se(Y) better than Se(IV) in tilapia feed. After 45 d, the levels of lipid peroxidation, the activity of the antioxidant enzymes, and the transcriptional levels of the immune related genes (IL-1ß, IFN-γ and TNF-α) and stress proteins (HSP70 and MT) were enhanced in all treatments, except that of MT in the 12 µg/g Se(Y) group. In addition, both Se species inhibited the activity of acetylcholinesterase (AChE) in the brain and one digestive enzyme α-glucosidase (α-Glu) in the intestine at 12 µg/g. However, after 90 d, the effects on most biochemical markers were less pronounced, implying a possible acclimation after prolonged duration. The results demonstrate Se is beneficial to O. niloticus at low levels and toxic at elevated levels. The immunostimulation by Se might be greatly weakened after long term feeding Se-enriched feed. This study helps to better understand the effects of Se on the antioxidant and immune systems and to establish the optimal Se levels in the feed and duration for O. niloticus.

The role of the freshwater oligochaete Limnodrilus hoffmeisteri in the distribution of Se in a water/sediment microcosm.

Selenite(IV) and selenate(VI) are the major species of Se in the seleniferous aquatic ecosystem. The redistribution of Se in the water/sediment microcosm by bioturbation remains largely unknown. In this study, the redistribution of Se in the water/sediment microcosm by the benthic oligochaete Limnodrilus hoffmeisteri was assessed. The worms were exposed to 2-40 µg/g dry weight of Se(IV) or Se(VI) in the sediment (diet) for 2 months. The changes in the Se levels in different compartments of the microcosm (sediment, overlying water, and worms) were quantified after 2 weeks and 2 months. The subcellular distribution of Se in the worms were also evaluated. Finally, the volatilization of Se from the two Se sources was estimated. The results showed that Se concentration in the overlying water and Se bioaccumulation in the worms were increased with Se levels in the sediments. Approximately 1.6-9.8% of Se was volatilized in the absence of the worms and was intensified in the presence of the worms (2.1-25.7%). The subcellular distribution witnessed high levels of Se in the cell debris (>60%). Se(IV) and Se(VI) differ in their bioaccumulation, redistribution and the effects on the growth of the worms. Our results suggest that the bioturbation by benthos play an essential role in the redistribution of Se in the water/sediment microcosm.

Transcriptional Activation of Anthocyanin Biosynthesis in Developing Fruit of Blueberries ( Vaccinium corymbosum L.) by Preharvest and Postharvest UV Irradiation.

The effect and mechanism of preharvest and postharvest ultraviolet (UV) irradiation on anthocyanin biosynthesis during blueberry development were investigated. The results showed that preharvest UV-B,C and postharvest UV-A,B,C irradiation significantly promoted anthocyanin biosynthesis and the transcripts of late biosynthetic genes (LBG) VcDFR, VcANS, VcUFGT, and VcMYB transcription factor as well as DFR and UFGT activities in anthocyanin pathway in a UV wavelength- and developmental stage-dependent manner. VcMYB expression was positively correlated with that of VcANS and VcUFGT and coincided with anthocyanin biosynthesis responding to the UV radiation. Sugar decreased during postharvest but increased during preharvest UV radiation in mature fruit. Our results indicate that UV-responsive production of anthocyanins is mainly caused by the activation of anthocyanin downstream pathway genes, which could be upregulated by VcMYB. Furthermore, different potential response mechanisms may exist between preharvest and postharvest UV radiation in blueberries, involving a systemic response in living plants and a nonsystemic response in postharvest fruit.