A novel three-layer module BoMYB1R1-BoMYB4b/BoMIEL1-BoDFR1 regulates anthocyanin accumulation in kale.

Anthocyanin is an important pigment responsible for plant coloration and beneficial to human health. Kale (Brassica oleracea var. acephala), a primary cool-season flowers and vegetables, is an ideal material to study anthocyanin biosynthesis and regulation mechanisms due to its anthocyanin-rich leaves. However, the underlying molecular mechanism of anthocyanin accumulation in kale remains poorly understood. Previously, we demonstrated that BoDFR1 is a key gene controlling anthocyanin biosynthesis in kale. Here, we discovered a 369-bp InDel variation in the BoDFR1 promoter between the two kale inbred lines with different pink coloration, which resulted in reduced transcriptional activity of the BoDFR1 gene in the light-pink line. With the 369-bp insertion as a bait, an R2R3-MYB repressor BoMYB4b was identified using the yeast one-hybrid screening. Knockdown of the BoMYB4b gene led to increased BoDFR1 expression and anthocyanin accumulation. An E3 ubiquitin ligase, BoMIEL1, was found to mediate the degradation of BoMYB4b, thereby promoting anthocyanin biosynthesis. Furthermore, the expression level of BoMYB4b was significantly reduced by light signals, which was attributed to the direct repression of the light-signaling factor BoMYB1R1 on the BoMYB4b promoter. Our study revealed that a novel regulatory module comprising BoMYB1R1, BoMIEL1, BoMYB4b, and BoDFR1 finely regulates anthocyanin accumulation in kale. The findings aim to establish a scientific foundation for genetic improvement of leaf color traits in kale, meanwhile, providing a reference for plant coloration studies.

Presence of benzotriazole ultraviolet stabilizers in human urine.

Health exposure to benzotriazole ultraviolet stabilizers (BUVSs) may pose diverse toxic impacts on health. Presently, the occurrence of BUVSs in human urine remains inadequately understood. This study analyzed 13 kinds of BUVSs in human urine (n = 182) from the general Chinese adult participants. Totally, nine BUVSs were measurable in these human urine samples. Among the detected BUVSs, 2-(2H-benzotriazol-2-yl)-p-cresol (UV-P) was the most predominant BUVS in the human urine, with the mean concentration of 1.6 µg/g creatinine (

Development of QuEChERS coupled with UHPLC-MS/MS for simultaneous determination of eight neonicotinoid pesticides in breast milk.

A speedy and hypersensitive method was built to detect eight neonicotinoid insecticides (neonics) in breast milk by ultrahigh-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). The breast milk was extracted with a mixture of acetonitrile and water and purified with primary secondary amine (PSA) and C18. The recovery of the method ranged from 74.3 to 105.9% with relative standard deviations (RSDs) of less than 10%, and the limit of detection ranged from 0.05 to 0.18 ng/mL. Among 32 samples obtained from women 1 month postpartum, thiamethoxam and imidacloprid were the most frequently detected neonics. Moreover, thiacloprid and imidaclothiz were not detected in any samples. The concentrations of neonics in breast milk ranged from 1.90 to 149.95 ng/mL. Considering the toxic effects on mammals and even humans, infants who are exposed to neonics through ingestion of breast milk should receive extensive attention in future studies.

The influences of perfluoroalkyl substances on the rheumatoid arthritis clinic.

BACKGROUND: The effect of environmental factors on genetically susceptible individuals is a basic link in the pathogenesis of rheumatoid arthritis. Perfluoroalkyl substances (PFASs) are a class of synthetic organic fluorine chemicals, which have been mass-produced and widely used in the past 60 years, and also have been shown to be one of the major pollutants affecting human health. The impact of fluoride on the development of Rheumatoid Arthritis (RA) is unclear. This study explored the relationship between common fluoride and clinical manifestations of rheumatoid arthritis. RESULTS: A cohort of 155 patients with RA and 145 health controls in Second Affiliated Hospital of Zhejiang University School of Medicine were investigated. Serum concentrations of all fluoride detected were higher in RA patients than in healthy controls. There were 43 male patients and 112 female patients in the RA cohort. Some of perfluoroalkyl substances (perfluorooctanoate (PFOA), perfluorononanoate (PFNA), perfluorotrdecanoate (PFTrA), perfluorooctanesulfonate (PFOS)) were correlated negatively with the Body Mass Index (BMI); some of them (PFOA, PFNA, PFTrA, PFOS, 8:2 Chlorinated polyfluorinated ether sulfonate (8:2Cl-PFESA)) were correlated positively with the Disease Activity Score 28 (DAS28); two (PFOA, PFOS) of them were correlated positively with the white blood cell count, and one (Perfluoroundecanoate (PFUnA)) of them was correlated negatively with the hemoglobin; two (Perfluorodecanoate (PFDA), PFUnA) of them were correlated negatively with the presence of interstitial lung disease. CONCLUSION: These data suggest that exposure to perfluoroalkyl substances may promote the disease activity of rheumatoid arthritis and the visceral lesions.

Effects of Bisphenol A and Bisphenol S Exposure at Low Doses on the Metabolome of Adolescent Male Sprague-Dawley Rats.

Toxic effects induced upon exposure to low-dose bisphenol A (BPA) or bisphenol S (BPS) remains controversial. In this study, metabolomics was used to examine the metabolomic perturbation arising from 28 days of exposure to BPA or BPS at 50 µg/kg bw/day in Sprague-Dawley (SD) rats. Endogenous metabolite profiling revealed a clear discrimination of metabolome in the rat plasma among BPA-treatment, BPS-treatment, and control groups. BPA exposure induced the up-regulation of 19 metabolites and down-regulation of 32 metabolites in plasma of SD rats, compared with the control. BPS exposure induced the up-regulation of 15 metabolites and the down-regulation of 33 metabolites in the plasma of SD rats, compared with the control. Joint pathway analysis suggested marked perturbations in the citrate cycle, butanoate metabolism, and alanine, aspartate, and glutamate metabolism for BPA-exposed rats as well as glycerophospholipid metabolism for BPS-exposed rats. These findings provide novel insights into associations between the metabolomic perturbation and phenotypic changes arising from BPA and BPS exposure.

Occurrence of Free-Form and Conjugated Bisphenol Analogues in Marine Organisms.

Upon exposure, most bisphenol analogues (BPs) are rapidly metabolized to BP conjugates in organisms. Monitoring studies on BPs in aquatic organisms have been mainly focused on free-form BPs. However, the relative amount of conjugated BPs in organisms is still not well known, especially in marine organisms. In this study, we collected marine organisms (13 species; n = 74), as well as seawater (n = 15), from East China Sea, and analyzed them for bisphenol A (BPA) and its eight analogues. In seawater, BPA was the predominant BP (mean ± standard deviation (SD), 18 ± 9.7 ng/L), followed by bisphenol S (BPS; 3.7 ± 2.8 ng/L), bisphenol F (BPF; 0.31 ± 0.17 ng/L), and bisphenol AF (BPAF; 0.24 ± 0.15 ng/L). The whole body of each marine organism was used for BP analysis in this study. BPA (mean 3.8 ng/g, range 1.2-7.7 ng/g) and BPS (1.5 ng/g, 0.19-6.1 ng/g) were still the predominant BPs in marine organisms without hydrolysis. After enzymatic hydrolysis treatment, mean concentrations of BPs increased 1.8 (BPS)-3.7 (BPA) times in marine organisms. Correspondingly, on average, 74, 52, 49, 48, and 45% of BPA, bisphenol Z, BPF, BPS, and BPAF were present in conjugated forms, respectively, in marine organisms. Moreover, bioaccumulation and biomagnification of BPs were evaluated using pooled marine organism samples. We first found that the mean log BAF values of BPs increased 0.091 (BPAF) to 0.31 (BPA) times in marine organisms after hydrolysis. Overall, this study first determines the amount of BPs present in conjugated forms in marine organisms, which are helpful for better understanding the occurrence of BPs in organisms.

Determination of polyhalogenated carbazoles in waters at low nanogram-per-liter concentrations with solid-phase disk extraction.

Polyhalogenated carbazoles, a class of emerging contaminants with persistence and dioxin-like toxicity, have received increasing attention in recent years. In this study, a simple, rapid, sensitive, and high throughput method based on solid-phase disk extraction and gas chromatography-mass spectrometry was described for the determination of polyhalogenated carbazoles in low nanogram-per-liter range in water samples. The proposed solid-phase disk extraction method was initially optimized, and the optimum experimental conditions found were 1 L water sample (pH 6-9) extracted and enriched by Empore 3-stn octadecyl disk at flow rate of 5 to 50 mL/min and eluted by 5 mL of acetone and 3 × 10 mL methylene dichloride. The linearity of the method ranged from 0.2 to 50 ng/L for carbazole and 11 polyhalogenated carbazoles, with correlation coefficients ranging from 0.9951 to 0.9996. The limits of detection were in the low nanogram per liter level, ranging from 0.018 to 0.12 ng/L. Finally, the optimized method was applied for determining trace levels of carbazole and 11 polyhalogenated carbazoles in tap water and seawater samples with good recovery of 86.6-112.8%. Carbazole and 3-7 polyhalogenated carbazoles were detected, and 3,6-dichlorocarbazole was the predominant congener both in tap water and seawater.

Determination of Environmental Micro(Nano)Plastics by Matrix-Assisted Laser Desorption/Ionization-Time-of-Flight Mass Spectrometry.

Micro(nano)plastics (MNPs) are widely acknowledged as global environmental threat while determination methods for MNPs are still lacking and becoming a growing concern. This study provides a novel method for MNPs identification/quantification by matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF MS). Factors affecting the measurement were optimized, including laser energy, matrix (M), analyte (A), cationization agent (C), and MAC volume ratio. Under the optimal conditions, the peaks representative of polystyrene (PS) and polyethylene terephthalate (PET) were identified, and the mass differences were consistent with the molecular weight of the corresponding oligomer. A quantitative correlation was built between normalized signal intensity and ln[polymer concentration] with a correlation coefficient above 0.96 for low-molecular-weight polymers and 0.98 for high-molecular-weight polymers. Furthermore, two types of environmental MNP samples were prepared, including aviation cup particles as fresh plastics and aged MNPs extracted from river sediment. By using MALDI-TOF MS, the PS-related MNPs (in both aviation cup and sediment) consisted of C8H8 and C16H16O oligomers, while the PET-related MNPs (only found in sediment) were identified with repeated units of C10H8O4 and C12H12O4. According to the quantitative correlation curve, the contents of PS and PET MNPs were quantified as 8.56 ± 0.04 and 28.71 ± 0.20 mg·kg-1, respectively, in the collected sediment. This study is the first attempt to propose a quantification method with the employment of MALDI-TOF MS for aged MNPs analysis in environmental samples, which can not only supply an effective method for MNP analysis but also inspire future studies on the in situ distribution and transformation of MNPs in environmental and biological samples.

Perfluoroalkyl Acids Including Isomers in Tree Barks from a Chinese Fluorochemical Manufacturing Park: Implication for Airborne Transportation.

Measurement of airborne perfluoroalkyl acids (PFAAs) is challenging, but important for understanding their atmospheric transport. Tree bark is good media for monitoring semivolatile compounds in the atmosphere. Whether it could work as an indicator of airborne PFAAs was first examined in this study. Bark and leaf samples collected around a Chinese fluorochemical manufacturing park (FMP) were analyzed for PFAAs and their branched isomers. Total PFAA concentrations (∑PFAAs) in the bark (mean, 279 ng/g dw) and leaf (250 ng/g dw) samples were comparable. ∑PFAAs in the barks collected within the boundaries of the FMP were significantly (p < 0.05) higher than those outside the FMP, and displayed a decreasing spatial trend as the distance from the FMP increased. However, such spatial difference and trend were not observed for the leaves. PFAA compositional profiles in most of the tree barks were consistent with each other, but different from those in tree leaves. These results indicated that tree barks mainly accumulated airborne PFAAs, while uptake from soil and translocation could make partial contribution to those in leaves. Perfluorooctanoate and perfluorooctanesulfonate in barks had strictly consistent isomeric compositions with their electrochemical fluorination products. Overall, these results indicated that the bark could be a good indicator of airborne PFAAs with respect to their occurrence, isomeric signature, and atmospheric transport.

Occurrence and Partitioning of Bisphenol Analogues in Adults' Blood from China.

Widespread human exposure and associated adverse health effects led to regulations on the usage of bisphenol A (BPA). Several bisphenol analogues (BPs) have been introduced as BPA alternatives in various applications. However, these BPs have been shown to exhibit similar or even stronger endocrine-disrupting activities compared with that of BPA. Currently, information on the human exposure to BPA alternatives remains limited. In this study, nine BPs were quantified in 81 pairs of plasma and red blood cell (RBC) samples from Chinese participants. In human plasma, the predominant BPs was BPA, bisphenol S (BPS), and bisphenol AF (BPAF), with the mean concentrations of 0.40, 0.15, and 0.073 ng/mL, respectively. BPA (accounting for 63% of total BPs) and BPS (18%) were the major BPs in the RBC fraction. Mass fractions in plasma (Fp) were found to be highest for BPS (mean, 0.78), followed by BPAF (0.71) and BPA (0.67), indicating strong partitioning to the plasma fraction. However, bisphenol AP was more frequently detected in the RBC fraction. Estimated total daily intake (EDI) of BPA was in the range of 0.0048-0.75 µg/kg bw/day for the participants, and adults aged >50 years had comparatively lower EDI. To our knowledge, this is the first study to assess the occurrence and partitioning of BPA alternatives in paired human plasma and RBCs from the Chinese general population.

Isomer-Specific Distribution of Perfluoroalkyl Substances in Blood.

Perfluoroalkyl substances (PFASs) such as perfluorohexanesulfonate (PFHxS), perfluorooctanoate (PFOA), perfluorooctanesulfonate (PFOS) and PFOS-precursors are routinely measured in human plasma and serum, but their relative abundance in the blood cell fraction has not been carefully examined, particularly at the isomer-specific level. Human plasma and whole blood were collected and partitioning behaviors of PFASs and their isomers between plasma and blood cells were investigated. In human samples, mass fraction in plasma (Fp) for PFASs increased among perfluoroalkyl carboxylates as the carbon chain length increased from C6 (mean 0.24) to C11 (0.87), indicating preference for the plasma fraction with increasing chain length. However, among perfluoroalkyl sulfonates, PFHxS (mean 0.87) had a slightly higher Fp than PFOS (0.85). In vitro assays with spiked Sprague-Dawley rat blood were also conducted, and the results showed that PFOS-precursors had lower Fp values than perfluoroalkyl acids, with perfluoroctanesulfonamide having the lowest Fp (mean 0.24). Consistently, linear isomers of PFOS and PFOS-precursors had lower mean Fp than their corresponding total branched isomers. Multiplying by a factor of 2 is not a reasonable method to convert from whole blood to plasma PFAS concentrations, and current ratios could be used as more accurate conversion factors.

Isomer profiles of perfluoroalkyl substances in water and soil surrounding a chinese fluorochemical manufacturing park.

Despite that China is the largest global manufacturer of perfluoroalkyl substances (PFASs), the manufacturing methods and isomer purity of these chemicals are generally unknown. Here, sampling was conducted around a major fluorochemical manufacturing park in China in 2012, including soil and water collection inside the park, including from a wastewater treatment plant (WWTP), as well as in surrounding rivers and soil (∼15 km radius). Perfluoroalkyl sulfonates (PFSAs) were lower than perfluoroalkyl carboxylates (PFCAs) in all samples, and short-chain (C4-C6) PFCAs were predominant. Perfluoroalkyl phosphonates and phosphate diesters were occasionally detected, but at low detection frequency. Branched isomers of perfluorobutanesulfonate (PFBS) are reported for the first time, accounting for 15-27% of total PFBS in water. An enrichment of isopropyl-PFOA (28%) was found in WWTP influent, suggesting its manufacturing primarily by isopropyl telomerization. More numerous branched isomers were observed for the longer C9-C13 PFCAs (e.g., C12 PFCA had 16 branched isomers), including high proportions of one major branched isomer (likely isopropyl), possibly as impurities from isopropyl-PFOA manufacturing. Overall, short-chain perfluorinated acids were the predominant PFASs being released, but PFOA was still a major chemical in use at this site, primarily from isopropyl telomerization.

Airborne trifluoroacetic acid and its fraction from the degradation of HFC-134a in Beijing, China.

Trifluoroacetic acid (TFA) has been attracting increasing attention worldwide because of its increased environmental concentrations and high aquatic toxicity. Atmospheric deposition is the major source of aquatic TFA, but only a few studies have reported either air concentrations or deposition fluxes for TFA. This is the first study to report the atmospheric concentrations of TFA in China, where an annular denuder and filter pack collection system were deployed at a highly urbanized site in Beijing. In total, 144 air samples were collected over the course of 1 year (from May 2012 to April 2013) and analyzed directly using high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) or following derivatization by gas chromatography-mass spectrometry (GC-MS). The annual mean atmospheric concentration of TFA was 1580 ± 558 pg/m(3), higher than the previously reported annual mean levels in Germany and Canada. For the first time, it was demonstrated that maximum concentrations of TFA were frequently observed in the afternoon, following a diurnal cycle and suggesting that a major source of airborne TFA is likely degradation of volatile precursors. Using a deposition model, the annual TFA deposition flux was estimated to be 619 ± 264 µg m(-2) year(-1). Nevertheless, a box model estimated that the TFA deposition flux from the degradation of HFC-134a contributed only 14% (6-33%) to the total TFA deposition flux in Beijing. Source analysis is quite important for future TFA risk predictions; therefore, future research should focus on identifying additional sources.

Sediment-seawater partitioning, bioaccumulation, and biomagnification of perfluorobutane sulfonamide in marine environment.

Environmental occurrence of perfluorobutane sulfonamide (PFBSA) has only been recently discovered. The current knowledge regarding the occurrence and environmental behaviors of PFBSA in the marine environment is still relatively limited. In this study, PFBSA and other 37 poly- and perfluoroalkyl substances were analyzed in seawater (n = 43), sediment (n = 43), and marine fish (n = 176) samples collected from East China Sea and Antarctic Ocean. PFBSA was detected in > 90% of seawater from East China Sea and Antarctic Ocean, with the concentrations of 1.0 - 19 ng/L and < LOD-228 pg/L, respectively. The field-based mean log-transformed sediment-seawater partitioning coefficients of PFBSA were 1.6 ± 0.19 L/kg dw and 1.1 ± 0.19 L/kg dw in East China Sea and Antarctic Ocean, respectively, which are lower than that of perfluorooctanoate and perfluorooctane sulfonate. This indicates its long-range transport potential in global oceans with ocean currents. The mean log-transformed bioaccumulation factor values of PFBSA determined in the multiple species of whole-body marine fishes from East China Sea and Antarctic Ocean were 2.3 L/kg ww and 2.4 L/kg ww, respectively, which are comparable to that of perfluoroheptanoate (2.3 L/kg ww) in marine fishes from East China Sea. We did not observe an obvious biomagnification or biodilution of PFBSA along the marine food chain in East China Sea or Antarctic Ocean. This study provides the first data on the environmental behaviors of PFBSA in the marine environment.

Conjugated bisphenol S metabolites in human serum and whole blood.

Studies have shown that bisphenol S (BPS) is mainly present as its conjugated metabolites in human blood. However, the distribution of conjugated BPS metabolites in different human blood matrices has not been characterized. In this study, paired human serum and whole blood samples (n = 79) were collected from Chinese participants, and were measured for the occurrence of BPS and 4 BPS metabolites. BPS was detectable in 49% of human serum (

Bisphenol S and Its Chlorinated Derivatives in Indoor Dust and Human Exposure.

Bisphenol S (BPS), an environmental endocrine disruptor, has been identified in global environmental matrices. Nevertheless, limited studies have investigated the presence of chlorinated analogues of BPS (Clx-BPSs) with potential estrogenic activities in environmental matrices. In this study, the occurrence of BPS and five types of Clx-BPSs was characterized in indoor dust (n = 178) from Hangzhou City. BPS was measurable in 94% of indoor dust samples, with an average level of 0.63 µg/g (

Occurrence and partitioning of p-phenylenediamine antioxidants and their quinone derivatives in water and sediment.

p-Phenylenediamine antioxidants (PPDs) and PPDs-derived quinones (PPDQs) may pose a threat to the river ecosystem. However, the knowledge on the occurrence and environmental behaviors of PPDs and PPDQs in the natural river environment remains unknown. In this study, we collected paired water (n = 30) and sediment samples (n = 30) from Jiaojiang River, China and analyzed them for nine PPDs and seven PPDQs. Our results showed that target PPDs and PPDQs are frequently detected in water samples, with the dominance of N-(1,3-dimethylbutyl)-N'-phenyl-p-phenylenediamine (6PPD; mean 12 ng/L, range 4.0-72 ng/L) and 6PPD-derived quinone (6PPDQ; 7.0 ng/L,

Occurrence of p-phenylenediamine antioxidants (PPDs) and PPDs-derived quinones in indoor dust.

The p-phenylenediamine antioxidants (PPDs) have been widely detected in various dust samples. Nevertheless, the knowledge on occurrence of their environmental transformation products, PPD-derived quinones (PPDQs), in indoor dust remains limited. In this study, indoor dust samples (n = 97) were collected from Hangzhou, China, and analyzed for PPDs and PPDQs. Results showed that nine PPDs were detected in indoor dust samples, with the total concentrations of 1.7-223 ng/g. N-(1,3-dimethylbutyl)-N'-phenyl-p-phenylenediamine (6PPD, mean 17 ng/g) was the predominant PPDs in indoor dust, followed by N, N'-di(o-tolyl)-p-phenylenediamine (DTPD, 8.6 ng/g) and N-(1,3-dimethylbutyl)-N'-(p-tolyl)-p-phenylenediamine (DMTPD, 4.7 ng/g). Five PPDQs were detected in indoor dust samples. Among detected PPDQs, 6PPDQ (14 ng/g, 0.33-82 ng/g) had the highest mean concentration, followed by DTPDQ (5.9 ng/g, < LOD-31 ng/g) and DPPDQ (2.2 ng/g, < LOD-11 ng/g). We also estimated the daily intake (DI) of PPDs and PPDQs through indoor dust ingestion. Infants had higher mean DIs of PPDs and PPDQs than children and adults. Notably, to our knowledge, this study first reports the occurrence of three novel PPDs and four novel PPDQs in indoor dust samples. More studies are needed to reveal the potential human health risks of exposure to these newly identified chemicals.

Distribution of parabens and 4-HB in human blood.

Human blood has been commonly and routinely analyzed to determine internal human exposure to parabens. However, data on the occurrence of parabens and their common metabolite, p-hydroxybenzoic acid (4-HB), in different human blood matrixes is still limited. In this study, 139 pairs of serum and whole blood samples were collected from Chinese adults, and then analyzed them for 5 parabens and 4-HB. Methylparaben (MeP) and propylparaben (PrP) were consistently the predominant parabens in human serum (mean 2.3 and 2.1 ng/mL, respectively) and whole blood (1.9 and 1.3 ng/mL, respectively). Mean concentrations of 4-HB in human serum and whole blood were 7.7 and 12 ng/mL, respectively. Concentrations of parabens, except benzylparaben (BzP), and 4-HB in human serum were significantly (p < 0.01) correlated with that in whole blood. Distribution pattern of parabens and 4-HB in human blood was evaluated, for the first time, based on their partitioning between human serum and whole blood (Kp). Mean Kp values of parabens, except BzP, increased with the alkyl chain length from 0.83 to 1.6. BzP (mean 1.4) had a comparable mean Kp value to PrP (mean 1.4). Among target analytes, 4-HB had the lowest mean Kp value (0.75). These data are important to select appropriate blood matrixes for conducting human exposure assessment and epidemiological studies on parabens.

Occurrence of p-phenylenediamine antioxidants in human urine.

General populations are widely exposed to various p-phenylenediamine antioxidants (PPDs). N-(1,3-dimethylbutyl)-N'-phenyl-p-phenylenediamine (6PPD), a typical p-phenylenediamine antioxidant, has been detected in human urine samples. However, the occurrence of other widely used PPDs in human urine is still unclear. This study comprehensively characterized the occurrence of 9 PPDs in human urine from 151 Chinese adults. Our results showed that all target PPDs were detected in human urine samples, with the total concentrations of PPDs ranging from 0.41 to 38 ng/mL. PPDs in human urine was dominated by 6PPD (mean 1.2 ng/mL, range < LOD - 3.8 ng/mL), followed by N-phenyl-N'-cyclohexyl-p-phenylenediamine (CPPD; 0.85 ng/mL,