Carcinogenic Risk of 2,6-Di-tert-Butylphenol and Its Quinone Metabolite 2,6-DTBQ Through Their Interruption of RARß: In Vivo, In Vitro, and In Silico Investigations.

Thousands of contaminants are used worldwide and eventually released into the environment, presenting a challenge of health risk assessment. The identification of key toxic pathways and characterization of interactions with target biomacromolecules are essential for health risk assessments. The adverse outcome pathway (AOP) incorporates toxic mechanisms into health risk assessment by emphasizing the relationship among molecular initiating events (MIEs), key events (KEs), and adverse outcome (AO). Herein, we attempted the use of AOP to decipher the toxic effects of 2,6-di-tert-butylphenol (2,6-DTBP) and its para-quinone metabolite 2,6-di-tert-butyl-1,4-benzoquinone (2,6-DTBQ) based on integrated transcriptomics, molecular modeling, and cell-based assays. Through transcriptomics and quantitative real-time PCR validation, we identified retinoic acid receptor ß (RARß) as the key target biomacromolecule. The epigenetic analysis and molecular modeling revealed RARß interference as one MIE, including DNA methylation and conformational changes. In vitro assays extended subsequent KEs, including altered protein expression of p-Erk1/2 and COX-2, and promoted cancer cell H4IIE proliferation and metastasis. These toxic effects altogether led to carcinogenic risk as the AO of 2,6-DTBP and 2,6-DTBQ, in line with chemical carcinogenesis identified from transcriptome profiling. Overall, our simplified AOP network of 2,6-DTBP and 2,6-DTBQ facilitates relevant health risk assessment.

Investigation of phytochemical composition and metabolite profiling in vivo of Beta vulgaris L.

RATIONALE: Beta vulgaris L. has attracted increasing attention because of its broad application. The root of B. vulgaris L. (beetroot) possesses many excellent biological properties such as antianemic, anti-inflammatory, antihypertensive, antioxidant, anticarcinogenic, antipyretic, antibacterial, detoxicant, and diuretic. The chemical constituents of beetroot play a major role in the research on beetroot application and development. At present, no systematic identification study that focuses on the chemical constituents of beetroot has been reported. METHODS: This study investigated a three-step strategy comprising phytochemical profiling, prototype profiling, and metabolism of its correlative metabolites in vivo using ultra-performance liquid chromatography tandem quadruple time-of-flight mass spectrometry (UPLC-QTOF-MS/MS). RESULTS: UPLC-QTOF-MS/MS technique proved to be a rapid, sensitive, and reliable method for monitoring the specific ingredients as well as the whole chemical constituents in beetroot. In sum, 95 phytochemical compounds, 51 prototype compounds, and 37 derived metabolites in vivo were found in beetroot. CONCLUSIONS: The main metabolic pathways were sulfonation, glucuronidation, methylation/sulfonation, and methylation. The present findings provided the phytochemical basis both in vitro and in vivo for future application.

Organ-Specific Analysis of Morus alba Using a Gel-Free/Label-Free Proteomic Technique.

Morus alba is an important medicinal plant that is used to treat human diseases. The leaf, branch, and root of Morus can be applied as antidiabetic, antioxidant, and anti-inflammatory medicines, respectively. To explore the molecular mechanisms underlying the various pharmacological functions within different parts of Morus, organ-specific proteomics were performed. Protein profiles of the Morus leaf, branch, and root were determined using a gel-free/label-free proteomic technique. In the Morus leaf, branch, and root, a total of 492, 414, and 355 proteins were identified, respectively, including 84 common proteins. In leaf, the main function was related to protein degradation, photosynthesis, and redox ascorbate/glutathione metabolism. In branch, the main function was related to protein synthesis/degradation, stress, and redox ascorbate/glutathione metabolism. In root, the main function was related to protein synthesis/degradation, stress, and cell wall. Additionally, organ-specific metabolites and antioxidant activities were analyzed. These results revealed that flavonoids were highly accumulated in Morus root compared with the branch and leaf. Accordingly, two root-specific proteins named chalcone flavanone isomerase and flavonoid 3,5-hydroxylase were accumulated in the flavonoid pathway. Consistent with this finding, the content of the total flavonoids was higher in root compared to those detected in branch and leaf. These results suggest that the flavonoids in Morus root might be responsible for its biological activity and the root is the main part for flavonoid biosynthesis in Morus.

Bioactive Penicipyrrodiether A, an Adduct of GKK1032 Analogue and Phenol A Derivative, from a Marine-Sourced Fungus Penicillium sp. ZZ380.

Penicipyrrodiether A, an adduct of GKK1032 analogue and phenol A derivative, was isolated from a culture of marine-associated fungus Penicillium sp. ZZ380 and represents the first example of this type of fungal metabolite. Its structure was elucidated by extensive spectroscopic analyses, including 1D- and 2D-NMR, HRESIMS, MS/MS, and electronic circular dichroism calculation as well as single-crystal X-ray diffraction. Penicipyrrodiether A showed antibacterial activity in inhibiting the growth of methicillin-resistant Staphylococcus aureus with a MIC value of 5.0 µg/mL. Its plausible pathway for biosynthesis has been proposed.

Metabolic Susceptibility of 2-Chlorothioxanthone and Its Toxic Effects on mRNA and Protein Expression and Activities of Human CYP1A2 and CYP3A4 Enzymes.

Thioxanthones (TXs) are photoinitiators widely used in UV curable resins and food packaging, and their residues have been frequently detected in human bodies. Our current understanding of the susceptibility of residual TXs to metabolism and their effects on human health is very limited. The in vitro metabolism of TXs and its toxic effects on cytochrome P450 (CYP) (the key xenobiotic metabolizing enzymes) were examined in this study. 2-Chlorothioxanthone (2-Cl-TX) significantly inhibited the enzymatic activities of CYP1A2 and CYP3A4 with IC50 of 8.36 and 0.86 µM, respectively. The exposure to 2-Cl-TX at 2.5 µM up-regulated the mRNA expression of CYP1A2 and CYP3A4 in human hepatocellular carcinoma cells to 3.03-fold and 2.02-fold, respectively. 2-Cl-TX at 2.5 µM caused 2.19-fold and 1.98-fold overexpression of CYP1A2 and CYP3A4, respectively. In vitro studies revealed that 2-Cl-TX was biotransformed into two metabolites through the sulfoxidation of the sulfur atom, or via the hydroxylation of aromatic carbon. Results from this study, including the metabolic susceptibility of residual 2-Cl-TX, the proposed metabolites and the significant toxic effect on the activities, mRNA, and protein expression of CYP1A2 and CYP3A4, are vital to the human health and safety risk assessment from this ubiquitous xenobiotic.

Single standard substance for the determination of nine volatile components in the distillate of Fructus Gardeniae and Radix Curcumae (an intermediate of Xingnaojing Injection).

Xingnaojing Injection is a traditional Chinese medicine extensively used for stroke and cerebral ischemia. For better in-process quality control of Xingnaojing Injection, a method for the analysis of its intermediate (i.e., the distillate of Fructus Gardeniae and Radix Curcumae) is needed to monitor and optimize the hydrodistillation extraction process. In this work, nine major volatile components in the intermediate were identified: isophorone, 4-methylene-isophorone, curcumenone, curcumenol, curdione, curzerenone, furanodienone, curcumol, and germacrone. A quantitative analysis of multi-component with a single-marker method based on high-performance liquid chromatography with diode array detection was developed for the simultaneous determination of the nine components. In this method, only curdione was needed as the reference substance, and the other eight components were determined using their relative correction factors to curdione. In the method validation, good linearity (r > 0.9999), sensitivity, repeatability, and accuracy (recoveries within 95.3-105.4%) were shown. The repeatability and robustness of the relative correction factors were studied with different column temperatures, flow rates, detection wavelengths, columns, and instruments. In sample analyses, consistent results between the proposed method and the external standard method were shown. The proposed method provides a comprehensive and low-cost tool for the quality assessment of the intermediate of Xingnaojing Injection.

Identification and quantitative analysis of physalin D and its metabolites in rat urine and feces by liquid chromatography with triple quadrupole time-of-flight mass spectrometry.

Physalin D is known to show extensive bioactivities. However, no excretion study has elucidated the excretion of physalin D and its metabolites. This study investigates the excretion of physalin D and its metabolites in rats. Metabolites in rat urine and feces were separated and identified by liquid chromatography with triple quadrupole time-of-flight mass spectrometry. Furthermore, a validated high-performance liquid chromatography with tandem mass spectrometry method was developed to quantify physalin D, physalin D glucuronide, and physalin D sulfate in rat feces and urine after the intragastric administration of physalin D. The analyte showed good linearity over a wide concentration range (r > 0.995), and the lower limit of quantification was 0.0532 µg/mL and 0.226 µg/g for urine and feces, respectively. Nine metabolites, including five phase I and four phase II metabolites, were identified and clarified after dosing in vivo. Only 4.0% of the gavaged dose, including physalin D and its phase II metabolites, was excreted in urine, whereas 10.8% was found in feces in the unchanged form. The results indicate that the extensive and rapid metabolism may be the main factors leading to the short half-life of physalin D. These results can provide a basis for further studies on the structural modification and pharmacology of physalin D.

Divergent kinetic and thermodynamic hydration of a porous Cu(II) coordination polymer with exclusive CO2 sorption selectivity.

Selective adsorption and separation of CO2 are of great importance for different target applications. Metal-organic frameworks (MOFs) represent a promising class of porous materials for this purpose. Here we present a unique MOF material, [Cu(tba)2]n (tba = 4-(1H-1,2,4-triazol-1-yl)benzoate), which shows high CO2 adsorption selectivity over CH4/H2/O2/Ar/N2 gases (with IAST selectivity of 41-68 at 273 K and 33-51 at 293 K). By using a critical point dryer, the CO2 molecules can be well sealed in the 1D channels of [Cu(tba)2]n to allow a single-crystal X-ray analysis, which reveals the presence of not only C(δ+)-H···O(δ-) bonds between the host framework and CO2 but also quadrupole-quadrupole (CO2(δ-)···(δ+)CO2) interactions between the CO2 molecules. Furthermore, [Cu(tba)2]n will suffer divergent kinetic and thermodynamic hydration processes to form its isostructural hydrate {[Cu(tba)2](H2O)}n and a mononuclear complex [Cu(tba)2(H2O)4] via single-crystal to single-crystal transformations.

A pair of chiral flavonolignans as novel anti-cyanobacterial allelochemicals derived from barley straw (Hordeum vulgare): characterization and comparison of their anti-cyanobacterial activities.

The inhibitory effect of barley straw (Hordeum vulgare) on cyanobacteria has been observed in many field and laboratory studies for over 30 years, although the compounds responsible for this anti-cyanobacterial effect have remained unknown. In this study, a pair of chiral flavonolignans were isolated from barley straw extract using a bioassay-guided isolation procedure against Microcystis sp. The structures of the allelopathic compounds were elucidated by NMR (nuclear magnetic resonance) and HPLC-MS (high performance liquid chromatography-mass spectrometry), and turned out to be salcolin A and B. The enantiomers differ in their anti-cyanobacterial abilities. Both enantiomers exhibited inhibitory effects on Microcystis sp., and the EC50 (concentration for 50% of maximal effect) of salcolin A and B were 6.02 × 10(-5) and 9.60 × 10(-5 ) mol l(-1) , respectively. Furthermore, the modes of actions of the enantiomers were investigated and compared at a single cell level by flow cytometry. Salcolin A was found to induce an increase on cyanobacterial intracellular ROS (reactive oxygen species) levels and to inhibit esterase activity, whereas salcolin B caused leakages of cyanobacterial cytoplasms. Thus, salcolin A was more 'algistatic', and salcolin B was more 'algicidal'. This study suggests that salcolin is the key allelochemical in barley straw's inhibitory effect on cyanobacteria and could be used as an agent in the future control of cyanobacterial harmful algae blooms.

[Identification of phenylpropanoids in ciwujia injection by HPLC-MS].

There are reports about the chemical compounds of Ciwujia herbs, but with no study report about the chemical material basis of Ciwujia injection (CWJI). In this study, LC-MS(n) and LC-Q-TOF-MS techniques were adopted for a qualitative analysis on phenylpropanoids in CWJI. The Ultmate XB-C18 column (4.6 mm x 250 mm, 5 microm) was adopted and eluted with the mobile phase of 0.5% formic acid-water and acetonitrile, with the flow rate at 0.8 mL x min(-1) and the column temperature at 20 degrees C. Based on the data of high-resolution and multi-stage MS, control products and literatures, altogether 54 phenylpropanoids were identified in Ciwujia Injection, including 34 phenylpropanoids, 16 ligans and 4 coumarins. Among them, 28 were reported for the first time in Ciwujia, and 14 compound structures were identified in comparison with the control products. The method established in this study could be used to simply and rapidly identify phenylpropanoids in CWJI. The findings provide scientific data for defining the chemical material basis of CWJI.

Image Captioning With Controllable and Adaptive Length Levels.

Image captioning is a core challenge in computer vision, attracting significant attention. Traditional methods prioritize caption quality, often overlooking style control. Our research enhances method controllability, enabling descriptions of varying detail. By integrating a length level embedding into current models, they can produce detailed or concise captions, increasing diversity. We introduce a length-level reranking transformer to correlate image and text complexity, optimizing caption length for informativeness without redundancy. Additionally, with caption length increase, computational complexity grows due to the autoregressive (AR) design of existing methods. To address this, our non-autoregressive (NAR) model maintains constant complexity regardless of caption length. We've developed a training approach that includes refinement sequence training and sequence-level knowledge distillation to close the performance gap between NAR and AR models. In testing, our models set new standards for caption quality on the MS COCO dataset and offer enhanced controllability and diversity. Our NAR model excels over AR models in these aspects and shows greater efficiency with longer captions. With advanced training techniques, our NAR's caption quality rivals that of leading AR models.

Preparation of bacterial fertilizer from biogas residue after anaerobic co-digestion of kitchen waste and residual sludge.

Azotobacter chroococcum and Bacillus subtilis were selected as fermentation strains, and biogas residue after anaerobic digestion of kitchen waste and residual sludge was used as fermentation substrate. A single factor optimization test was used to optimize the solid-state fermentation parameters of biogas residue with the number of viable bacteria and the number of spores as indexes. The results showed that the optimum inoculation conditions involved the following: 55% initial moisture content, 15% initial inoculation amount, 30 ℃, and 1:1 initial inoculation ratio for 13 days. Pot experiment showed that the prepared three kinds of bacterial fertilizers could not only effectively promote the growth of white clover, improve the composition of soil nutrients, but also change the structure of soil bacterial community, which is of great significance to the health of soil ecosystem in white clover.

The evolutionary adaptation of wood-decay macrofungi to host gymnosperms differs from that to host angiosperms.

Wood-decay macrofungi play a vital role in forest ecosystems by promoting nutrient cycling and soil structure, and their evolution is closely related to their host plants. This study investigates the potential evolutionary adaptation of wood-decay macrofungi to their host plants, focusing on whether these relationships differ between gymnosperms and angiosperms. While previous research has suggested non-random associations between specific fungi and plant deadwood, direct evidence of evolutionary adaptation has been lacking. Our study, conducted in a subtropical region, utilized metabarcoding techniques to identify deadwood species and associated fungi. We found significant evidence of evolutionary adaptation when considering all sampled species collectively. However, distinct patterns emerged when comparing angiosperms and gymnosperms: a significant evolutionary adaptation was observed of wood-decay macrofungi to angiosperms, but not to gymnosperms. This variation may be due to the longer evolutionary history and more stable species interactions of gymnosperms, as indicated by a higher modularity coefficient (r = .452), suggesting greater specialization. In contrast, angiosperms, being evolutionarily younger, displayed less stable and more coevolving interactions with fungi, reflected in a lower modularity coefficient (r = .387). Our findings provide the first direct evidence of differential evolutionary adaptation dynamics of these fungi to angiosperms versus gymnosperms, enhancing our understanding of forest ecosystem carbon cycling and resource management.

Characterization of Various Noncovalent Polyphenol-Starch Complexes and Their Prebiotic Activities during In Vitro Digestion and Fermentation.

This study explores the structural characterization of six noncovalent polyphenol-starch complexes and their prebiotic activities during in vitro digestion and fermentation. Ferulic acid, caffeic acid, gallic acid, isoquercetin, astragalin, and hyperin were complexed with sweet potato starch (SPS). The polyphenols exhibited high binding capacity (>70%) with SPS. A partial release of flavonoids from the complexes was observed via in vitro digestion, while the phenolic acids remained tightly bound. Molecular dynamics (MD) simulation revealed that polyphenols altered the spatial configuration of polysaccharides and intramolecular hydrogen bonds formed. Additionally, polyphenol-SPS complexes exerted inhibitory effects on starch digestion compared to gelatinized SPS, owing to the increase in resistant starch fraction. It revealed that the different complexes stimulated the growth of Lactobacillus rhamnosus and Bifidobacterium bifidum, while inhibiting the growth of Escherichia coli. Moreover, in vitro fermentation experiments revealed that complexes were utilized by the gut microbiota, resulting in the production of short-chain fatty acids and a decrease in pH. In addition, the polyphenol-SPS complexes altered the composition of gut microbiota by promoting the growth of beneficial bacteria and decreasing pathogenic bacteria. Polyphenol-SPS complexes exhibit great potential for use as a prebiotic and exert dual beneficial effects on gut microbiota.

[Study on rapid screening and identifying hepatotoxic compounds in toosendan fructus].

Fluorescein diacetate-labeled HepG2 cells model and flouresence automatic microscopy screening assay were used for fast screening 23 components from Toosendan Fructus, in which 5 components showed significant toxicity on HepG2 cells. The 10 compounds in the 2 components were tentatively identified with LC-MS(n), and 3 of them (meliasenin B, trichilinin D and 1-O-tigloy-1-O-debenzoylohchinal) were prepared and identified. Further experiments showed that the 3 compounds displayed dose-dependent toxicity on HepG2 cells, suggesting that these compounds in Toosendan Fructus may cause hepatotoxicity.

Dietary Xylitol Supplement Ameliorated AD-related Neuronal Injury by Regulating Glucose Metabolism Relevant Amino Acids in Mice.

BACKGROUND: Alzheimer's disease (AD) is one of the most common irreversible degenerative diseases of the central nervous system. Recent studies have found that patients with AD generally experience abnormal glucose metabolism. Xylitol is a functional sugar alcohol, which has been reported to regulate glucose metabolism. OBJECTIVE: The present study was designed to determine whether xylitol can alleviate cognitive impairment in AD mice. METHODS: In the current research, 5% xylitol was supplemented in the diet to treat APP/PS1 transgenic AD mice for 2 months. Cognitive ability was measured by the Morris water maze, and anxiety-like behaviors were examined by open-field experiment. Hippocampal cellular apoptosis and mitochondria pathway related apoptotic proteins were tested by TUNEL staining and immunoblotting, respectively. By LC-MS, plasma levels of glucose metabolism intermediates and related amino acids were evaluated. RESULTS: Results showed that xylitol could significantly ameliorate anxiety-like activity in AD mice by partially regulating expression levels of mitochondrial pathway-related apoptotic proteins. Xylitolregulated glucose metabolism may play an important role in the process. CONCLUSION: The current study suggests that xylitol may be a potential candidate for improving neuropsychiatric behavior in AD by regulating the levels of TCA cycle intermediates and related amino acids in glucose metabolism.

Structural, in vitro digestion, and fermentation characteristics of lotus leaf flavonoids.

Bioaccessibility and bioactivity of flavonoids in lotus leaves are related to their characteristics in gastrointestinal digestion and colonic fermentation. The aim of this study is to investigate the stability of lotus leaf flavonoids (LLF) in simulated gastrointestinal digestion, and its modulation on gut microbiota in vitro fermentation. Results showed that LLF mainly consisted of quercetin-3-O-galactoside, quercetin-3-O-glucuronide, quercetin-3-O-glucoside, and kaempferol-3-O-glucoside. These flavonoids kept stability with only a small fraction degraded in simulated gastric and intestinal fluids. In vitro fermentation, LLF stimulated the growth of Actinobacteria and Firmicutes, inhibited the growth of Proteobacteria, and induced the production of fermentation gases and short-chain fatty acids. Interestingly, supplementation of soluble starch significantly improved the utilization of LLF by the intestinal flora. These results revealed that LLF shaped a unique biological web with Lactobacillus and Bifidobacterium spp. as the core of the biological network, which would be more beneficial to gut health.

Oil diffusion mathematical model in oily sludge particle under supercritical water environment.

Supercritical water (SCW, T > 374.15 °C, P > 22.1 MPa) treatment can achieve volume reduction, harmless disposal, and resource utilization of oily sludge. Herein, we investigated the oil removal efficiency (ORE) and oil diffusion characteristics in oily sludge particles under SCW environment. The experimental results showed that when the treatment duration was extended from 5 min to 60 min, the particle diameter decreased from 4 mm to 2 mm, and the ORE improved considerably; however, the treatment temperature (375 °C ∼ 425 °C) had little influence. Based on these findings, an oil diffusion mechanism in oily sludge particles under SCW environment was proposed. Subsequently, a reasonable mathematical model of diffusion was developed to represent the heat and mass transfer in oily sludge particles characterized by porous, high moisture, and oil content. Finally, by analyzing the oil diffusion process in sludge particles within this model, it was found that the oil concentration in SCW and particle diameter had a considerable influence on ORE, while the effect can be ignored when the diameter < 0.2 mm. This research serves as a guide for effectively using SCW to remove oil from oily sludge.

Experimental study on supercritical water gasification of oily sludge using a continuous two-step method.

Supercritical water gasification (SCWG) technology can convert oily sludge into hydrogen-rich gas. To achieve high gasification efficiency of oily sludge with a high oil concentration under mild conditions, a two-step method involving a desorption process and a catalytic gasification process using Raney-Ni catalyst was investigated. High oil removal efficiency (99.57%) and carbon gasification efficiency (93.87%) were achieved. The lowest wastewater total organic carbon, oil content, and carbon content in the solid residues were 4.88 ppm, 0.08% and 0.88%, respectively, using a gasification temperature of 600 °C, treatment concentration of 1.11 wt%, gasification time of 70.7 s, and the optimal desorption temperature of 390 °C. The main organic carbon component in the solid residues was cellulose, which is environmentally safe. As the treatment concentration increased, the two-step method outperformed the single-step method. The mechanism for the two-step SCWG of oily sludge was revealed. In the first step, supercritical water is used in the desorption unit to achieve a high oil removal efficiency with few liquid products generated. In the second step, the Raney-Ni catalyst promotes efficient gasification of high-concentration oil at a low temperature. This research provides valuable insights into the effective SCWG of oily sludge at a low temperature.

The Tandem of Liquid Chromatography and Network Pharmacology for the Chemical Profiling of Pule'an Tablets and the Prediction of Mechanism of Action in Treating Prostatitis.

Prostatitis, a prevalent urinary tract disorder in males, has a complex etiology that leads to severe clinical discomfort. Pule'an Tablets, a classic single-component formulation primarily based on rapeseed pollen, have been clinically proven to have a beneficial therapeutic effect on both prostatitis and benign prostatic hyperplasia. However, there is currently a lack of research on the chemical composition and mechanisms of action of Pule'an Tablets in treating prostatitis. In this study, using liquid chromatography-mass spectrometry (LC-MS), a total of 53 compounds in Pule'an Tablets were identified, including flavonoids, phenylpropionamides, lipids, glucosinolates, and nucleic acids. Subsequently, through a network pharmacology analysis, potential target genes and their mechanisms of action were predicted accordingly. The results suggested that genes such as LPAR5, LPAR6, LPAR4, LPAR3, LPAR2, LPAR1, F2, ENPP2, MMP9, and TNF, along with pathways like prostate cancer, endocrine resistance, bladder cancer, and the IL-17 signaling pathway, may represent potential pathways involved in the therapeutic effects of Pule'an Tablets. This study represents the first systematic investigation into the chemical composition of Pule'an Tablets, shedding light on the potential mechanisms underlying their efficacy in treating prostatitis. These findings could serve as a valuable reference for future pharmacological research on Pule'an Tablets.