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The promotional effects of sodium on Fe5C2 catalysts for CO2 hydrogenation were systematically investigated by a synergistic combination of density functional theory (DFT) calculations and experime...

Structure-based prediction of a compound's potential sites of metabolism (SOMs) mediated by cytochromes P450 (CYPs) is highly advantageous in the early stage of drug discovery. However, the SOMs prediction accuracy could be influenced by several factors. CYP2C9 is one of the major drug metabolizing enzymes in humans and is responsible for the metabolism of ~13% of clinically used drugs. In this study, we systematically evaluated the effects of protein crystal structural models, scoring functions, heme forms, conserved active site water molecule, and protein flexibility on SOMs prediction of CYP2C9 substrates. Our results demonstrated that ChemScore and GlideScore outperformed four other scoring functions including Vina, GoldScore, ChemPLP, and ASP on average. The performance of the crystal structural models with the pentacoordinated heme was generally superior to that of the hexacoordinated iron-oxo heme (referred to as Compound I) models. The inclusion of the conserved active site water molecule improved the prediction accuracy by GlideScore but reduced the accuracy by ChemScore. In addition, the effect of the conserved water on SOMs prediction was found to be dependent on the receptor model and the substrate. We further found that one of snapshots from molecular dynamics simulations on the apo form can improve the prediction accuracy when compared to the crystal structural model.

Photothermal therapy (PTT) based on semiconducting polymer nanoparticles (SPNs) is a promising strategy to treat solid tumors, but its ability to combine with chemotherapy for immune remodeling to efficiently suppress metastatic cancers has rarely been studied. Here, we demonstrate that PTT combined with chemotherapy can efficiently elicit immunity to suppress metastatic tumor growth. Specifically, we rationally designed a new SPN (PDPSe NPs) as a photothermal agent for PTT with a large mass extinction coefficient in the near-infrared region (e.g., 44.9 L g-1 cm-1 at 808 nm), high photothermal conversion efficiency (62.5%) and excellent biocompatibility. A hypoxia-activated anti-tumor drug, tirapazamine (TPZ), was selected for chemotherapy. Strikingly, the combination therapy not only induced tumor cell death in the primary tumor, but also effectively suppressed the growth of distant tumors (mimicking metastatic tumors) without PTT. Importantly, the combined therapies exhibit synergistic effects on immune remodeling. Immunofluorescence data suggest that the inhibition of metastatic tumor growth is attributed to the immune remodeling triggered by PTT and chemotherapy. This work demonstrates a new paradigm of utilizing PTT together with hypoxia-activated drugs to effectively retard metastatic tumor growth.

Background and Purpose: The results regarding the independent association between homocysteine (Hcy) levels and post-stroke cognitive impairment (PSCI) were inconsistent. The effect of age on this association has yet to be explored. This study aims to determine the relationship between Hcy levels, age, and cognitive impairment in a post-stroke population.Methods: A total of 592 patients with acute ischemic stroke (AIS) completed follow-up. Serum Hcy levels were measured enzymatically by spectrophotometry within 24 h of admission. Cognitive function was evaluated by the Mini-Mental State Examination (MMSE) 1 month after stroke, and the scores ≤ 24 were considered as cognitive impairment. Our study was dichotomized into two groups by a cut-off of 65 years. Multivariate logistic regression models were used to determine the association between baseline Hcy levels and cognitive impairment.Results: According to the MMSE score, 317 (53.5%) patients had cognitive impairment. Patients with higher levels of Hcy were more prone to have cognitive impairment 1 month after stroke than patients with lower levels of Hcy (p < 0.001). The optimal cut-off points of Hcy level (μmol/L) were (T1) ≤ 8, (T2) 8–12, and (T3) ≥ 12. After adjusting for confounding factors, the multivariate regression analysis showed that the third Hcy tertile was independently associated with cognitive impairment [odds ratio (OR) = 2.057, 95% confidence interval (CI) = 1.133–3.735, p = 0.018). A stronger association [T2 (OR = 2.266, 95% CI = 1.042–4.926, p = 0.039); T3 (OR =3.583, 95% CI = 1.456–8.818, p = 0.005)] was found in the younger group. However, the independent association was not confirmed in the older group.Conclusions: Elevated Hcy levels in the acute phase of ischemic stroke were independently associated with cognitive impairment in a post-stroke population. Furthermore, the association was age-dependent and more meaningful in a younger population aged below 65. So, Hcy levels in patients with stroke should be well-monitored, especially in younger patients.

Random migration often plays important roles in soil microbial community assembly. However, it remains unexplored that whether its role is affected by anthropogenic disturbances, and whether the effects of different disturbances are consistent. Here, we report a novel and general pattern that different disturbances consistently restrain the role of migration in soil bacterial community assembly. We mimicked 16 types of anthropogenic disturbances relevant to a Eurasian steppe ecosystem, including removing different number of plant functional groups, mowing, adding nitrogen, adding phosphorus, watering, warming, and some of their combinations. The estimated migration rates in soil bacterial community assembly with Stephen Hubbell’s neutral model decreased by 0.19–6.29% under these disturbances. The relative change in migration rate was correlated with the deterministic change in community composition and the relative change in soil pH. Our results suggested that the intensifying anthropogenic disturbances may restrain the random dispersal, colonization and distribution of soil microorganisms.

Present study reports a controllable phase transformation of nickel (Ni) from amorphous to cubic crystal structures on tungsten (W) substrate by electrodeposition. X-ray powder diffraction, scanning electron microscopy, transmission electron microscopy and energy dispersive spectroscopy were used to characterize the microstructure, micro-constituents and surface morphology of as-prepared Ni. The microstructure of Ni was strongly affected by the applied overpotential and deposition time. It is demonstrated that by controlling these two parameters either amorphous or cubic crystal structure of Ni on the W substrate could be obtained. The crystallization mechanism is discussed based on Gibbs crystal growth theory and Ostwald’s rule. It is concluded that W substrate, acting as a heat sink, can effectively promote the thermal stability of amorphous Ni, based on the data from differential scanning calorimetry and Kissinger’s model. This work contributes to the elucidation of the crystallization mechanism of Ni on W powder substrates, and proves that, better than alloying with other elements, incorporating powder substrates will significantly improve the crystallization temperature, hence the thermostability of amorphous Ni.

Electroless nickel/nickel-boron (Ni/Ni-B), Ni/alkaline Ni-P and Ni/acidic Ni-P double-layer coatings were deposited on AZ31 magnesium (Mg) alloy prepared by environmental fluoride-free solution under low temperatures. The morphologies, compositions, and crystal structures of three different double-layer coatings were characterized by field emission scanning electron microscopy (FESEM), energy dispersive X-ray spectroscopy (EDS) and X-ray diffraction (XRD), respectively. The corrosion resistance of the different double-layer coatings on Mg alloy surface was studied by electrochemical polarization and electrochemical impedance spectroscopic (EIS) tests, which indicated that the Ni/Ni-B coating presented the more superior corrosion resistance than the Ni/alkaline Ni-P and Ni/acidic Ni-P coatings. Meanwhile, the Ni/Ni-B coating exhibited good repeatability through the electrochemical polarization tests for three Ni/Ni-B samples that prepared under the same processing conditions. In addition, the optimal plating time for the Ni-B coating was further determined by electrochemical polarization tests of samples with different plating times.

Purpose Nitrogen deposition at rate not more than 50 kg ha−1 year−1 is generally considered to stimulate soil nitrifying and denitrifying microorganisms via increases in soil nitrogen content. However, this phenomenon in alpine ecosystems remains largely untested. Methods We conducted an 8-year nitrogen deposition experiment on the Qinghai–Tibetan Plateau, with four nitrogen deposition rates of 10 (atmospheric deposition), 20, 30, and 50 kg ha−1 year−1. Results The abundances of two nitrifying genes and four denitrifying genes and the N2O emission rate initially increased and subsequently decreased as the nitrogen deposition rate increased. The observed decrease in these indices at the rate of 50 kg ha−1 year−1 was caused by the toxicity of excessive NH4+. Conclusions Our study demonstrates the vulnerability of alpine microorganisms under global changes.

Gender-specific differences were found in serum uric acid (SUA) levels and the risk of isolated distal deep vein thrombosis (IDDVT). This study aimed to explore the association among gender, SUA, and IDDVT in stroke patients.Finally, 3404 patients were recruited and divided into two groups: IDDVT (n = 1233) and Non-IDDVT (n = 2171) groups. Propensity score matching (PSM) was conducted to match the patients. Binary logistic regression was adopted to explore the association between SUA and IDDVT, with the SUA divided into quartiles. After PSM, 975 patients were included in each group. Non-IDDVT group had a larger proportion of male than IDDVT group (64.9% vs. 52.7%, p 0.001). Moreover, males showed higher SUA levels than females (316.7 ± 102.1 vs. 261.8 ± 94.0 μmol/L, t = 12.1, p 0.001). The highest quartile of SUA (≥346 μmol/L) showed a lower risk of IDDVT (OR = 0.629, p = 0.001), while the lowest quartile (≤225 μmol/L) showed a higher risk of IDDVT (OR = 1.361, p = 0.022).In patients with stroke, SUA played a protective role in IDDVT. Females had a higher risk of IDDVT, which may be owing to the lower SUA levels than males. In clinical practice, more attention should be paid to the risk of IDDVT in females, especially those with lower SUA levels.

High-quality monodispersed copper nanowires with an ultrathin diameter of 13.5 nm, lengths up to 30 μm (aspect ratio >104) were successfully synthesized by a facile and controllable hydrothermal reduction procedure. The synthesis utilized glucose in the presence of hexadecylamine (HDA) and octadecylamine (ODA) as the capping agents. The copper decahedra nanoparticles with a low-surface-energy {111} plane formed pentatwinned one-dimensional nanowires, which was exactly verified by selected-area electron diffraction. Furthermore, the diameter and relative film conductivity of copper nanowires are sensitive to the HDA/ODA molar ratio. The conductor film made of the high-quality and ultrathin copper nanowires shows high transmittance and low resistance (83.83%, 61 Ω/□), exhibiting great potential in the applications of nanofabrication, transparent and flexible conductors, organic light-emitting diodes, and more.

The performance of Fe-based catalysts has found to be remarkably improved by the addition of ZnO in the reaction of CO2 hydrogenation to linear α-olefins (LAO). The space time yield (STY) of C4-20 α-olefins for bimetallic Fe2Zn1 is 2.4 times of the Fe alone; while a C4-20 olefins selectivity of 60.7 % with an 89.3 % LAOs/olefins ratio in C4-20 alkenes at a CO2 conversion of 43.5 % was achieved as well. Using in situ X-ray photon spectroscopy (XPS), we have proved that the chemical state of Zn varied between 0 and +2 (Znδ+), accompanying by electronic donating from ZnOx to Fe. Focusing on electronic effects, we revealed the structure evolution during the induction period. The introduction of ZnO changed the ratios of FeCx/FeOx during the reaction. This study uncovered the chemical valence of Zn for CO2 to LAOs and its effects on the structure of active sites and the performance of Fe5C2-ZnO catalysts.

Agonistic behaviour is common in an encounter between two crustaceans. It often causes limb disability and consumes a lot of energy, which is harmful for the growth and survival of commercially important crustaceans. In the present study, we mainly focused on the agonistic behaviour of the Chinese mitten crab, Eriocheir sinensis, which is an important species of the aquaculture industry in China. We recorded agnostic behaviour with a high-definition camera and preliminarily evaluated the role of serotonin (5-HT) or dopamine (DA)-cyclic adenosine monophosphate (cAMP)-protein kinase A (PKA) pathway and eyestalk in the behaviour. The results showed that agonistic behaviour in E. sinensis consisted of three stages: approach, contact and fight. We found that the number of fights and cumulative time of fight were significantly higher in the male vs. male group than in the female vs. female and female vs. male groups (P

Ammonia-oxidizers play an essential role in nitrogen (N) transformation and nitrous oxide (N2O) emission in forest soils. It remains unclear if ammonia-oxidizers affect interaction between methane (CH4) uptake and N2O emission. Our specific goal was to test the impacts of changes in ammonia-oxidizing communities elicited by N enrichment on soil CH4 uptake and N2O emission. Based on a field experiment, two-forms (NH4Cl and NaNO3) and two levels (40 and 120 kg N ha−1 yr−1) of N were applied in the subtropical plantation forest of southern China. Soil CH4 and N2O fluxes, the abundance and structure of ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB) communities were measured using static chamber-gas chromatography, quantitative PCR (qPCR), and terminal-restriction fragment length polymorphism (T-RFLP). Nitrogen addition tended to inhibit soil CH4 uptake, but significantly promoted soil N2O emission; moreover, these impacts were more significant with NH 4 + − N than with NO 3 − − N addition. NH4Cl addition significantly changed ammonia-oxidizer abundance with an increase in AOA and a decrease in AOB. Nitrogen additions significantly decreased the relative abundance of 329 bp and 421 bp of archaeal amoA gene. Negative relationships occurred between soil CH4 uptake and AOA abundance and between soil CH4 uptake and AOA/AOB ratio; however, a positive relationship was found between soil N2O emission and AOA abundance. These results indicate that a shift in abundance and composition of ammonia-oxidizing communities is closely linked to changes in soil CH4 uptake and N2O emission under N enrichment. Furthermore, AOA communities play a contrasting role from AOB communities for regulating the fluctuation between soil CH4 and N2O fluxes.

Background and aims Deposited NH 4 + and NO 3 − differently affect soil carbon (C) and nitrogen (N) cycles due to their contrasting actions in terrestrial ecosystems. However, little information on the effects of exogenous NH 4 + and NO 3 − inputs on the exchange of greenhouse gases (GHGs) from the subtropical plantation soils as well as their contribution to global warming is available to date. Methods Based on a field experiment, two-form (NH 4 Cl and NaNO 3 ) and two-level (40 and 120 kg N ha −1 yr −1 ) of N addition, in a slash pine plantation of southern China, we investigated soil CO 2 , CH 4 and N 2 O fluxes and related auxiliary variables (soil temperature and moisture) twice a week using static chamber–gas chromatography. The total global warming potential (GWP) of soil GHG fluxes and N 2 O emission factor (EF) were calculated. Results Low level of NaNO 3 addition significantly increased cumulative annual soil CO 2 emission by 33.7%. N addition significantly promoted annual soil N 2 O emission by 2.4–6.9 folds; moreover, ammonium-N addition had a greater promotion to soil N 2 O emission than nitrate-N addition. However, short-term N addition did not change soil CH 4 uptake. Also, soil CO 2 and N 2 O fluxes were positively correlated with soil temperature and moisture, while soil CH 4 uptake was only driven by soil moisture. Overall, elevated N addition increased the total GWP, and changed the temperature sensitivity (Q 10 ) of soil CO 2 and N 2 O fluxes. Conclusions These results suggest that chronic atmospheric N deposition changes soil-atmospheric GHG fluxes in the subtropical plantation of southern China depending on the levels and forms of N input, and would exacerbate global warming.

Predominant loss of neutral CO 2 has been observed under conditions of low-energy collision-induced dissociation from a prototypical molecular radical cation of the tripeptide aspartylglycylarginine ([DGR] + ). The decarboxylation occurs mainly from the side chain of the aspartic acid residue and partially from the C-terminal carboxyl group. The structural and mechanistic features that facilitate CO 2 loss from the Asp side chain of [DGR] + and its chemically modified analogs incorporating methylation have been elucidated using a combination of Rice–Ramsperger–Kassel–Marcus modeling and density functional theory at the B3LYP/6-31++G(d,p) level. Current mechanistic investigations suggest that the loss of CO 2 from the side chain of the aspartic acid residue involves hydrogen atom transfer from its carboxyl oxygen atom in conjunction with α-centered radical transfer to the β-centered radical on the aspartic acid side chain. Minor CO 2 loss from the C-terminal carboxyl group occurs through the [DG α R] + isomer, with the radical migrating to the α-carbon of the middle Gly residue. Barriers against the CO 2 loss from the side chain of the aspartic acid residue and from the C-terminus of [DG α R] + are approximately 30 and 36 kcal mol −1 , respectively.

Knowledge about how Eriocheir sinensis interacts with microorganisms in its ambient environment is still lacking. Using RNA-Seq, we determined the most conserved genes and pathways compared with other animals and detected highly-induced immune genes in E. sinensis hemocytes post-in vivo challenge with different microbial derivatives. In total, 33.2 million high-quality reads were generated and assembled into 177,679 contigs. Completeness assessment and functional annotation were performed. Lipopolysaccharide, peptidoglycan, and β-1, 3-glucan stimulation induced 373, 173, and 108 differentially expressed (DE) transcripts, respectively. GO terms such as 'G-protein-coupled receptor binding', 'negative regulation of mitogen-activated protein kinase activity', and 'positive regulation of blood circulation' were enriched in the DE transcripts. Quantitative real-time PCR validated the data for selected genes. Our data contribute to understanding the immune defense mechanism in E. sinensis and the development of the innate immune system, thereby providing insights into disease control and prevention in aquaculture.

This study investigated the influence of L-tryptophan (L-trp) on the survival, immune response and gut microbiota of the Chinese mitten crab, Eriocheir sinensis (with an average weight of 16.58 ± 2.20 g). After 30 days of feeding with diets supplemented with L-trp at 0.36%, 0.47%, 0.73% and 1.05% (groups 1, 2, 3 and 4, respectively), the survival rate and bacterial challenge (Aeromonas hydrophila) were evaluated, the activities of antioxidant and phosphatase enzymes in the serum were assessed, and the gut microbiota were measured via high-throughput Illumina sequencing. The results showed that the supplementation of L-trp significantly improved the survival rate of crabs (P 0.05). After feeding for 7 days, it was observed that a high L-trp diet significantly increase the survival rate relative to a basal diet after a 96-h post-challenge with A. hydrophila (P 0.05). The activity of CAT and AKP in the serum were increased by the addition of L-trp. The activity of CAT and AKP in the serum in group 4 were higher than those in group 1 (P 0.05). Furthermore, we observed that adjunction of the L-trp can significantly increase the richness and diversity of the gut microbiota. The dominant phylum in the intestine of the Chinese mitten crab were Tenericutes, Proteobacteria, Firmicutes, Chloroflexi and Actinobacteria. The L-trp in the diets increased the richness of Proteobacteria, Firmicutes and Actinobacteria in the intestine significantly. These bacteria were all dominant bacteria and had a specific role in promoting the immunity of E. sinensis. Therefore, it could be inferred that L-trp supplementation is beneficial in the diet of E. sinensis. Based in these results, the dietary 0.47% or 0.73%L-trp supplemented is found to be optimum to improve E. sinensis survival.

GNE myopathy is a rare autosomal recessive distal myopathy caused by mutations in UDP-N-acetylglucosamine 2-epimerase/N-acetylmannosamine kinase (GNE), the bi-functional enzyme critical for sialic acid biosynthesis. In this study, we summarized the clinical features, pathological characteristics, and genetic profiles of 46 GNE patients. The clinical and mutational profile of 54 previously reported Chinese patients were also reviewed. A total of 21 novel mutations, including a gross deletion spanning exon 1-2 and a retrotransposon insertion were found in our cohort, enlarging the spectrum of GNE mutations. The most frequent mutation in Chinese population was D207V, which accounts for 25.5% of total alleles (51/200). The age of onset was much later in the patients carrying D207V compared to other patients, indicated the less deleterious effect of D207V on enzyme activity. GNE myopathy may be overlooked in China with a relatively milder phenotype due to the common mutation D207V.

Melatonin has been identified in a variety of invertebrate species, but its function is not as well understood as in crustaceans. The effects of melatonin on hemolymph glucose levels and tissue carbohydrate metabolism in the Chinese mitten crab, Eriocheir sinensis, were fully investigated in this study. Moreover, whether the eyestalk (an important endocrine center in invertebrate species) involves in this process or not, also were clarified. Analysis revealed that eyestalk ablation, especially bilateral, caused a significant decrease in the hemolymph glucose level. Moreover, injection of melatonin induced hyperglycemia in a dose-dependent manner both in intact and ablated crabs. Based on the expression of CHH mRNA in the 10 different tissues, eyestalk, thoracic ganglion, intestinal tract and hemolymph were selected to estimate the effect of melatonin on the expression of CHH mRNA. Bilateral eyestalk ablation caused a significant increase in the expression of CHH mRNA in the thoracic ganglion, intestinal tract and hemolymph compared with the controls. In addition, injection of melatonin into intact or ablated crabs elevated the CHH mRNA level in the eyestalk, thoracic ganglion and intestinal tract tissues compared with controls. The hemolymph CHH mRNA after melatonin injection was elevated only in ablated crabs. Administration of melatonin resulted in a significant decrease in total carbohydrates and glycogen levels with an increase in phosphorylase activity levels in the hepatopancreas and muscle in intact and ablated crabs. Our findings demonstrated that melatonin can induce hyperglycemic effects in both intact and ablated crabs, suggesting that this effect is probably not mediated solely via eyestalk.

Dopamine (DA) plays a modulatory role in numerous physiological processes such as light adaptation and food intake, and exerts these functions through DA receptors (DARs). This study presents, for the first time, isolation and characterization of the dopamine receptor 2(DA2 receptor) cDNA from the intestinal tissue of Eriocheir sinensis, an economically important freshwater aquaculture species in China. The DA2 receptor cDNA sequence, which was obtained by rapid amplification of cDNA ends, is 2369bp long, encode peptide of 589 amino acid, and is highly homologous to related sequences in crustaceans. Analysis of the deduced amino acid sequence and the structure of the DA2 indicated that this receptor is a member of the family of G protein-coupled receptors (GPCRs), as it contains seven transmembrane domains and other common signatures of GPCRs. RT-PCR showed that the expression of the DA2 receptor gene was distributed in various tissues, and high expression levels were observed in the cranial ganglia and the thoracic ganglia. Further study of the effect of photoperiod on DA2 expression showed that constant darkness induced a significant increase in DA2 expression in the cranial ganglia. Finally, analysis of DA2 receptor expression under different feeding statuses showed that there was significantly greater expression in the hepatopancreas and intestines after feeding than before feeding, but there were no differences in expression between the before feeding and during feeding periods in either tissue. Our results indicate that the DA2 receptor structurally belongs to the family of G protein-coupled receptors, and that the cranial ganglia are the main tissues in which the DA2 receptor participates in light adaptation during dark hours. In addition, the DA2 receptor in E. sinensis may be involved in the physiological regulation of the hepatopancreas and digestive tract after the ingestion of food. This study provides a foundation for further exploration of the light adaptation and digestive functions of the DA2 receptor in decapods.

Fifteen semi-synthetic derivatives of asiatic acid (AA) have been synthesized and evaluated for their biological activities. The successful modification of these compounds at the C-2, C-3, C-23 and C-28 positions was confirmed using NMR, MS and IR spectra. Further, their anti-tumor effects were evaluated in vitro using different cancer cell lines (HeLa, HepG2, B16F10, SGC7901, A549, MCF7 and PC3), while their anti-angiogenic activities were evaluated in vivo using a larval zebrafish model. Among the derivatives, compounds 4–10 showed more potent cytotoxic and anti-angiogenic effects than AA, while compounds 11–17 had significantly less effects. The new derivative 10 was also included in finished formulations to evaluate its stability using HPLC due to its potential topical use. The derivative 10 had markedly better anti-tumor activities than both AA and other derivatives, with similar stability as its parent compound AA.

Nitrogen (N) deposition has been shown to affect soil carbon (C) and N cycling in subtropical forests; however, the underlying microbial mechanisms are poorly understood. We used patterns of community-level physiological profiles and enzyme activities to assess the relative effects of the addition of four N levels (0, 50, 100, and 150 kg·ha−1·year−1) on the soil microbial community in three forest stands (pine, mixed, and broadleaf forests) in southern China, where the forests have been experimentally manipulated for over 8 years. In pine forests, N50 addition significantly increased microbial biomass carbon (MBC) concentration but decreased soil pH levels. N100 addition significantly increased soil peroxidase activity but decreased soil β-1,4-glucosidase activity. In broadleaf forests, N addition increased soil dissolved organic carbon (DOC) concentration and polyphenol oxidase activity but decreased soil MBC concentration and soil pH levels. N addition also significantly increased soil microbial metabolism activity (expressed as average well color development) in pine forest and broadleaf forest soils. However, the mixed forests responded slowly to N additions and exhibited no significant response of C-utilization profiles and soil enzyme activities. Principal component analysis of C-utilization data separated microbial communities with respect to N addition and forest successional stage. In addition, microbial C utilization was driven by soil pH levels. Although enzyme activities were correlated with soil MBC and microbial biomass nitrogen concentrations, stepwise regression results indicated that soil total carbon contents that were integrated with soil pH levels were key integrators of soil enzyme activities. Our results suggest that soil acidification due to N addition increased soil bacterial C utilization and enzyme activities.

Carbon dioxide (CO2) enrichment and increased nitrogen (N) deposition can change microbial activity and dissolved organic carbon (DOC) turnover, consequently affecting carbon sequestration in soils. However, we do not have much available information on the relationship between soil DOC and microbial activity under CO2 enrichment and N addition in semi-arid agroecosystems. Using free air CO2 enrichment (FACE), soybean and winter wheat were grown in the field under ambient CO2 (350 μmol mol−1) and elevated CO2 (550 μmol mol−1) conditions subjected to two N fertilizer regimes (132 and 306 kg N ha−1 year−1). Rhizosphere soils and bulk soils at three depths, 0–10, 10–20 and 20–40 cm, were collected to determine water extractable organic matter (WEOM) characteristics with fluorescence spectroscopy and parallel factor analyses of excitation/emission matrix, as well as five extracellular enzymes activities. All significant effects were observed in the topsoil (0–10 cm): elevated CO2 decreased water extractable organic carbon concentration of the rhizosphere soils and bulk soils by 8.5 and 10.1 %, respectively. Furthermore, elevated CO2 changed the composition and structure of soil WEOM by increasing the plant- and microbial-derived components in the rhizosphere and solubilizing soil organic matter (SOM). The activities of β-1,4-glucosidase, cellobiohydrolase, phenol oxidase, and peroxidase were stimulated by elevated CO2 in the rhizosphere soils and bulk soils. Our findings suggest that the stimulation of microbial activity elicited by elevated CO2 increased the turnover of labile WEOM and the solubilization of SOM in the topsoils, which could be adverse to the accumulation and stability of soil carbon in the semi-arid agroecosystems in northern China.

Dissolved organic matter (DOM) plays a central role in driving biogeochemical processes in soils, but little information is available on the relation of soil DOM dynamics to microbial activity. The effects of NO3- and NH4+ deposition in grasslands on the amount and composition of soil DOM also remain largely unclear. In this study, a multi-form, low-dose N addition experiment was conducted in an alpine meadow on the Qinghai-Tibetan Plateau in 2007. Three N fertilizers, NH4Cl, (NH4)(2)SO4 and KNO3, were applied at four rates: 0, 10, 20 and 40 kg N ha(-1) yr(-1). Soil samples from surface (0-10 cm) and subsurface layers (10-20 cm) were collected in 2011. Excitation/emission matrix fluorescence spectroscopy (EEM) was used to assess the composition and stability of soil DOM. Community-level physiological profile (CLAP, basing on the BIOLOG Ecoplate technique) was measured to evaluate the relationship between soil DOC dynamics and microbial utilization of C resources. Nitrogen (N) dose rather than N form significantly increased soil DOC contents in surface layer by 23.5%-35.1%, whereas it significantly decreased soil DOC contents in subsurface layer by 10.4%-23.8%. Continuous five-year N addition significantly increased the labile components and decreased recalcitrant components of soil DOM in surface layer, while an opposite pattern was observed in subsurface layer; however, the humification indices (HIX) of soil DOM was unaltered by various N treatments. Furthermore, N addition changed the amount and biodegradability of soil DOM through stimulating microbial metabolic activity and preferentially utilizing organic acids. These results suggest that microbial metabolic processes dominate the dynamics of soil DOC, and increasing atmospheric N deposition could be adverse to the accumulation of soil organic carbon pool in the alpine meadow on the Qinghai-Tibetan Plateau. (C) 2014 Elsevier B.V. All rights reserved.

It is crucial to understand the effects of enhanced nitrogen (N) deposition on soil methane (CH4) uptake to develop a better comprehension of carbon (C) dynamics in terrestrial ecosystems. A two-year field study was conducted to assess the effects of various forms of N (NH4+ and N-3(-)) and associated N deposition rates (0, 10,20 and 40 kg N ha(-1) yr(-1)) on alpine meadow soil CH4 fluxes on the Qinghai-Tibetan Plateau, China. Soil CH4 fluxes, soil temperature, and soil moisture were monitored weekly using the static chamber technique and gas chromatography. Soil inorganic N pools, soil pH and aboveground biomass were measured monthly to examine the key controlling factors of soil CH4 flux. Our results showed that N addition significantly promoted plant growth and changed soil water-filled pore space (WFPS), but did not alter soil inorganic N storages over the short term. Low rates of N addition significantly decreased the seasonal amount of CH4 uptake by 8.6% compared with the control. Soil CH4 fluxes were mainly determined by soil WFPS, followed by inorganic N availability. N addition increased the contribution of soil WFPS, pH and soil NO3- storage. The observed reduction in CH4 uptake caused by N addition may be largely due to a decrease in physical diffusion, as the biochemical inhibition effects on methanotrophic bacteria are minor. These results suggest that soil inorganic N is a regulatory factor of soil CH4 uptake, and its promotion or inhibition to soil CH4 uptake depends on the N status in terrestrial ecosystems. (C) 2013 Elsevier B.V. All rights reserved.

Polycyclic aromatic hydrocarbons (PAHs) are the most common contaminants in the environment. The primary focus on the toxicity of PAHs is their ability to activate the aryl hydrocarbon receptor (AhR)-mediated pathway and lead to carcinogenesis in different organisms. However, the influence of PAHs on the antioxidant system in mammalian systems has received only limited attention. In the present study, we observed that the intraperitoneal injection of 100 mg/kg 3-methylcholanthrene (3MC) into mice significantly increased reactive oxygen species (ROS) levels and malondialdehyde (MDA) contents and decreased glutathione (GSH) contents and the activity of total antioxidant capacity (T-AOC), indicating that serious oxidative stress had been induced in the liver of mice. Then, the oxidative stress signal activated the nuclear factor erythroid 2-related factor 2/antioxidant response element (Nrf2/ARE) pathway by enhancing the mRNA levels of Nrf2, p38, and Erk2. Moreover, the mRNA levels of Nrf2/ARE target genes, including glutathione peroxidase (Gpx), glutathione reductase (GR), glutathione synthetase (GS), NAD(P)H: quinone oxidoreductase 1 (Nqo1), superoxide dismutase 1 (Sod1), and Sod2, increased significantly after treatment with 3MC for 24 hours. The hepatic levels of NQO1 and the activities of GR and GS were also significantly enhanced at 24 hours after 3MC treatment. Because the expression of NQO1 is co-regulated by Nrf2/ARE and AhR/XRE in mammalian tissues, NQO1 may play an important role in protecting against the oxidative stress induced by 3MC. Taken together, our findings suggested that acute exposure to 3MC altered the cellular redox balance in hepatocytes to trigger Nrf2-regulated antioxidant responses, which may represent an adaptive cell defense mechanism against the oxidative stress induced by PAHs. © 2013 Wiley Periodicals, Inc. Environ Toxicol 29: 1399–1408, 2014.

The effects of increasing nitrogen (N) deposition on soil N dynamics and soil acidification in boreal forests are not well understood. Previous studies have shown that changes in N availability could, in turn, affect the magnitude and direction of soil net N mineralization and nitrification rates as well as soil acidification. To better understand the responses of soil inorganic N content and acidification to increased atmospheric N deposition, a cold-temperate coniferous forest in the Daxing’anling region was selected for a low level N addition experiment with different forms of N. It was hypothesized that the soil inorganic N dynamics influenced by plant uptake and soil transformation, immobilization and loss in these N limited boreal forest soils would be sensitive to N addition, even under low level input. It was also hypothesized that the combined effects of NH 4 + – N and NO 3 - – N addition on soil acidification would be different from their individual effects. During the growing season (May to September) of 2010, soil samples of the organic (Oae horizons combined) and mineral (0–30 cm) layers of each plot were collected to measure inorganic N ( NH 4 + – N and NO 3 - – N ) contents and soil pH. The results showed that most of the NH 4 + – N and NO 3 - – N in the cold-temperate coniferous forest soil was accumulated in the organic layer, and decreased with depth. Contrary to expectations, N addition had no effect on the NH 4 + – N content of the organic or mineral soil, while low N addition significantly increased the soil NO 3 - – N content in the mineral horizons. Moreover, the addition of nitrate fertilizers such as KNO3 and NH4NO3 had a strong effect on soil NO 3 - – N content in the top 10 cm of the mineral layer. Both soil NO 3 - – N and NH 4 + – N contents showed obvious seasonal variations, which were consistent with the seasonal pattern of the soil pH values. This was that low and high N addition tended to lead to accumulated soil NO 3 - – N in the peak of the growing season. High N and NH4NO3 addition significantly reduced soil pH in the mineral horizons, and the effects of NH 4 + – N and NO 3 - – N addition on soil acidification were additive, which agreed with the second hypothesis. These results suggest the responses of the cold-temperate coniferous forest to chronic N deposition are more sensitive than other N rich forests. Soil NO 3 - – N and NH 4 + – N dynamics inconsistently respond to N addition in different soil horizons, months, added N levels and N types. The difference in early response to N input between NH 4 + – N and NO 3 - – N could be related to soil N status and differences in the content and properties of N ions. Environmental variables including soil temperature and moisture have the greatest impact on the N dynamics. Soil acidification caused by N addition affects the overall performance of soil inorganic N dynamics including plant uptake and soil N transformation and loss. It should be noted that soil pH in N limited boreal forest soils may be more vulnerable to N deposition, and its critical load may be 30–50 kg N ha−1 year−1. Long-term studies are needed to reveal the responses of N dynamics and soil pH to increased N deposition and to clarify the mechanisms responsible for soil acidification.

To assess the effects of atmospheric N deposition on the C budget of an alpine meadow ecosystem on the Qinghai–Tibetan Plateau, it is necessary to explore the responses of soil-atmosphere carbon dioxide (CO2) exchange to N addition. Based on a multi-form, low-level N addition experiment, soil CO2 effluxes were monitored weekly using the static chamber and gas chromatograph technique. Soil variables and aboveground biomass were measured monthly to examine the key driving factors of soil CO2 efflux. The results showed that low-level N input tended to decrease soil moisture, whereas medium-level N input maintained soil moisture. Three-year N additions slightly increased soil inorganic N pools, especially the soil NH 4 + -N pool. N applications significantly increased aboveground biomass and soil CO2 efflux; moreover, this effect was more significant from NH 4 + -N than from NO 3 − -N fertilizer. In addition, the soil CO2 efflux was mainly driven by soil temperature, followed by aboveground biomass and NH 4 + -N pool. These results suggest that chronic atmospheric N deposition will stimulate soil CO2 efflux in the alpine meadow on the Qinghai–Tibetan Plateau by increasing available N content and promoting plant growth.

Fascinating N-terminal Cα-C bond cleavages in a series of nonbasic tyrosine-containing peptide radical cations have been observed under low-energy collision-induced dissociation (CID), leading to the generation of rarely observed x-type radical fragments, with significant abundances. CID experiments of the radical cations of the alanyltyrosylglycine tripeptide and its analogues suggested that the N-terminal Cα-C bond cleavage, yielding its [x2 + H](•+) radical cation, does not involve an N-terminal α-carbon-centered radical. Theoretical examination of a prototypical radical cation of the alanyltyrosine dipeptide, using density functional theory calculations, suggested that direct N-terminal Cα-C bond cleavage could produce an ion-molecule complex formed between the incipient a1(+) and x1(•) fragments. Subsequent proton transfer from the iminium nitrogen atom in a1(+) to the acyl carbon atom in x1(•) results in the observable [x1 + H](•+). The barriers against this novel Cα-C bond cleavage and the competitive N-Cα bond cleavage, forming the complementary [c1 + 2H](+)/[z1 - H](•+) ion pair, are similar (ca. 16 kcal mol(-1)). Rice-Ramsperger-Kassel-Marcus modeling revealed that [x1 + H](•+) and [c1 + 2H](+) species are formed with comparable rates, in agreement with energy-resolved CID experiments for [AY](•+).

The mobility of the radical center in three isomeric triglycine radical cations[G(*)GG](+), [GG(*)G](+), and [GGG(*)](+) has been investigated theoretically via density functional theory (DFT) and experimentally via tandem mass spectrometry. These radical cations were generated by collision-induced dissociations (CIDs) of Cu(II)-containing ternary complexes that contain the tripeptides YGG, GYG, and GGY, respectively (G and Y are the glycine and tyrosine residues, respectively). Dissociative electron transfer within the complexes led to observation of [Y(*)GG](+), [GY(*)G](+), and [GGY(*)](+); CID resulted in cleavage of the tyrosine side chain as p-quinomethide, yielding [G(*)GG](+), [GG(*)G](+), and [GGG(*)](+), respectively. Interconversions between these isomeric triglycine radical cations have relatively high barriers (or = 44.7 kcal/mol), in support of the thesis that isomerically pure [G(*)GG](+), [GG(*)G](+), and [GGG(*)](+) can be experimentally produced. This is to be contrasted with barriers17 kcal/mol that were encountered in the tautomerism of protonated triglycine [Rodriquez C. F. et al. J. Am. Chem. Soc. 2001, 123, 3006-3012]. The CID spectra of [G(*)GG](+), [GG(*)G](+), and [GGG(*)](+) were substantially different, providing experimental proof that initially these ions have distinct structures. DFT calculations showed that direct dissociations are competitive with interconversions followed by dissociation.

To date, few studies are conducted to quantify the effects of reduced ammonium (NH4+) and oxidized nitrate (NO3-) on soil CH4 uptake and N2O emission in the subtropical forests. In this study, NH4Cl and NaNO3 fertilizers were applied at three rates: 0, 40 and 120 kg N ha(-1) yr(-1). Soil CH4 and N2O fluxes were determined twice a week using the static chamber technique and gas chromatography. Soil temperature and moisture were simultaneously measured. Soil dissolved N concentration in 0-20 cm depth was measured weekly to examine the regulation to soil CH4 and N2O fluxes. Our results showed that one year of N addition did not affect soil temperature, soil moisture, soil total dissolved N (TDN) and NH4+-N concentrations, but high levels of applied NH4Cl and NaNO3 fertilizers significantly increased soil NO3(-)-N concentration by 124% and 157%, respectively. Nitrogen addition tended to inhibit soil CH4 uptake, but significantly promoted soil N2O emission by 403% to 762%. Furthermore, NH4+-N fertilizer application had a stronger inhibition to soil CH4 uptake and a stronger promotion to soil N2O emission than NO3(-)-N application. Also, both soil CH4 and N2O fluxes were driven by soil temperature and moisture, but soil inorganic N availability was a key integrator of soil CH4 uptake and N2O emission. These results suggest that the subtropical plantation soil sensitively responses to atmospheric N deposition, and inorganic N rather than organic N is the regulator to soil CH4 uptake and N2O emission.

Dissociation of peptide radical ions involves competition between charge-induced and radical-induced reactions that can be preceded by isomerization. The isomeric radical cations of the peptide methyl ester [G˙GR-OMe](+) and [GG˙R-OMe](+) provide very similar collision-induced dissociation (CID) spectra, suggesting that isomerization occurs prior to fragmentation. They undergo characteristic radical-induced bond cleavage of the peptide N-terminal amide bond resulting in the y2(+) ion, and of the arginine side-chain's Cα-Cβ bond giving protonated allylguanidine {[CH2[double bond, length as m-dash]CHCH2NHC(NH2)2](+), m/z 100}. The absence of a y2(+) fragment ion in the CID of the radical cationic tripeptide [ACH3G˙R](+) and of an m/z 100 ion in the spectrum of [G˙ACH3R](+) (where ACH3 is an α-aminoisobutyric acid residue, which cannot form an α-carbon-centered radical through hydrogen atom transfer) establishes the importance of hydrogen atom migration along the peptide backbone prior to specific radical-induced fragmentations. Herein we use density functional theory (DFT) at the B3LYP/6-31++G(d,p) level to evaluate the barriers for interconversion between the α-carbon-centered radicals and for dissociation. The radical cations [G˙GR](+) and [GG˙R](+) have their radicals located on the α-carbon atoms of the peptide backbone and their charge densities largely sequestered on the guanidine groups of the side-chain of arginine residues. This is in contrast to the isomeric radical cations of [GGG]˙(+), in which the charge resides necessarily on the peptide backbone. The lower charge densities on the backbones of [G˙GR](+) and [GG˙R](+) result in greater structural flexibility, decreasing the barrier for interconversion between these α-carbon-centered radicals to 36.2 kcal mol(-1) (cf. 44.7 kcal mol(-1) for [GGG]˙(+)). The total absence of charge, assessed by examining intramolecular hydrogen atom transfers among the three α-carbon centers of the isomeric neutral α-carbon-centered triglycine radicals [GGG-H]˙, leads to an additional but slight reduction in enthalpy, to approximately 34 kcal mol(-1).

A competitive indirect enzyme-linked immunosorbent assay (ELISA) was developed and successfully applied in a dissolved mercury assay. For mercury quantification, factors including the concentration of chelator EDTA, pH, ionic strength, and blocking solution were optimized in this study. Generally the detection limit of the ELISA was standardized as 0.459 μg L−1, with an IC50 value of 13.09 μg L−1 and linear range of 1 to 500 μg L−1; whereas a cross-reactivity study showed that the immunoassay was quite specific for mercury, compared with nine other metal ions. The recovery of the method was evaluated by spiking ultrapure water, tap water and surface water of Taihu Lake with mercury at 1, 10, 50, 100 μg L−1. The precision of the method was evaluated by spiking ultrapure water with mercury at 1, 10, 50, and 100, 500 and 1000 μg L−1. The coefficients of variation of intra- and inter-assay were below 10% in most cases, and recoveries were 94.3–123.5%, 91.9–117.9%, and 92.3–115.6% for ultrapure water, tap water and surface water of Taihu Lake, respectively. Therefore, the developed method is able to determine mercury at levels below the maximum residue limits of water quality standards for drinking water sources of China.