Current research hotspots and frontier trends on carbon budget of coastal wetlands: A bibliometric analysis.

Coastal wetlands are the main distribution of blue carbon in coastal zones and well known for their high carbon sequestration capacity. Investigating the variation of carbon budget is crucial for understanding the functionality of coastal wetlands and effectively addressing climate change. In this study, a bibliometric analysis of 4,509 articles was conducted to reveal research progress, hot issues, and emerging trends in the coastal wetland carbon budget field. The number of publications and citations in this field increased exponentially from 1991 to 2022. The leading subject category was Environmental Sciences with 1,844 articles (40.9%). At present, studies have been focused on blue carbon, the effects of climate change and man-made disturbances on carbon cycle, and the restoration of coastal wetlands. Based on the hotspots and trends in this field, the future researches should include (1) exploring the functional mechanisms of various factors affecting carbon cycle and establishing a methodological system for the estimation of blue carbon in coastal wetlands; (2) researching restoration techniques of coastal wetland and constructing wetland restoration evaluation index system; and (3) formulating enforceable carbon trading policy and strengthening international cooperation.

Comparison study on enhancement of phosphorus recovery from low-strength wastewater treated with different magnesium-based electrochemical constructed wetland.

Phosphorus (P) recovery from wastewaters treated with constructed wetlands (CWs) could alleviate the current global P crisis but has not received sufficient attention. In this study, P transformation in different magnesium-based electrochemical CWs, including micro-electrolysis CW (M-CW), primary battery CW (P-CW), and electrolysis CW (E-CW), was thoroughly examined. The results revealed that the P removal efficiency was 53.0%, 75.8%, and 61.9% in the M-CW, E-CW, and P-CW, respectively. P mass balance analysis showed that P electrode deposition was the main reason for the higher P removal in the E-CW and P-CW. Significant differences were found between the E-CW and P-CW, P was distributed primarily on the magnesium plate in the P-CW but was distributed on the carbon plate in the E-CW. The E-CW had excellent P recovery capacity, and struvite was the major P recovery product. More intense magnesium plate corrosion and alkaline environment increased struvite precipitation in the E-CW, with the proportion of 61.6%. The results of functional microbial community analysis revealed that the abundance of electroactive bacteria was positively correlated with the deposition of struvite. This study provided an essential reference for the targeted electrochemical regulation of electric field processes and microorganisms in CWs to enhance P recovery.

Influence of iron species on the simultaneous nitrate and sulfate removal in constructed wetlands under low/high COD concentrations.

Nitrate and sulfate are crucial factors of eutrophication and black and odorous water in the surface water and thus have raised increasing environmental concerns. Constructed wetlands (CWs) are the last ecological barrier before effluent enters the natural water body. To explore the simultaneous removal of nitrate and sulfate, the CW microcosms of CW-Con (with quartz sand), CW-ZVI (quartz sand and zero-valent iron), CW-Mag (quartz sand and magnetite), CW-ZVI + Mag (quartz sand, ZVI and magnetite) groups were set up under the low (100 mg/L)/high (300 mg/L) chemical oxygen demand (COD) concentration. Under the high COD condition, CW-ZVI group showed the best performance in nitrate (97.1%) and sulfate (96.9%) removal. Under the low COD concentration, the removal content of nitrate and sulfate in CW-ZVI group was better than CW-Mag group. The reason for this result was that zero-valent iron (ZVI) could be the electron donor for nitrate and sulfate reduction. Meanwhile, ZVI promoted chemical denitrification under high COD concentration according to PCA analysis. In addition, the produced sulfides inhibited the relative abundance of denitrifying bacteria, resulting in the lowest nitrate removal rate in CW-Mag group with sufficient electron donors. This study provided an alternative method to enhance simultaneous sulfate and nitrate removal in CWs.

Comprehensive evaluation of manganese oxides and iron oxides as metal substrate materials for constructed wetlands from the perspective of water quality and greenhouse effect.

Manganese oxides and iron oxides have been widely introduced in constructed wetlands (CWs) for sewage treatment due to their extensiveness in nature and their ability to participate in various reactions, but their effects on greenhouse gas (GHG) emissions remain unclear. Here, a set of vertical subsurface-flow CWs (Control, Fe-VSSCWs, and Mn-VSSCWs) was established to comprehensively evaluate which are the better metal substrate materials for CWs, iron oxides or manganese oxides, through water quality and the global warming potential (GWP) of nitrous oxide (N2O), methane (CH4), and carbon dioxide (CO2). The results revealed that the removal efficiencies of chemical oxygen demand (COD), total nitrogen (TN), and total phosphorus (TP) in Mn-VSSCWs were all higher than that in Fe-VSSCWs, and manganese oxides could almost completely suppress the CH4 production and reduce GWP (from 8.15 CO2-eq/m2/h to 7.17 mg CO2-eq/m2/h), however, iron oxides promoted GWP (from 8.15 CO2-eq/m2/h to 10.84 mg CO2-eq/m2/h), so manganese oxides are the better CW substrate materials to achieve effective sewage treatment while reducing the greenhouse gas effect.

Intensive removal of PAHs in constructed wetland filled with copper biochar.

In this study, biochar-loading copper ions (Cu-BC), a novel composite for removing phenanthrene very efficiently from water, was prepared using the impregnation method. The performance of constructed wetlands (CWs) with these modified and original biochar as substrates was analyzed. CW with Cu-BC removed a large amount of phenanthrene (94.09 ± 3.02%). According to the surface characteristics analysis, Cu-BC can promote the removal of pollutants via complex absorption, hydrophobic adsorption, increasing the Lewis Pair and electrostatic attraction. Furthermore the higher nitrate removal rate in the treated system (91.11 ± 1.17%) was observed to have higher levels of bacterial metabolic diversity and denitrifier types. The phenanthrene accumulated in plants with this treatment system was enhanced by the role of copper in photosynthesis. It is able to boost the plant extraction of organic matter.

6-Hydroxypseudooxynicotine Dehydrogenase Delivers Electrons to Electron Transfer Flavoprotein during Nicotine Degradation by Agrobacterium tumefaciens S33.

Agrobacterium tumefaciens S33 degrades nicotine via a novel hybrid of the pyridine and the pyrrolidine pathways. The hybrid pathway consists of at least six steps involved in oxidoreductive reactions before the N-heterocycle can be broken down. Collectively, the six steps allow electron transfer from nicotine and its intermediates to the final acceptor O2 via the electron transport chain (ETC). 6-Hydroxypseudooxynicotine oxidase, renamed 6-hydroxypseudooxynicotine dehydrogenase in this study, has been characterized as catalyzing the fourth step using the artificial electron acceptor 2,6-dichlorophenolindophenol. Here, we used biochemical, genetic, and liquid chromatography-mass spectrometry (LC-MS) analyses to determine that 6-hydroxypseudooxynicotine dehydrogenase utilizes the electron transfer flavoprotein (EtfAB) as the physiological electron acceptor to catalyze the dehydrogenation of pseudooxynicotine, an analogue of the true substrate 6-hydroxypseudooxynicotine, in vivo, into 3-succinoyl-semialdehyde-pyridine. NAD(P)+, O2, and ferredoxin could not function as electron acceptors. The oxygen atom in the aldehyde group of the product 3-succinoyl-semialdehyde-pyridine was verified to be derived from H2O. Disruption of the etfAB genes in the nicotine-degrading gene cluster decreased the growth rate of A. tumefaciens S33 on nicotine but not on 6-hydroxy-3-succinoylpyridine, an intermediate downstream of the hybrid pathway, indicating the requirement of EtfAB for efficient nicotine degradation. The electrons were found to be further transferred from the reduced EtfAB to coenzyme Q by the catalysis of electron transfer flavoprotein:ubiquinone oxidoreductase. These results aid in an in-depth understanding of the electron transfer process and energy metabolism involved in the nicotine oxidation and provide novel insights into nicotine catabolism in bacteria.IMPORTANCE Nicotine has been studied as a model for toxic N-heterocyclic aromatic compounds. Microorganisms can catabolize nicotine via various pathways and conserve energy from its oxidation. Although several oxidoreductases have been characterized to participate in nicotine degradation, the electron transfer involved in these processes is poorly understood. In this study, we found that 6-hydroxypseudooxynicotine dehydrogenase, a key enzyme in the hybrid pyridine and pyrrolidine pathway for nicotine degradation in Agrobacterium tumefaciens S33, utilizes EtfAB as a physiological electron acceptor. Catalyzed by the membrane-associated electron transfer flavoprotein:ubiquinone oxidoreductase, the electrons are transferred from the reduced EtfAB to coenzyme Q, which then could enter into the classic ETC. Thus, the route for electron transport from the substrate to O2 could be constructed, by which ATP can be further sythesized via chemiosmosis to support the baterial growth. These findings provide new knowledge regarding the catabolism of N-heterocyclic aromatic compounds in microorganisms.

Emissions of fine particulate nitrated phenols from various on-road vehicles in China.

Nitrated phenols are receiving increasing attention due to their adverse impacts on the environment and human health. Previous measurements have revealed the non-ignorable contribution of vehicle exhaust to atmospheric nitrated phenols in urban areas. However, there is a lack of comprehensive understanding of the emission characteristics and the total emission of nitrated phenols from current on-road traffic. This study investigated the emissions from eight passenger vehicles, eight trucks, and two taxis, with fuel types including diesel, gasoline, and compressed natural gas. Exhaust emissions were collected and measured using a mobile measurement system on two testing routes. Twelve nitrated phenols in the collected fine particulate matter were detected using ultrahigh performance liquid chromatography-mass spectrometry. Overall, the emission profiles of fine particulate nitrated phenols varied with vehicle load and fuel type. The 4-nitrophenol and its methyl derivatives were dominant nitrated phenol species emitted by the vehicles with proportions of 38.4%-68.0%, which is significantly different from the proportions of nitrated phenols emitted from biomass burning and coal combustion. The emission factors also exhibited large variations across vehicle type, fuel type, and emission standards, with relatively low values for gasoline vehicles and taxis fueled by compressed natural gas and high values for diesel vehicles. Based on the emission factors of nitrated phenols from different vehicles, the estimated total emission of nitrated phenols from on-road vehicles in China was 58.9 Mg (-86%-85% within 95% confidence interval), with diesel trucks contributing the most substantial fractions. This work highlights the very high level of emissions of nitrated phenols from diesel vehicles and provides an essential basis for atmospheric modeling and effective pollution control.

Influence of application of manganese ore in constructed wetlands on the mechanisms and improvement of nitrogen and phosphorus removal.

Vertical up-flow constructed wetlands (CWs) with manganese ore (Mn ore) as media (M-CWs) were developed to treat simulated polluted river water. The results showed that the average removal efficiencies for NH4-N, NO3-N, TN and TP were 91.74%, 83.29%, 87.47% and 65.12% in M-CWs, respectively, which were only 79.12%, 72.90%, 75.85% and 43.23% in the CWs without Mn ore (C-CWs). Nutrient mass balance showed that nitrogen (N) removal was improved by enhanced microbial processes, media storage and plant uptake in M-CWs. Moreover, almost 50% of phosphorus (P) was retained by media storage because of the adsorption processes on Mn ore. It was found that addition of Mn ore enhanced denitrification as the relative abundance of denitrifying bacteria increased. The produced Mn(II) and more abundant Gammaproteobacteria confirmed alternative N removal pathways including anoxic nitrification coupled to Mn ore reduction and denitrification using Mn(II) as electron donor. Mn(II) concentration in the effluent of M-CWs was below the drinking water limit of 0.1 mg/L, which makes them environmentally-friendly.

Fabrication of pH-Sensitive Tetramycin Releasing Gel and Its Antibacterial Bioactivity against Ralstonia solanacearum.

Ralstonia solanacearum (R. solanacearum)-induced bacterial wilt of the nightshade family causes a great loss in agricultural production annually. Although there has been some efficient pesticides against R. solanacearum, inaccurate pesticide releasing according to the onset time of bacterial wilt during the use of pesticides still hinders the disease management efficiency. Herein, on the basis of the soil pH change during R. solanacearum growth, and pH sensitivity of the Schiff base structure, a pH-sensitive oxidized alginate-based double-crosslinked gel was fabricated as a pesticide carrier. The gel was prepared by crosslinking oxidized sodium alginate (OSA) via adipic dihydrazide (ADH) and Ca2+. After loading tetramycin into the gel, it showed a pH-dependent pesticide releasing behavior and anti-bacterial activity against R. solanacearum. Further study also showed that the inhibition rate of the tetramycin-loaded gel was higher than that of industrial pesticide difenoconazole. This work aimed to reduce the difficulty of pesticide administration in the high incidence period of bacterial wilt and we believe it has a great application potential in nightshade production.

Microbial nitrogen removal of ammonia wastewater in poly (butylenes succinate)-based constructed wetland: effect of dissolved oxygen.

Constructed wetland (CW) is popular in wastewater treatment for its prominent advantage of low construction and operation cost. However, the nitrogen removal in conventional CW is usually limited by the low dissolved oxygen (DO) and insufficient electron donor. This paper investigated the nitrogen removal performance and mechanisms in the poly (butylenes succinate)-based CW (PBS-CW) while treating ammonia wastewater under different DO levels. The average DO contents in limited-aeration and full-aeration phases were 1.68 mg L-1 and 5.71 mg L-1, respectively. Results indicated that, with the ammonia nitrogen loading rate of 25 g N m-3 day-1, total nitrogen removal ratios in the PBS-CW under the limited-aeration and full-aeration phases were 72% and 99%, respectively. Combined analyses revealed that simultaneous nitrification and denitrification (SND) via nitrite/nitrate were the main microbial nitrogen removal pathways in the aeration phase of the PBS-CW (> 89%). The microbial carrier of biodegradable material was believed to play a significant role in prompting SND performance while dealing with low C/N wastewater. Due to the coexistence of micro-anaerobic zone and carbon supply inside the coated biofilm, the high DO level in the PBS-CW increased the abundance of the nitrifying bacteria (amoA and nxrA), denitrifying bacteria (narG, nirK, nirS, and nosZ), and even anammox bacteria (anammox 16s rRNA). These features are beneficial to many microbial processes which require the simultaneous aerobic, anoxic, and anaerobic environment.

Intensified nutrients removal in constructed wetlands by integrated Tubifex tubifex and mussels: Performance and mechanisms.

The synergy of Tubifex tubifex (T. tubifex) and mussels on SFCWs (named SFCW-MT) performance was well studied in laboratory throughout a year. The SFCW-MT were steady operated with high TN and TP treatment, with the removal efficiencies of 37.85 ±â€¯5.22% and 39.26 ±â€¯5.20% even in winter. The mussels had excellent NH4-N removal efficiency, and avoid the shortage of NH4-N removal with T. tubifex in winter. Simultaneously, the SFCW-MT improved the NO3-N treatment by 51% than that in control group. The plant growth was improved in SFCW-MT, which reflected in the improvement of total chlorophyll contents and plant heights. The N and P absorbed by wetland plants and adsorbed by substrate were both increased with mussels. Microbial analysis results revealed that, the mussels could keep the abundance of nitrifiers despite the negative effect of T. tubifex. On that basis, the improved proportions of denitrifiers (Firmicutes) have a significantly recognized role in NO3-N transformation in SFCW-MT. The gut and membrane sections of mussels, as well as T. tubifex, also has proportions of denitrifiers and part of nitrifiers, and thus changed the microbial community in substrate. This evidence indicated that the co-existence of T. tubifex and mussels have potential application for simultaneous removal of NH4-N and NO3-N in CWs.

Tetrabromobisphenol A alters soil microbial community via selective antibacterial activity.

Tetrabromobisphenol A (TBBPA) is the most widely used brominated flame retardant. Most studies regarding TBBPA have concentrated on its occurrence, distribution, toxicity and degradation in the environment. However, little is known about its ecological effects on soil microbial communities. In this study, we investigated the effect of TBBPA on soil microbial community. Overall, the data suggested that the growth and composition of soil microorganisms were correlated to the TBBPA concentration and exposure time. Phospholipid-derived fatty acid analysis (PLFAs) showed that significant microbial growth inhibitions were 46.1% and 46.9% in 40 mg/kg TBBPA-treated soils after 45-day incubation under aerobic and anaerobic conditions, respectively. Results of PLFAs and llumina sequencing indicated that TBBPA mainly inhibited Gram-positive bacteria, but not Gram-negative bacteria. The selective antibacterial activity of TBBPA toward Gram-positive bacteria was further confirmed in pure bacteria cultures. These data suggested that, in addition to their effect on microbial growth and composition, TBBPA may affect the microbial ecology. Additional research should be carried out to identify the ecological risk of TBBPA in soil.

Multiregion sequencing reveals the intratumor heterogeneity of driver mutations in TP53-driven non-small cell lung cancer.

Intratumor heterogeneity (ITH) in non-small cell lung cancer (NSCLC) may account for resistance after a period of targeted therapies because drugs destroy only a portion of tumor cells. The recognition of ITH helps identify high-risk patients to make effective treatment decisions. However, ITH studies are confounded by interpatient heterogeneity in NSCLC and a large amount of passenger mutations. To address these issues, we recruited NSCLC patients carrying TP53 mutations and selected driver mutations within recurrently mutated genes in NSCLC. A total of 12-paired normal-tumor tissues were subjected to whole-genome/whole-exome sequencing. From these, 367 non-silent mutations were selected as driver mutations and deeply sequenced in 61 intratumoral microdissections. We identified a universal prevalence of heterogeneity in all 12 tumors, indicating branched evolution. Although TP53 mutations were observed in single biopsy of all 12 tumors, most tumors consist of both TP53 mutated and non-mutated cells in separate regions within the same tumor. This suggests the late molecular timing of the acquisition of TP53 mutations; therefore, the detection of TP53 mutations in a single biopsy may simply not reflect the early malignant potential. In addition, we identified regions of loss of heterozygosity surrounding TP53 and CDKN2A mutations in tumor 711, which also exhibited heterogeneity in different regional samples. Because the ITH of driver mutations likely has clinical consequences, further efforts are needed to limit the impact of ITH and to improve therapeutic efficiency, which will benefit NSCLC patients receiving targeted treatments.

Periplasmic Nicotine Dehydrogenase NdhAB Utilizes Pseudoazurin as Its Physiological Electron Acceptor in Agrobacterium tumefaciens S33.

Agrobacterium tumefaciens S33 can grow with nicotine as the sole source of carbon, nitrogen, and energy via a novel hybrid of the pyridine pathway and the pyrrolidine pathway. Characterization of the enzymes involved in the hybrid pathway is important for understanding its biochemical mechanism. Here, we report that the molybdenum-containing nicotine dehydrogenase (NdhAB), which catalyzes the initial step of nicotine degradation, is located in the periplasm of strain S33, while the 6-hydroxynicotine oxidase and 6-hydroxypseudooxynicoine oxidase are in the cytoplasm. This is consistent with the fact that NdhA has a Tat signal peptide. Interestingly, an open reading frame (ORF) adjacent to the ndhAB gene was verified to encode a copper-containing electron carrier, pseudoazurin (Paz), which has a signal peptide typical of bacterial Paz proteins. Both were transported into the periplasm after being produced in the cytoplasm. We purified NdhAB from the periplasmic fraction of strain S33 and found that with Paz as the physiological electron acceptor, NdhAB catalyzed the hydroxylation of nicotine at a specific rate of 110.52 ± 8.09 µmol · min-1 · mg of protein-1, where the oxygen atom in the hydroxyl group of the product 6-hydroxynicotine was derived from H2O. The apparent Km values for nicotine and Paz were 1.64 ± 0.07 µM and 3.61 ± 0.23 µM, respectively. NAD(P)+, O2, and ferredoxin could not serve as electron acceptors. Disruption of the paz gene disabled the strain for nicotine degradation, indicating that Paz is required for nicotine catabolism in the strain. These findings help our understanding of electron transfer during nicotine degradation in bacteria.IMPORTANCE Nicotine is a toxic and addictive N-heterocyclic aromatic alkaloid produced in tobacco. Its catabolism in organisms and degradation in tobacco wastes have become major concerns for human health and the environment. Bacteria usually decompose nicotine using the classical strategy of hydroxylating the pyridine ring with the help of activated oxygen by nicotine dehydrogenase, which binds one molybdopterin, two [2Fe2S] clusters, and usually one flavin adenine dinucleotide (FAD) as well. However, the physiological electron acceptor for the reaction is still unknown. In this study, we found that the two-component nicotine dehydrogenase from Agrobacterium tumefaciens S33, naturally lacking an FAD-binding domain, is located in the periplasmic space and uses a copper-containing electron carrier, pseudoazurin, as its physiological electron acceptor. We report here the role of pseudoazurin in a reaction catalyzed by a molybdopterin-containing hydroxylase occurring in the periplasmic space. These results provide new biochemical knowledge on microbial degradation of N-heterocyclic aromatic compounds.

Performance and mechanism of triclosan removal in simultaneous nitrification and denitrification (SND) process under low-oxygen condition.

Wastewater treatment under low dissolved oxygen (DO) conditions is promising for its low energy consumption. However, the removal process of some organic micropollutants, such as triclosan (TCS), could be inhibited under anaerobic conditions. So, it is worth investigating the TCS removal performance at low-oxygen condition. In this study, simultaneous nitrification and denitrification (SND) process, with DO ranging from 0.30 to 0.80 mg L-1, was chosen to investigate. Results showed that the water quality of the effluent was deteriorated after TCS addition at the beginning, with removal efficiency of NH4+-N dropped from almost 100 ± 0.70 to 88.30 ± 0.98% and COD decreased from 95.15 ± 1.55 to 65.81 ± 2.42 %. However, the performance recovered from the 3rd day and almost stabilized on the 14th day with the removal efficiencies of NH4+-N were over 98.00 ± 0.60 %, and COD was above 94.00 ± 1.70 % in effluent. Besides, TCS removal efficiencies were more than 93.00 %, and the contributions for TCS removal by the water effluent, sludge sorption, and other effects including biodegradation were 6.46 ± 2.25, 16.27 ± 3.30, and 77.27 ± 4.45 %, respectively. Although the results of absolute abundances of related genes showed no difference (P > 0.05), Illumina MiSeq sequencing analysis presented the variation of microbial community after TCS addition, in which T-45 had the highest Shannon and Simpson diversity index, followed by T-0 and T-2. Relative abundances of alpha and beta-Proteobacteria, which were related to TCS biodegradation, were increased. Compared with Bacteroidetes in T-0, the abundance of Bacteroidetes took up more than 15.6 % in T-45, which should play a more important role under low-oxygen conditions with TCS addition.

PGK1 Drives Hepatocellular Carcinoma Metastasis by Enhancing Metabolic Process.

During the proliferation and metastasis, the tumor cells prefer glycolysis (Warburg effect), but its exact mechanism remains largely unknown. In this study, we demonstrated that phosphoglycerate kinase 1 (PGK1) is an important enzyme in the pathway of metabolic glycolysis. We observed a significant overexpression of PGK1 in hepatocellular carcinoma tissues, and a correlation between PGK1 expression and poor survival of hepatocellular carcinoma patients. Also, the depletion of PGK1 dramatically reduced cancer cell proliferation and metastasis, indicating an oncogenic role of PGK1 in liver cancer progression. Further experiments showed that PGK1 played an important role in MYC-induced metabolic reprogramming, which led to an enhanced Warburg effect. Our results revealed a new effect of PGK1, which can provide a new treatment strategy for hepatocellular carcinoma, as PGK1 is used to indicate the prognosis of hepatocellular carcinoma (HCC).

Genome-wide association study identifies a novel susceptibility gene for serum TSH levels in Chinese populations.

Thyroid-stimulating hormone (TSH) is a sensitive indicator of thyroid function. High and low TSH levels reflect hypothyroidism and hyperthyroidism, respectively. Even within the normal range, small differences in TSH levels, on the order of 0.5-1.0 mU/l, are associated with significant differences in blood pressure, BMI, dyslipidemia, risk of atrial fibrillation and atherosclerosis. Most of the variance in TSH levels is thought to be genetically influenced. We conducted a genome-wide association study of TSH levels in 1346 Chinese Han individuals. In the replication study, we genotyped four candidate SNPs with the top association signals in an independent isolated Chinese She cohort (n = 3235). We identified a novel serum TSH susceptibility locus within XKR4 at 8q12.1 (rs2622590, Pcombined = 2.21 × 10(-10)), and we confirmed two previously reported TSH susceptibility loci near FOXE1 at 9q22.33 and near CAPZB at 1p36.13, respectively. The rs2622590_T allele at XKR4 and the rs925489_C allele near FOXE1 were correlated with low TSH levels and were found to be nominally associated to patients with papillary thyroid carcinoma (PTC) (OR = 1.41, P= 0.014 for rs2622590_T, and OR = 1.61, P= 0.030 for rs925489_C). The rs2622590 and rs925489 genotypes were also correlated with the expression levels of FOXE1 and XKR4, respectively, in PTC tissues (P = 2.41 × 10(-4) and P= 0.02). Our findings suggest that the SNPs in XKR4 and near FOXE1 are involved in the regulation of TSH levels.

Nicotine Dehydrogenase Complexed with 6-Hydroxypseudooxynicotine Oxidase Involved in the Hybrid Nicotine-Degrading Pathway in Agrobacterium tumefaciens S33.

Nicotine, a major toxic alkaloid in tobacco wastes, is degraded by bacteria, mainly via pyridine and pyrrolidine pathways. Previously, we discovered a new hybrid of the pyridine and pyrrolidine pathways in Agrobacterium tumefaciens S33 and characterized its key enzyme 6-hydroxy-3-succinoylpyridine (HSP) hydroxylase. Here, we purified the nicotine dehydrogenase initializing the nicotine degradation from the strain and found that it forms a complex with a novel 6-hydroxypseudooxynicotine oxidase. The purified complex is composed of three different subunits encoded by ndhAB and pno, where ndhA and ndhB overlap by 4 bp and are ∼26 kb away from pno. As predicted from the gene sequences and from chemical analyses, NdhA (82.4 kDa) and NdhB (17.1 kDa) harbor a molybdopterin cofactor and two [2Fe-2S] clusters, respectively, whereas Pno (73.3 kDa) harbors an flavin mononucleotide and a [4Fe-4S] cluster. Mutants with disrupted ndhA or ndhB genes did not grow on nicotine but grew well on 6-hydroxynicotine and HSP, whereas the pno mutant did not grow on nicotine or 6-hydroxynicotine but grew well on HSP, indicating that NdhA and NdhB are responsible for initialization of nicotine oxidation. We successfully expressed pno in Escherichia coli and found that the recombinant Pno presented 2,6-dichlorophenolindophenol reduction activity when it was coupled with 6-hydroxynicotine oxidation. The determination of reaction products catalyzed by the purified enzymes or mutants indicated that NdhAB catalyzed nicotine oxidation to 6-hydroxynicotine, whereas Pno oxidized 6-hydroxypseudooxynicotine to 6-hydroxy-3-succinoylsemialdehyde pyridine. These results provide new insights into this novel hybrid pathway of nicotine degradation in A. tumefaciens S33.

Heterologous overproduction of 2[4Fe4S]- and [2Fe2S]-type clostridial ferredoxins and [2Fe2S]-type agrobacterial ferredoxin.

Ferredoxins are small, acidic proteins containing iron-sulfur clusters that are widespread in living organisms. They play key roles as electron carriers in various metabolic processes, including respiration, photosynthesis, fermentation, nitrogen fixation, carbon dioxide fixation, and hydrogen production. However, only several kinds of ferredoxins are commercially available now, greatly limiting the investigation of ferredoxin-related enzymes and metabolic processes. Here we describe the heterologous overproduction of 2[4Fe4S]- and [2Fe2S]-type clostridial ferredoxins and [2Fe2S]-type agrobacterial ferredoxin. Adding extra iron and sulfur sources to the medium in combination with using Escherichia coli C41(DE3) harboring pCodonplus and pRKISC plasmids as host greatly enhanced iron-sulfur cluster synthesis in the three ferredoxins. After induction for 12 h in terrific broth and purification by affinity chromatography and anion exchange chromatography, approximately 3.4 mg of streptavidin (Strep)-tagged and 3.7 mg of polyhistidine (His)-tagged clostridial 2[4Fe4S] ferredoxins were obtained from 1 l of culture. Excitingly, after induction for 24 h in terrific broth, around 40 mg of His-tagged clostridial [2Fe2S] and 23 mg of His-tagged agrobacterial [2Fe2S] ferredoxins were purified from 1 l of culture. The recombinant ferredoxins apparently exhibited identical properties and physiological function to native ferredoxins. No negative impact of two different affinity tags on ferredoxin activity was found. In conclusion, we successfully developed a convenient method for heterologous overproduction of the three kinds of ferredoxins with satisfactory yields and activities, which would be very helpful for the ferredoxin-related researches.

Triclosan removal in wetlands constructed with different aquatic plants.

Triclosan (TCS) is widely used in consumer products as an antimicrobial agent. Constructed wetlands have the potential for TCS removal, but knowledge about the relative importance of sediment, plants, and microbes is limited. TCS removal performance was investigated in well-operated constructed wetlands planted with three different types of aquatic plants: emergent Cattail (C-T), submerged Hornwort (H-T), and floating Lemnaminor (L-T). Results showed that the TCS removal efficiencies from water were all greater than 97 %. Maximal TCS adsorption to sediment in the C-T wetland (13.8 ± 0.6 ng/g) was significantly lower than in the H-T wetland (21.0 ± 0.3 ng/g) or the L-T wetland (21.4 ± 0.6 ng/g). The maximal TCS concentrations in plants were 5.7 ± 0.2 and 7.2 ± 0.5 µg/g for H-T and L-T, respectively, and it was below the minimal detection limit (MDL) in C-T. Deep 16S rRNA gene sequencing results revealed that C-T wetland had the highest community richness and diversity. Some bacteria, like beta-Proteobacteria, gamma-Proteobacteria, and Bacteroidetes were detected and might have significant correlations with TCS degradation. Overall, with regard to soils, plants, and microorganism, accumulation in sediment and plants in H-T and L-T was high, while in C-T biodegradation likely played an important role.