First Evidence of Hindered Amine Light Stabilizers As Abundant, Ubiquitous, Emerging Pollutants in Dust and Air Particles: A New Concern for Human Health.

Hindered amine light stabilizers (HALSs) represent a crucial class of polymer additives that are extensively used in plastics and other polymeric materials. However, their environmental presence and related exposure risks have until now remained unexplored. This study addressed this critical knowledge by examining dust and air particles collected in South China, utilizing a comprehensive analytical approach to identify and quantify nine monomeric HALSs. A total of seven of the nine studied HALSs were detected in the samples, with bis(2,2,6,6-tetramethyl-4-piperidyl) sebacate (Tinuvin 770) and methyl 1,2,2,6,6-pentamethyl-4-piperidyl sebacate (HS-508) identified as the most abundant and widespread pollutants. Median total concentrations of HALSs ranged from 417 to 8,830 ng/g in urban dust samples and from 28.6 to 70.9 pg/m3 in urban air particles. Notably, dust concentrations of HALSs significantly exceeded those of traditional well-known light stabilizers such as UV absorbers. Human exposure assessment indicated that in contrast to air inhalation dust ingestion represented a more substantial exposure pathway owing to the relatively low volatility of these newly identified chemicals. Predictive modeling suggests that many of the examined HALSs exhibited characteristics of persistence, high toxicity, or strong potential for long-range transport, underscoring their hazardous nature. This study represents the first comprehensive investigation into the prevalence of HALSs as a class of emerging pollutants widespread in the environment, necessitating heightened attention and further research in the future.

Isolation, structure characterization and in vitro immune-enhancing activity of a glucan from the peels of stem lettuce (Lactuca sativa).

BACKGROUND: Stem lettuce is a medicinal and edible plant. The peels, accounting for 300-400 g kg-1 raw stem lettuce and containing polysaccharides 200 g kg-1 , are discarded as industrial waste, causing environment pollution and resource waste. RESULTS: A polysaccharide named PPSL10-2 was obtained from the peels of stem lettuce after hot water extraction, and gradation with cascade ultrafiltration and purification using DEAE-Sepharose cellulose. The purity and molecular weight of PPSL10-2 is 96.10% and 2.2 × 104 Da respectively, as detected by high-performance gel permeation chromatography. PPSL10-2 was found to be an α-(1→4)-d-glucan that branched at O-6 with a terminal 1-linked α-d-Glcp as side chain, and devoid of helix conformation, which was characterized by monosaccharide composition analysis, Fourier-transform infrared spectroscopy, Congo red test, scanning electron microscopy, methylation analysis and NMR spectroscopy. Furthermore, PPSL10-2 exhibited potent immune-enhancing effect by improving proliferation and phagocytosis, promoting the secretion of nitric oxide and cytokines, as well as the expression of related genes in RAW264.7 macrophages. CONCLUSION: The findings of the present study suggest that peels as an agricultural by-product of stem lettuce are good sources of polysaccharides, which could be developed as immunopotentiator for improving human health. © 2023 Society of Chemical Industry.

A review of liquid crystal monomers: Environmental occurrence, degradation, toxicity, and human exposure of an emerging class of E-waste pollutants.

Liquid crystal monomers (LCMs) are a class of organic compounds with diphenyl or dicyclohexane as the skeleton structure, which are widely used in the manufacturing of liquid crystal displays. They are recognized as novel organic compounds with persistence, bioaccumulation, toxicity, and potential for long-range transport. LCMs are inevitably released into the environment throughout the life cycle of electronic products, and their presence has been found in various abiotic matrixes (air, dust, sediment, leachate, soil) and biotic matrixes (aquatic organisms, human serum, and human skin wipe). Given that studies on LCMs are still in their infancy, this review comprehensively summarizes the extensive literature data on LCMs and identifies key knowledge gaps and future research needs. The physicochemical properties, production, and usage of LCMs are described. Their environmental distribution, degradation, toxicity, and human exposure are also discussed based on the available data and results. Existing data show that LCMs have large-scale environmental pollution and may pose potential ecological and health risks, but it is still insufficient to accurately assess their risks due to the lack of knowledge on LCMs in many areas, such as global contamination trend, environmental behavior, toxic effects, and human exposure assessment. We believe that future studies of LCMs need to investigate LCMs pollution on a large geographic scale, explore their sources, behavior, and fate in the environment, and assess their potential health hazards to organisms and humans.

Nationwide occurrence and distribution of liquid crystal monomers in municipal sewage sludge of China.

Environmental pollution and the fate of liquid crystal monomers (LCMs) in different matrices have received increasing attention owing to their potential persistence and toxicity. Sewage sludge, a representative environmental matrix, may be an important sink for LCMs. However, the contamination status of LCMs in sewage sludge remains unknown, especially on a large scale. In this study, a robust method was developed using GC-MS/MS analysis to determine 65 LCMs in sewage sludge. The occurrence of 65 LCMs in municipal sewage sludge in China was investigated for the first time. Among the 65 target LCMs, 48 were detectable, including 14 biphenyls/bicyclohexyls and their analogs (BAs) and 34 fluorobiphenyls and their analogs (FBAs). Six LCMs were detected at a rate >50 %. These results demonstrate the ubiquity of this class of synthetic chemicals in China. The total concentrations of LCMs in sludge ranged from 17.2 to 225 ng/g, with a median concentration of 46.4 ng/g. BAs were the major components of LCMs contamination in the sludge, with total BAs concentrations accounting for approximately 75 % of the total LCMs concentrations. A comparative analysis of sludge samples from different regions revealed significant regional distribution differences in LCMs: the concentrations of LCMs in sludge from East and Central China were significantly higher than those from West China (p < 0.05). Correlation and principal component analyses of the concentrations of LCMs revealed that LCMs in sludge share similar contamination sources and environmental behaviors. E-waste dismantling, domestic releases, and industrial releases may be sources of LCMs in sludge. Furthermore, the results of the degradation prediction implied that the plausible transformation products exhibited the same or even stronger persistence as the parent LCMs. Our study will be beneficial for LCMs regulation and offer suggestions for its development and safe application.

Glioma-Associated Oncogene Family Zinc Finger 2 (GLI2) Activates Wnt Signaling through Transcriptional Inhibition of Neuronal Precursor Cell-Expressed Developmentally Downregulated 4 (NEDD4L) to Promote Androgen-Induced Granulosa Cell Damage.

OBJECTIVE: Being a prevalent endocrine and metabolic disease, polycystic ovary syndrome (PCOS) severely threatens women's physical and mental health. Glioma-associated oncogene family zinc finger 2 (GLI2) expression is up-regulated in granulosa cells of PCOS patients, but its specific role in PCOS remains unclear. METHODS: Following the treatment of human ovarian granulosa cells (KGN) with dihydrotestosterone (DHT), RT-qPCR and western blot were utilized to check GLI2 expression. After GLI2 expression was silenced, cell activity was detected through CCK8 and apoptosis was examined via TUNEL and western blot. Inflammation and oxidative stress were tested utilizing ELISA and western blot. The binding between GLI2 and neuronal precursor cell-expressed developmentally downregulated 4 (NEDD4L) promoter was predicted by JASPAR database and verified by luciferase reporter and ChIP assay. In addition, RT-qPCR and western blot were applied to check the mRNA and protein expressions of NEDD4L. Following the knockdown of NEDD4L in GLI2-silencing cells, CCK8 assay, TUNEL assay, western blot, ELISA and other methods were performed again. Finally, western blot detected the expressions of Wnt pathway-related proteins. RESULTS: GLI2 was up-regulated in DHT-treated KGN cells. Interference with GLI2 increased the viability, decreased the apoptosis, and inhibited the inflammatory response and oxidative stress of DHT-induced KGN cells. GLI2 could bind to NEDD4L promoter and transcriptionally suppress NEDD4L expression. Further experiments testified that NEDD4L depletion reversed the impacts of GLI2 deficiency on the viability, apoptosis, inflammation, oxidative stress and Wnt signaling pathway in DHT-challenged KGN cells. CONCLUSION: GLI2 activated Wnt signaling to promote androgen-induced granulosa cell damage through transcriptional inhibition of NEDD4L.

Widespread Occurrence and Transport of p-Phenylenediamines and Their Quinones in Sediments across Urban Rivers, Estuaries, Coasts, and Deep-Sea Regions.

p-Phenylenediamines (PPDs) are widely used as antioxidants in tire rubber, and their derived quinone transformation products (PPD-Qs) may pose a threat to marine ecosystems. A compelling example is N-(1,3-dimethylbutyl)-N'-phenyl-p-phenylenediamine (6PPD)-derived quinone, called 6PPD-Q, as the causal toxicant for stormwater-linked acute mortality toward coho salmon. However, the knowledge of the co-occurrences of PPDs and PPD-Qs and their transport from freshwater to oceanic waterbodies on a large geographical scale remains unknown. Herein, we performed the first large-scale survey of these chemicals in sediments across urban rivers, estuaries, coasts, and deep-sea regions. Our results demonstrated that seven PPDs and four PPD-Qs are ubiquitously present in riverine, estuarine, and coastal sediments, and most of them also occur in deep-sea sediments. The most dominant chemicals of concern were identified as 6PPD and 6PPD-Q. Total sedimentary concentrations of PPDs and PPD-Qs presented a clear spatial trend with decreasing levels from urban rivers (medians: 39.7 and 15.2 ng/g) to estuaries (14.0 and 5.85 ng/g) and then toward coasts (9.47 and 2.97 ng/g) and deep-sea regions (5.24 and 3.96 ng/g). Interestingly, spatial variation in the ratios of 6PPD to 6PPD-Q (R6PPD/6PPD-Q) also presented a clear decreasing trend. Our field measurements implied that riverine outflows of PPDs and PPD-Qs may be an important route to transport these tire rubber-derived chemicals to coastal and open oceans.

Clinical Research into Treating Unexplained Recurrent Spontaneous Abortion during Early Pregnancy with the Qing Yi Tiao Mian Formula.

OBJECTIVE: The present study aims to analyze the clinical effect of the Qing Yi Tiao Mian (QYTM) formula on unexplained recurrent spontaneous abortion (URSA) during early pregnancy and the immune balance of T lymphocytes. METHODS: With their consent, 45 patients with URSA in weeks 4-9 of pregnancy were separated into three groups, i.e., the conventional fetal protection (n = 15), prednisone treatment (n = 10), and QYTM formula treatment (n = 20) groups. These patients received treatment once they had been diagnosed with an intrauterine pregnancy. The conventional fetal protection group was given progesterone (20 ∼ 40 mg daily injection) for four weeks. The prednisone treatment group was given progesterone (20 ∼ 40 mg daily injection) + prednisone (5 mg/d) for four weeks. The QYTM formula treatment group was given progesterone (20 ∼ 40 mg daily injection) + QYTM formula (one dose per day) for four weeks. In addition, women who had previously had a normal pregnancy were enrolled as a control group (n = 18). The success rate of the pregnancy in the first trimester was observed in each group, and the proportion of T lymphocytes in the peripheral blood before and after treatment was recorded. RESULTS: Among the 20 patients with URSA in the QYTM formula treatment group, 19 remained pregnant. Thus, the success rate during early pregnancy was 95%, which was significantly higher than the conventional fetal protection (53.33%) and prednisone treatment (70%) groups. The CD8+ T and natural killer (NK) cells population in the URSA groups was higher compared with the control group (P < 0.01). The QYTM formula treatment significantly decreased the ratio of CD8+ T lymphocytes (P < 0.01) and NK cells (P < 0.01). CONCLUSION: The QYTM formula significantly decreased the spontaneous abortion rate in patients with URSA during early pregnancy. The mechanism may be closely related to the inhibition of the killer lymphocytes' proliferation by CD8+ T lymphocytes and NK cells.

Molecular structure on the detoxification of fluorinated liquid crystal monomers with reactive oxidation species in the photocatalytic process.

Fluorinated liquid crystal monomers (LCMs) are begun to emerge as new persistent organic pollutants. Herein, the structure-reactivity relationships of fluorinated LCMs 1,2,3-trifluoro-5-[3-(3-propylcyclohexyl)cyclohexyl]benzene (TPrCB), 1,2-difluoro-4-[trans-4-(trans-4-propylcyclohexyl)cyclohexyl]benzene (DPrCB), 4-[(trans,trans)-4'-(3-Buten-1-yl)[1,1'-bicyclohexyl]-4-yl]-1,2-difluoro-benzene (BBDB) and 1-[4-(4-ethylcyclohexyl)cyclohexyl]-4(trifluoromethoxy)benzene (ECTB) subject to photocatalysis-generated oxidation species were investigated. The degradation rate constant of BBDB was 3.0, 2.6, and 6.8 times higher than DPrCB, TPrCB and ECTB, respectively. The results reveal that BBDB, DPrCB and TPrCB had mainly negative electrostatic potential (ESP) regions which were vulnerable to electrophilic attack by h+, •OH and •O2 -, while ECTB was composed of mainly positive ESP regions which were vulnerable to nucleophilic attack by •OH and •O2 -. The detoxification processes of BBDB, DPrCB and TPrCB included carbon bond cleavage and benzene ring opening. However, the methoxy group of ECTB reduced the nucleophilic reactivity on the benzene ring, leading to slower detoxification efficiency. These findings may help to develop LCMs treatment technologies based on structure-reactivity relationships.

Identification of Fluorescent Brighteners as Another Emerging Class of Abundant, Ubiquitous Pollutants in the Indoor Environment.

Fluorescent brighteners (FBs) are extensively used as important chemical additives in multiple industrial fields worldwide. The history of the use of global FBs spans over 60 years, but knowledge on their environmental occurrence and risks remains largely unknown. Here, we screened indoor dust and hand wipes from South China for a broad suite of 17 emerging FBs using a new comprehensive analytical method. All 17 FBs were detected in the indoor environment for the first time, most of them having been rarely investigated or never reported in prior environmental studies. Ionic FBs were found to be more abundant than nonionic ones. The median total concentrations of the 17 detectable FBs reached 11,000 ng/g in indoor dust and 2640 ng/m2 in hand wipes, comparable to or higher than those of well-known indoor pollutants. Human exposure assessment indicated that hand-to-mouth contact is a significant pathway for exposure to FBs, with a comparable contribution to that of dust ingestion. Most of the newly identified FBs are predicted to have persistent, bioaccumulative, or toxic properties. Our work demonstrates that FBs are another class of highly abundant, hazardous, and ubiquitous indoor pollutants that have been overlooked for decades and points to an emerging concern.

Release and Gas-Particle Partitioning Behavior of Liquid Crystal Monomers during the Dismantling of Waste Liquid Crystal Display Panels in E-Waste Recycling Facilities.

Liquid crystal monomers (LCMs) are a class of emerging chemical pollutants; however, their release and gas-particle partitioning remain unknown. This study performed the first comprehensive analysis of a wide range of 93 LCMs in the ambient air of liquid crystal display (LCD) dismantling facilities. A total of 53 of the 93 target LCMs were detected in the air samples. The total atmospheric concentrations (gas and particles) of LCMs (∑LCMs) ranged from 68,800 to 385,000 (median of 204,000) pg/m3. Most LCMs were predominant in the gas phase, implying that their atmospheric transport would be mainly governed by gas rather than particle diffusions. Differential distribution patterns of the LCMs were observed due to their different atmospheric partitioning behaviors. Significant linear correlations were found between the gas-particle partitioning coefficients (KP) and the predicted subcooled vapor pressures (PL) and octanol-air partitioning coefficients (Koa) (p < 0.01). Compared with two equilibrium-state models, the experimentally observed particulate fractions (ϕ) fit better with the predicted values based on the Li-Ma-Yang (L-M-Y) steady-state model, and Koa was identified as a key factor determining the atmospheric fate pathways of LCMs. Our study highlights another new class of chemicals significantly contributing to the chemical mixture in the ambient air at e-waste recycling areas.

Trace analysis of multiple synthetic phenolic antioxidants in foods by liquid chromatography-tandem mass spectrometry with complementary use of electrospray ionization and atmospheric pressure chemical ionization.

This study presented a universal LC-MS/MS method for trace analysis of multiple synthetic phenolic antioxidants (SPAs) in foods by complementary use of electrospray ionization (ESI) and atmospheric pressure chemical ionization (APCI). The analytes included not only the well-known BHT and BHA but also 18 high molecular weight SPAs. The method utilized APCI to achieve sensitive analysis of BHT, Irganox 1010, Irganox 330, and Irganox 3125 based on the finding that APCI significantly improved the sensitivity of these weakly acidic or slightly polar SPAs, and utilized ESI to obtain sensitive analysis of other SPAs. Additionally, the method avoided background contamination by using effective measures including installation of a trapping column in the LC system. Method performance assessment showed satisfactory sensitivity, linearity, accuracy, and precision for analysis of SPAs in vegetable oil, milk powder, and baby fruit puree. Method application revealed widespread contamination of foods with BHT, Irganox 1010, and Irganox 1076.

Comprehensive Identification of Liquid Crystal Monomers-Biphenyls, Cyanobiphenyls, Fluorinated Biphenyls, and their Analogues-in Waste LCD Panels and the First Estimate of their Global Release into the Environment.

Our previous study demonstrated massive emissions of liquid crystal monomers (LCMs) from liquid crystal display (LCD)-associated e-waste dismantling; however, the compositions, priority list, and inventory of LCMs in waste LCD panels remain unknown. Herein, we conducted the first comprehensive identification covering a broader range of LCMs, including 21 biphenyls and analogues (BAs), 28 cyanobiphenyls and analogues (CBAs), and 44 fluorinated biphenyls and analogues (FBAs), in waste television/computer LCD panels. A total of 64 of the 93 target LCMs, including 19 BAs, 6 CBAs, and 39 FBAs, were widely detected in collected waste LCD panels. Approximately 10-18 of the 64 detectable LCMs were identified as the main compositions in various waste LCD panels, which contributed to >90% of the total LCMs. Total concentrations of FBAs in the television/computer LCD panel samples were comparable to those of BAs but much higher than those of CBAs, indicating FBAs and BAs being the commonly used LCM categories. The composition distribution of LCMs varied between television/computer LCDs and among different brands of television/computer LCDs. A preliminary estimate of the globally direct release of LCMs from waste television/computer LCD panels into various environmental compartments was about 1.07-107 kg/year, which will increase considerably in the near future.

Associations of Prenatal Exposure to Per- and Polyfluoroalkyl Substances with the Neonatal Birth Size and Hormones in the Growth Hormone/Insulin-Like Growth Factor Axis.

Toxicological data suggest a significant developmental toxicity of per- and polyfluoroalkyl substances (PFASs); however, evidence in humans remains inconclusive. Furthermore, the effects of prenatal exposure to PFASs on hormones in the growth hormone (GH)/insulin-like growth factor (IGF) axis of newborns remain largely unclear. We aimed to investigate the associations of prenatal exposure to PFASs with the neonatal birth size, GH, IGF-1, and IGF-binding protein 3 (IGFBP-3). The concentrations of 22 PFASs were measured in the plasma of 224 pregnant women collected within 3 days before delivery (39.3 weeks) in Guangzhou, China, and the anthropometric data were gathered from medical records. Paired cord blood was collected at delivery to determine GH, IGF-1, and IGFBP-3 levels. Multivariable linear regression models revealed the inverse associations of several long-chain PFASs with birth weight and ponderal index as well as the significant associations of perfluorobutanoic acid and perfluorooctanoic acid (PFOA) with IGFBP-3 levels. The Bayesian kernel machine regression confirmed the association of perfluorooctane sulfonate with birth weight and ponderal index and of PFOA with IGFBP-3 and identified an inverse joint effect of exposure to a mixture of multiple PFASs on birth weight. The findings provide the first comprehensive evidence on the individual and joint effects of multiple PFASs on the neonatal birth size and hormones in the GH/IGF axis, which requires further confirmation.

Hair and nails as noninvasive bioindicators of human exposure to chlorinated paraffins: Contamination patterns and potential influencing factors.

Most of the studies on short- and medium-chain chlorinated paraffins (SCCPs and MCCPs) in human tissues have focused on human milk and blood. However, little is known about the occurrence of CPs in human hair and nails. In this study, SCCPs and MCCPs were analyzed in 62 pairs of human hair and nails from North China. Median concentrations (range) of SCCPs and MCCPs in human hair were 239 (19.2-877) and 325 (16.9-893) ng/g dw, respectively, all of which were significantly higher than 154 (57.7-355) and 233 (61.0-476) ng/g dw, respectively, in nail samples (p < 0.05). The homologue profiles of CPs in human hair were similar to those in nails, where SCCPs and MCCPs were dominated by C10Cl6-7 and C14Cl7-8, respectively. A significant positive relationship was observed between CP levels and age of people for hair, whereas negative linear correlations were observed for nails. The redundancy analysis indicated that age of people might be the main influencing factor on the accumulation of CPs in hair and nails. The present study performed comprehensive evaluation of CP exposure levels in human hair and nail and highlights the need for more data on relationship between internal and external exposure to CPs.

Identification of Environmental Liquid-Crystal Monomers: A Class of New Persistent Organic Pollutants-Fluorinated Biphenyls and Analogues-Emitted from E-Waste Dismantling.

Liquid-crystal monomers (LCMs), especially fluorinated biphenyls and analogues (FBAs), are considered to be a new generation of persistent, bioaccumulative, and toxic organic pollutants, but their emissions from liquid-crystal display (LCD)-associated e-waste dismantling remain unknown. To fill this knowledge gap, a broad range of 46 LCMs, including 39 FBAs and 7 biphenyls/bicyclohexyls and analogues (BAs), were investigated by a dedicated target analysis in e-waste dust samples. Of 39 target FBAs, 34 were detected in LCD dismantling-associated dust. Among these 34 detectable FBAs, 9 were detected in 100% of the samples and 25 were frequently detected in >50% of the samples. The total concentrations of these 34 FBAs (∑34FBAs) detected in LCD e-waste dust were in the range of 225-976,000 (median: 18,500) ng/g, significantly higher than those in non-LCD e-waste dust (range: 292-18,500, median: 2300 ng/g). In addition to FBAs, six of seven BAs were also frequently detected in LCD e-waste dust with total concentrations (∑6BAs) of 29.8-269,000 (median: 3470) ng/g. Very strong and significant correlations (P < 0.01) were identified in all frequently detected LCMs, indicating their common applications and similar sources. Our findings demonstrate that e-waste dismantling contributes elevated emissions of FBAs and BAs to the ambient environment.

A Type VI Secretion System Facilitates Fitness, Homeostasis, and Competitive Advantages for Environmental Adaptability and Efficient Nicotine Biodegradation.

Gram-negative bacteria employ secretion systems to translocate proteinaceous effectors from the cytoplasm to the extracellular milieu, thus interacting with the surrounding environment or microniche. It is known that bacteria can benefit from the type VI secretion system (T6SS) by transporting ions to combat reactive oxygen species (ROS). Here, we report that T6SS activities conferred tolerance to nicotine-induced oxidative stress in Pseudomonas sp. strain JY-Q, a highly active nicotine degradation strain isolated from tobacco waste extract. AA098_13375 was identified to encode a dual-functional effector with antimicrobial and anti-ROS activities. Wild-type strain JY-Q grew better than the AA098_13375 deletion mutant in nicotine-containing medium by antagonizing increased intracellular ROS levels. It was, therefore, tentatively designated TseN (type VI secretion system effector for nicotine tolerance), homologs of which were observed to be broadly ubiquitous in Pseudomonas species. TseN was identified as a Tse6-like bacteriostatic toxin via monitoring intracellular NAD+ TseN presented potential antagonism against ROS to fine tune the heavy traffic of nicotine metabolism in strain JY-Q. It is feasible that the dynamic tuning of NAD+ driven by TseN could satisfy demands from nicotine degradation with less cytotoxicity. In this scenario, T6SS involves a fascinating accommodation cascade that prompts constitutive biotransformation of N-heterocyclic aromatics by improving bacterial robustness/growth. In summary, the T6SS in JY-Q mediated resistance to oxidative stress and promoted bacterial fitness via a contact-independent growth competitive advantage, in addition to the well-studied T6SS-dependent antimicrobial activities.IMPORTANCE Mixtures of various pollutants and the coexistence of numerous species of organisms are usually found in adverse environments. Concerning biodegradation of nitrogen-heterocyclic contaminants, the scientific community has commonly focused on screening functional enzymes that transform pollutants into intermediates of attenuated toxicity or for primary metabolism. Here, we identified dual roles of the T6SS effector TseN in Pseudomonas sp. strain JY-Q, which is capable of degrading nicotine. The T6SS in strain JY-Q is able to deliver TseN to kill competitors and provide a growth advantage by a contact-independent pattern. TseN could monitor the intracellular NAD+ level by its hydrolase activity, causing cytotoxicity in competitive rivals but metabolic homeostasis on JY-Q. Moreover, JY-Q could be protected from TseN toxicity by the immunity protein TsiN. In conclusion, we found that TseN with cytotoxicity to bacterial competitors facilitated the nicotine tolerance of JY-Q. We therefore reveal a working model between T6SS and nicotine metabolism. This finding indicates that multiple diversified weapons have been evolved by bacteria for their growth and robustness.

Additional Role of Nicotinic Acid Hydroxylase for the Transformation of 3-Succinoyl-Pyridine by Pseudomonas sp. Strain JY-Q.

Nicotine and nicotinic acid (NA) are both considered to be representatives of N-heterocyclic aromatic compounds, and their degradation pathways have been revealed in Pseudomonas species. However, the cooccurrence of these two pathways has only been observed in Pseudomonas sp. strain JY-Q. The nicotine pyrrolidine catabolism pathway of strain JY-Q consists of the functional modules Nic1, Spm, and Nic2. The module enzyme, 3-succinoylpyridine monooxygenase (Spm), catalyzes transformation of 3-succinoyl-pyridine (SP) to 6-hydroxy-3-succinoyl-pyridine (HSP). There exist two homologous but not identical Spm enzymes (namely, Spm1 and Spm2) in JY-Q. However, when spm1 and spm2 were both in-frame deleted, the mutant still grew well in basic salt medium (BSM) supplemented with nicotine as the sole carbon/nitrogen nutrition, suggesting that there exists an alternative pathway responsible for SP catabolism in JY-Q. NicAB, an enzyme accounting for NA hydroxylation, contains reorganized domains similar to those of Spm. When the JY-Q_nicAB gene (nicAB in strain JY-Q) was introduced into another Pseudomonas strain, one that is unable to degrade NA, the resultant recombinant strain exhibited the ability to transform SP to HSP, but without the ability to metabolize NA. Here, we conclude that NicAB in strain JY-Q exhibits an additional role in SP transformation. The other genes in the NA cluster, NicXDFE (Nic2 homolog), then also exhibit a role in subsequent HSP metabolism for energy yield. This finding also suggests that the cooccurrence of nicotine and NA degradation genes in strain JY-Q represents an advantage for JY-Q, making it more effective and flexible for the degradation of nicotine.IMPORTANCE 3-Succinoyl-pyridine (SP) and 6-hydroxy-3-succinoyl-pyridine (HSP) are both valuable chemical precursors to produce insecticides and hypotensive agents. SP and HSP could be renewable through the nicotine microbial degradation pathway, in which 3-succinoylpyridine monooxygenases (Spm) account for transforming SP into HSP in Pseudomonas sp. strain JY-Q. However, when two homologous Spm genes (spm1 and spm2) were knocked out, the mutant retained the ability to degrade nicotine. Thus, in addition to Spm, JY-Q should have an alternative pathway for SP conversion. In this research, we showed that JY-Q_NicAB was responsible for this alternative SP conversion. Both of the primary functions for nicotinic acid dehydrogenation and the additional function for SP metabolism were detected in a recombinant strain harboring JY-Q_NicAB. As a result, both nicotinic acid and nicotine degradation pathways in JY-Q contribute to its remarkable nicotine tolerance and nicotine degradation availability. These findings also provide one more metabolic engineering strategy for accumulation for value-added intermediates.

Expression and functional identification of two homologous nicotine dehydrogenases, NicA2 and Nox, from Pseudomonas sp. JY-Q.

Nicotine contamination in tobacco waste effluent (TWE) from tobacco industry is a serious threat to public health and environment. Microbial degradation is an impending approach to remove nicotine and transform it into some other high value chemicals. Pseudomonas sp. JY-Q exhibits high efficiency of degradation, which can degrade 5 g/L of nicotine within 24 h. In strain JY-Q, we found the co-occurrence of two homologous key enzymes NicA2 and Nox, which catalyze nicotine to N-methylmyosmine, and then to pseudooxylnicotine via simultaneous hydrolysis. In this study, recombinant NicA2 and Nox were expressed in E. coli BL21(DE3) and purified. In vitro, the activity of recombinant NicA2 and Nox was accelerated by adding co-factor NAD+, suggesting that they worked as dehydrogenases. The optimal reaction conditions, substrate affinity, catabolism efficiency, pH-stability and thermal-stability were determined. Nox showed lower efficiency, but at a higher stability level than NicA2. Nox exhibited wider pH range and higher temperature as optimal conditions for the enzymatic reaction. In addition, The Nox showed higher thermo-stability and acid-stability than that of NicA2. The study on enzymatic reaction kinetics showed that Nox had a lower Km and higher substrate affinity than NicA2. These results suggest that Nox plays more significant role than NicA2 in nicotine degradation in TWE, which usually is processed at low pH (4-5) and high temperature (above 40 °C). Genetic engineering is required to enhance the affinity and suitability of NicA2 for an increased additive effect on homologous NicA2 and Nox in strain JY-Q.

Beyond Traditional Organophosphate Triesters: Prevalence of Emerging Organophosphate Triesters and Organophosphate Diesters in Indoor Dust from a Mega E-waste Recycling Industrial Park in South China.

Numerous studies have reported the environmental contamination with traditional organophosphate triesters (tri-OPEs), but there is very little information on emerging tri-OPEs and organophosphate diesters (di-OPEs), especially in e-waste recycling areas. In this study, we conducted a comprehensive survey to monitor a broad suite of 11 traditional tri-OPEs, 12 emerging OPEs, and 10 di-OPEs in indoor dust collected from the workshops of (n = 42) and residential homes adjacent to (n = 24) a mega e-waste recycling industrial park in South China. In addition to traditional tri-OPEs, all of the emerging OPEs and di-OPEs were frequently detected in the dust samples. Total concentrations of emerging tri-OPEs and di-OPEs were in the range of 1210-62 900 and 2010-55 600 ng/g in the workshop dust and 435-23 700 and 186-4350 ng/g in the local home dust, respectively, which were comparable to those of traditional tri-OPEs (1160-61 500 and 370-13 900 ng/g, respectively). Most OPEs exhibited significantly higher concentrations in workshop dust versus local home dust (p < 0.05), indicating that e-waste dismantling activities contributed to the high residues of OPEs in indoor dust. Correlation analysis revealed that tri-OPEs have some common emission sources, i.e., e-waste and household products, while di-OPEs could originate from different sources, e.g., tri-OPE degradation, direct commercial application, and impurities in tri-OPE formulas. For both occupational workers and local adults, the median estimated daily intake values of emerging tri-OPEs (7.5 and 1.7 ng/kg bw/day, respectively) and di-OPEs (3.9 and 0.2 ng/kg bw/day, respectively) were comparable to that of traditional tri-OPEs (4.3 and 1.0 ng/kg bw/day, respectively), which suggests the important contribution of the emerging tri-OPEs and di-OPEs to the overall risks of human external exposure to OPE chemicals.