Purification and characterization of a periplasmic laccase produced by Sinorhizobium meliloti

Sinorhizobium meliloti CE52G strain produces a periplasmic laccase that has been purified by a two-step procedure involving heat treatment and immobilized metal affinity chromatography (IMAC). The fraction with laccase activity retained its original activity after 24 h of incubation at pH between 4.0 and 8.0 and after 3 h of incubation at 70 °C, pH 7.2 and supplemented with 1.3 M (NH₄)₂SO₄. It proved to be a homodimeric protein with an apparent molecular mass of 45 kDa each subunit and an isoelectric point of 6.2. CE52G laccase was inhibited by halides (NaF and NaCl), ions (Fe³⁺, Mn²⁺, and Cu²⁺), sulfhydryl organic compounds (β-mercaptoethanol and reduced glutathione), and electron flow inhibitors (NaCN and NaF). Laccase activity was strongly enhanced by (NH₄)₂SO₄, Na₂SO₄, and K₂SO₄. The effects of all these agents, as well as the probability of a partially unfolding polypeptide chain to enhance the interaction between the substrate and the active site, are discussed. CE52G laccase is a pH- and thermo-stable protein with promising biotechnological applications.     

Fen mosses can tolerate some saline conditions found in oil sands process water

Mosses are keystone species in peatlands and are an important part of the vegetation of the pre-mined peatlands. Therefore, mosses should be included in rehabilitation projects following oil sands exploitation in north-western Canada. However, mosses growing in post-mined landscapes must tolerate elevated salinity levels found in oil sands process water (OSPW). Knowledge of salinity tolerance and thresholds for fen mosses is needed to place these mosses in the newly created landscapes. We tested the effects of NaCl and Na₂SO₄ on four fen moss species growing in Petri dishes in growth chambers. We simulated two scenarios: (1) four immersion times (¼, 1, 3 and 7 days) in NaCl (0%, 20%, 60% or 100% of the concentration found in OSPW) mimicking periodic flooding and (2) a permanent saline influence (NaCl or Na₂SO₄ alone or in combination at 0%, 30%, 50% or 70% of the concentrations found in OSPW) mimicking situations of high water tables with different contamination levels. The effects on moss growth were estimated by counting new innovations of Bryum pseudotriquetrum, Campylium stellatum, Sphagnum warnstorfii and Tomenthypnum nitens. All tested mosses tolerated saline levels typically found in post-mined landscapes (up to 500 mg L⁻¹ of NaCl and 400 mg L⁻¹ of Na₂SO₄) for up to 100 days of exposure. Short periods of immersion (up to 7 days independently of salt concentrations) induced the production of innovation in non-Sphagnum species, but S. warnstorfii was more rapidly impacted at higher salt concentrations. Short pulses of salt (from 6 h to 7 days) did not influence the formation of new innovations for C. stellatum and T. nitens. Salt type (NaCl and/or Na₂SO₄) had no effect on moss growth. However, a longer exposure (100 days) with saline water, even at low concentrations, diminished the formation of new innovations for B. pseudotriquetrum and T. nitens. C. stellatum was the least affected by salinity and thus we suggest it is the best species to reintroduce in constructed fens.     

Long-term sulphate and inorganic nitrogen mass balance budgets in European ICP Integrated Monitoring catchments (1990–2012)

Empirical evidence based on integrated environmental monitoring including physical, chemical and biological variables is essential for evaluating the ecosystem benefits of costly emission reduction policies. The international multidisciplinary ICP IM (International Cooperative Programme on Integrated Monitoring of Air Pollution Effects on Ecosystems) programme studies the integrated effects of air pollution and climate change on ecosystems in unmanaged and calibrated forested catchments. We calculated site-specific annual input-output budgets for sulphate (SO₄) and total inorganic nitrogen (TINNO₃-N + NH₄-N) for 17 European ICP IM sites in 1990–2012. Temporal trends for input (deposition) and output (runoff water) fluxes and the net retention/net release of SO₄ and TIN were also analysed. Large differences in the input and output fluxes of SO₄ and TIN reflect important gradients of air pollution effects in Europe, with the highest deposition and runoff water fluxes at IM sites located in southern Scandinavia and in parts of Central and Eastern Europe and the lowest fluxes at more remote sites in northern European regions. A significant decrease in the total (wet + dry) deposition of non-marine SO₄ and bulk deposition of TIN was found at 90% and 65% of the sites, respectively. Output fluxes of non-marine SO₄ in runoff decreased significantly at 65% of the sites, indicating positive effects of the international emission abatement actions in Europe during the last 20 years. Catchments retained SO₄ in the early and mid-1990s, but this shifted towards a net release in the late 1990s, which may be due to the mobilization of legacy S pools accumulated during times of high atmospheric SO₄ deposition. Despite decreased deposition, TIN output fluxes and retention rates showed a mixed response with both decreasing (9 sites) and increasing (8 sites) trend slopes, and trends were rarely significant. In general, TIN was strongly retained in the catchments not affected by natural disturbances. The long-term annual variation in net releases for SO₄ was explained by variations in runoff and SO₄ concentrations in deposition, while a variation in TIN concentrations in runoff was mostly associated with a variation of the TIN retention rate in catchments. The net release of SO₄ from forest soils may delay the recovery from acidification for surface waters and the continued enrichment of nitrogen in catchment soils poses a threat to terrestrial biodiversity and may ultimately lead to a higher TIN runoff through N-saturation. Continued monitoring and further evaluations of mass balance budgets are thus needed.     

Effects of various salts on the germination of three perennial salt marsh species

We studied the germination responses of Arthrocnemum macrostachyum, Juncus acutus and Schoenus nigricans to saline stress caused by different salt types. The germination percentage and mean time to germination data were obtained by incubating seeds for 30 d in 1, 2, 3, 4 or 5% saline solutions of NaCl, MgCl₂, MgSO₄ and Na₂SO₄ at 30/20 °C and with a 12 h photoperiod. A. macrostachyum was the most tolerant species to salinity during the germination (65% in 2% NaCl). S. nigricans showed the lowest germination (none germinated in salt and only 26% in distilled water). J. acutus showed intermediate behaviour between the two above species, its germination being inhibited by high salt concentrations. The sulphates had less inhibitory effect than the equivalent chloride concentrations.     

Adsorption characteristics of sulfur powder by bamboo charcoal to restrain sulfur allergies

Exposures to particulate matter with a diameter of 2.5 μm or less (PM2.5) may influence the risk of birth defects and make you allergic, which causes serious harm to human health. Bamboo charcoal can adsorb harmful substances,that was of benefitto people’s health. In order to figure out the optimal adsorbtion condition and the intrinsic change of bamboo charcoal, five chemicals were adsorbed by bamboo charcoal and were analyzed by FT-IR. The optimal blast time was 80 min of Na₂SO₃, 100 min of Na₂S₂O₈, 20 min of Na₂SO₄, 120 min of Fe₂(SO₄)₃ and 60 min or 100 min of S. FT-IR spectra showed that bamboo charcoal had five characteristic peaks of SS stretch, H₂O stretch, OH stretch, CO stretch or CC stretch, and NO₂ stretch at 3850 cm⁻¹, 3740 cm⁻¹, 3430 cm⁻¹, 1630 cm⁻¹ and 1530 cm⁻¹, respectively. For Na₂SO₃, the peaks at 3850 cm⁻¹, 3740 cm⁻¹, 3430 cm⁻¹, 1630 cm⁻¹ and 1530 cm⁻¹ achieved the maximum at 20 min. For Na₂S₂O₈, the peaks at 3850 cm⁻¹, 3740 cm⁻¹, 3430 cm⁻¹ and 1530 cm⁻¹ achieved the maximum at 40 min. For Na₂SO₄, the peaks at 3850 cm⁻¹, 3740 cm⁻¹ and 1530 cm⁻¹ achieved the maximum at 40 min. For Fe₂(SO₄)₃, the peaks at 3850 cm⁻¹, 3740 cm⁻¹, 1630 cm⁻¹ and 1530 cm⁻¹ achieved the maximum at 120 min. For S, the peaks at 3850 cm⁻¹ and 3740 cm⁻¹ achieved the maximum at 40 min, the peaks at 1630 cm⁻¹ and 1530 cm⁻¹ achieved the maximum at 40 min. It proved that bamboo charcoal could remove sulfur powder from air to restrain sulfur allergies.     

Desulphurization characteristics of bamboo charcoal from sulfur solution

Sulfur powder and sulfur dioxide (SO₂) often floated in air, produced acid rain and algal blooms, and could cause diseases. Bamboo charcoal could have adsorption and filtration properties. In order to figure out the optimal adsorption condition and the intrinsic change of the bamboo charcoal, five chemicals were adsorbed by bamboo charcoal and were analyzed by FT-IR. Fe₂(SO₄)₃’s, Na₂SO₄’s, Na₂S₂O₈’s, S’s, and Na₂SO₃’s optimal adsorption condition was the concentration of 19 g/1000 g and stir time of 20 min, 21 g/1000 g and stir time of 60 min, 7 g/1000 g and stir time of 120 min, 11 g/1000 g and stir time of 120 min, 21 g/1000 g and stir time of 60 min, respectively. FT-IR spectra showed that for FT-IR spectra of Fe₂(SO₄)₃, the transmissivity of the peaks at 3435 cm⁻¹ and 2925 cm⁻¹ achieved the maximum for 60 min and the concentration was 19 g/1000 g, the transmissivity of the peaks at 1630 cm⁻¹, 1060 cm⁻¹ and 660 cm⁻¹ achieved the maximum for 60 min and the concentration was 7 g/1000 g. For FT-IR spectra of Na₂SO₄, the transmissivity of the peaks at 1630 cm⁻¹, 1060 cm⁻¹ and 660 cm⁻¹ achieved the maximum for 20 min and the concentration was 13 g/1000 g. For FT-IR spectra of Na₂S₂O₈, the transmissivity of the peaks at 3435 cm⁻¹, 2925 cm⁻¹, 1630 cm⁻¹ and 1060 cm⁻¹ achieved the maximum for 120 min and the concentration was 19 g/1000 g. For FT-IR spectra of S, the transmissivity of the peaks at 3435 cm⁻¹, 2925 cm⁻¹, 1630 cm⁻¹ and 1060 cm⁻¹ achieved the maximum for 20 min and the concentration was 11 g/1000 g, 17 g/1000 g and 21 g/1000 g. For FT-IR spectra of Na₂SO₃, the transmissivity of the peaks at 3435 cm⁻¹ achieved the maximum for 120 min and the concentration was 5 g/1000 g, the transmissivity of the peaks at 2925 cm⁻¹, 1630 cm⁻¹ and 1060 cm⁻¹ achieved the maximum for 120 min and the concentration was 11 g/1000 g. In these states, the number of the transmissivity of the maximum peaks is the largest.     

Elements of margin of safety,toxicity and action of sodium selenite in a lipopolysaccharide rat model

ProjectBoth septic shock and sodium selenite (Na₂SeO₃) lead to multiple organ failure through oxidation. Na₂SeO₃ has direct oxidant effects above the nutritional level and indirect anti-oxidant properties.In a lipopolysaccharide (LPS) rat model we assessed margin of safety, toxicity and beneficial effect of pentahydrate Na₂SeO₃ (5H₂O·Na₂SeO₃) at oxidant doses.ProcedureIn a three-step study on 204 rats we: (i) observed toxic effects of Na₂SeO₃ injected intraperitoneously (IP) and determined its Minimum Dose Without Toxic effect (MDWT) 0.25–0.35 mg/kg selenium (Se) content; (ii) injected IP LPS at 70% lethal dose (LD) followed, or not, one hour later by IP Na₂SeO₃ at MDWT and (iii) by doses > MDWT. At 48 h, in survivors, we measured plasma creatinine, lactate, aspartate and alanine aminotransferase (AST, ALT), nitric oxide (NO) and Se concentrations.Results(i) Na₂SeO₃ alone did not increase NO and lactate. Encephalopathy appeared at 1 mg Se/kg. Creatinine increased at 1–1.75 mg Se/kg, AST, ALT at 3–4.5 mg Se/kg, and the minimum LD was 3 mg Se/kg. (ii) Mortality after LPS was 37/50 (74%, [62–86%]) vs. 20/30 (67%, [50–84%]) when followed by Na₂SeO₃ at MDWT (p = 0.483) with a decreased in NO (−31%, p = 0.038) a trend for lactate decrease (−19%, p = 0.068) and an increased Se in plasma of survivals. (iii) All rats died at doses ≥0.6 mg/kg (p < 0.001).ConclusionMechanisms of LPS and Na₂SeO₃ toxicity differ (i.e. NO, lactate). In septic shock 5H₂O·Na₂SeO₃ toxicity increased, margin of safety decrease, but IP administration of dose considered as oxidant of 5H₂O·Na₂SeO₃ showed beneficial effects.     

Differential tolerance to combined salinity and O2 deficiency in the halophytic grasses Puccinellia ciliata and Thinopyrum ponticum: The importance of K+ retention in roots

Saline environments of terrestrial halophytes are often prone to waterlogging, yet the effects on halophytes of combined salinity and waterlogging have rarely been studied. Either salinity or hypoxia (low O₂) alone can interfere with K⁺ homeostasis, therefore the combination of salinity or hypoxia is expected to impact significantly on K⁺ retention in roots. We studied mechanisms of tolerance to the interaction of salinity with hypoxia in Puccinellia ciliata and Thinopyrum ponticum, halophytic grasses that differ in waterlogging tolerance. Plants were exposed to aerated and stagnant saline (250 mM NaCl) treatments with low (0.25 mM) and high (4 mM) K⁺ levels; growth, net ion fluxes and tissue ion concentrations were determined. P. ciliata was more tolerant than T. ponticum to stagnant-saline treatment, producing twice the biomass of adventitious roots, which accumulated high levels of Na⁺, and had lower shoot Na⁺. After 24 h of saline hypoxic treatment, MIFE measurements revealed a net uptake of K⁺ (∼40 nmol m⁻² s⁻¹) for P. ciliata, but a net loss of K⁺ (∼20 nmol m⁻² s⁻¹) for the more waterlogging sensitive T. ponticum. NaCl alone induced K⁺ efflux from roots of both species, with channel blocker tests implicating GORK-like channels. P. ciliata had constitutively a more negative root cell membrane potential than T. ponticum (−150 versus −115 mV). Tolerance to salinity and hypoxia in P. ciliata is related to increased production of adventitious roots, regulation of shoot K⁺/Na⁺, and a superior ability to maintain negative membrane potential in root cells, resulting in greater retention of K⁺.     

Synthesis of 1-(methanesulfonyl- and aminosulfonylphenyl)acetylenes that possess a 2-(N-difluoromethyl-1,2-dihydropyridin-2-one) pharmacophore: Evaluation as dual inhibitors of cyclooxygenases and 5-lipoxygenase with anti-inflammatory activity

A hitherto unknown class of linear acetylene regioisomers were designed such that a SO₂Me or SO₂NH₂ group was located at the ortho-, meta- or para-position of the acetylene C-1 phenyl ring, and a N-difluoromethyl-1,2-dihydropyridin-2-one moiety was attached via its C-5 position to the C-2 position on an acetylene template (scaffold). All three SO₂Me regioisomers, and the 4-SO₂NH₂ analog, were potent inhibitors of 5-lipoxygenase (5-LOX IC₅₀ = 3.2–3.5 μM range) relative to the reference drug caffeic acid (IC₅₀ = 4.0 μM). The SO₂Me regioisomers exhibited weak cyclooxygenease-1 (COX-1) and -2 (COX-2) inhibitory activity with a modest COX-2 selectivity index. The most potent 3-SO₂Me, 4-SO₂Me and 4-SO₂NH₂ compounds, with respective ED₅₀ values of 66.1, 68.5 and 86.5 mg/kg po, exhibited comparable oral anti-inflammatory (AI) activity to that of the reference drug ibuprofen (ED₅₀ = 67.4 mg/kg po). The N-difluoromethyl-1,2-dihydropyridin-2-one moiety provides a novel pharmacophore for the design of cyclic hydroxamic mimetics capable of inhibiting 5-LOX for exploitation in the development of 5-LOX inhibitory AI drugs.     

Characteristics of activated carbon remove sulfur particles against smog

Sulfur particles, which could cause diseases, were the main powder of smog. And activated carbon had the very adsorption characteristics. Therefore, five sulfur particles were adsorbed by activated carbon and were analyzed by FT-IR. The optimal adsorption time were 120 min of Na₂SO₃, 120 min of Na₂S₂O₈, 120 min of Na₂SO₄, 120 min of Fe₂(SO₄)₃ and 120 min of S. FT-IR spectra showed that activated carbon had the eight characteristic absorption of SS stretch, H₂O stretch, OH stretch, CH stretch, conjugated CO stretch or CC stretch, CH₂ bend, CO stretch and acetylenic CH bend vibrations at 3850 cm^–1, 3740 cm^–1, 3430 cm^–1, 2920 cm^–1, 1630 cm^–1, 1390 cm^–1, 1110 cm^–1 and 600 cm^–1, respectively. For Na₂SO₃, the peaks at 2920 cm^–1, 1630 cm^–1, 1390 cm^–1 and 1110 cm^–1 achieved the maximum at 20 min. For Na₂S₂O₈, the peaks at 3850 cm^–1, 3740 cm^–1 and 2920 cm^–1 achieved the maximum at 60 min. The peaks at 1390 cm^–1, 1110 cm^–1 and 600 cm^–1 achieved the maximum at 40 min. For Na₂SO₄, the peaks at 3430 cm^–1, 2920 cm^–1, 1630 cm^–1, 1390 cm^–1, 1110 cm^–1 and 600 cm^–1 achieved the maximum at 60 min. For Fe₂(SO₄)₃, the peaks at 1390 cm^–1, 1110 cm^–1 and 600 cm^–1 achieved the maximum at 20 min. For S, the peaks at 1630 cm^–1, 1390 cm^–1 and 600 cm^–1 achieved the maximum at 120 min. It provided that activated carbon could remove sulfur particles from smog air to restrain many anaphylactic diseases.     

Sodium orthovanadate associated with pharmacological doses of ascorbate causes an increased generation of ROS in tumor cells that inhibits proliferation and triggers apoptosis

Pharmacological doses of ascorbate were evaluated for its ability to potentiate the toxicity of sodium orthovanadate (Na₃VO₄) in tumor cells. Cytotoxicity, inhibition of cell proliferation, generation of ROS and DNA fragmentation were assessed in T24 cells. Na₃VO₄ was cytotoxic against T24 cells (EC₅₀ = 5.8 μM at 24 h), but in the presence of ascorbate (100 μM) the EC₅₀ fell to 3.3 μM. Na₃VO₄ plus ascorbate caused a strong inhibition of cell proliferation (up to 20%) and increased the generation of ROS (4-fold). Na₃VO₄ did not directly cleave plasmid DNA, at this aspect no synergism was found occurring between Na₃VO₄ and ascorbate once the resulting action of the combination was no greater than that of both substances administered separately. Cells from Ehrlich ascites carcinoma-bearing mice were used to determine the activity of antioxidant enzymes, the extent of the oxidative damage and the type of cell death. Na₃VO₄ alone, or combined with ascorbate, increased catalase activity, but only Na₃VO₄ plus ascorbate increased superoxide dismutase activity (up to 4-fold). Oxidative damage on proteins and lipids was higher due to the treatment done with Na₃VO₄ plus ascorbate (2–3-fold). Ascorbate potentiated apoptosis in tumor cells from mice treated with Na₃VO₄. The results indicate that pharmacological doses of ascorbate enhance the generation of ROS induced by Na₃VO₄ in tumor cells causing inhibition of proliferation and apoptosis. Apoptosis induced by orthovanadate and ascorbate is closer related to inhibition on Bcl-xL and activation of Bax. Our data apparently rule out a mechanism of cell demise p53-dependent or related to Cdk2 impairment.     

Novel fibrinolytic enzymes from Virgibacillus halodenitrificans SK1-3-7 isolated from fish sauce fermentation

Virgibacillus sp. SK1-3-7 exhibited the highest fibrinolytic activity among 25 bacterial isolates obtained from fish sauce fermentation. Results of 16S rRNA gene sequence analysis showed 99% homology to Virgibacillus halodenitrificans ATCC 49067. It was, therefore, identified as V. halodenitrificans SK1-3-7. Fibrinolytic enzymes from V. halodenitrificans SK1-3-7 were partially purified using ammonium sulfate fractionation, hydrophobic and ion-exchange chromatographies. The enzymes with molecular weight of 20- and 36-kDa showed fibrinolytic activity on a fibrin zymogram. The enzymes were stable between pH 4 and 10 and below 60 °C. The enzymes were activated by 20 mM CaCl₂ and 0.15 M NaCl. The activity increased with CaCl₂ up to 100 mM and increased with NaCl concentration up to 2 M. In addition, the residual fibrinolytic activity of 61% was found at 4 M NaCl. The enzymes were completely inhibited by phenylmethanesulfonyl fluoride (PMSF) and preferably hydrolyzed Suc-Ala-Ala-Pro-Phe-pNA, suggesting a subtilisin-like serine proteinase. V. halodenitrificans SK1-3-7 enzymes hydrolyzed fibrin to a greater extent than did plasmin. In addition, the enzymes were resistant to pepsin and trypsin digestion. The de novo peptide homology analysis of a 20- and 36-kDa proteinase revealed no matches to bacilli serine proteinases, suggesting that both were novel fibrinolytic enzymes.     

Production of hemicellulosic sugars and glucose from residual corrugated cardboard

Corrugated cardboard samples were subjected to two-step saccharification. A first prehydrolysis stage was carried out to solubilise the hemicellulosic fraction as hemicellulosic sugars, and the solid phase from prehydrolysis was used as a substrate for the enzymic hydrolysis of cellulose. The prehydrolysis step was carried out for 0–180 min in media containing 1–3 wt.% of H₂SO₄ and the fraction of solid recovered after treatments and the compositions of solid and liquid phases from treatments were measured. The susceptibility of prehydrolysed solids towards the enzymic hydrolysis was assessed in further experiments. Under selected prehydrolysis conditions (3% H₂SO₄, 180 min), 78.2% of initial hemicelluloses was saccharified, leading to liquors containing up to 10 g hemicellulosic sugars/l and 9.2 g glucose/l. The corresponding solid phase, enriched in cellulose, showed good susceptibility towards enzymatic hydrolysis, leading to solutions containing up to 17.9 g glucose/l (conversion yield=63.6%) and a glucose/total sugar ratio of 0.93 g/g. Mathematical models assessing the effects of the operational conditions on both the prehydrolysis stage and the susceptibility of substrates towards enzymic hydrolysis have been developed.     

Activation of nylon net and its application to a biosensor for determination of glucose in human serum

A glucose biosensor using a glucose oxidase (GO_x)-immobilized nylon net with glutaraldehyde as cross-linking reagent and an oxygen (O₂) electrode for the determination of glucose has been fabricated. The detection scheme was based on the utilization of dissolved O₂ in oxidation of glucose by the membrane bound GO_x. Crucial factors including O-alkylation temperature, reaction times of nylon net with dimethyl sulfate, l-lysine, and glutaraldehyde, and enzyme loading were examined to determine the optimal enzyme immobilization conditions for the best sensitivity of the developed glucose biosensor. In addition, the effects of pH and concentration of phosphate buffer on the response of the biosensor were studied. The glucose biosensor had a linear range of 18 μM to 1.10 mM with the detection limit of 9.0 μM (S/N = 3) and response time of 80 s. The biosensor exhibited both good operational stability with over 200 measurements and long-term storage stability. The results from this biosensor compared well with those of a commercial glucose assay kit in analyzing human serum glucose samples.     

The combined effects of gibberellic acid and salinity on some antioxidant enzyme activities,plant growth parameters and nutritional status in maize plants

The combined effects of salt stress and gibberellic acid (GA₃) on plant growth and nutritional status of maize (Zea mays L. cv., DK 647 F1) were studied in a pot experiment. Treatments were (1) control (C): nutrient solution alone, (2) salt stress (S): 100 mM NaCl, (3) S + GA1: 100 mM NaCl and 50 ppm GA₃ and (4) S + GA2: 100 mM NaCl and 100 ppm GA₃. Salt stress (S) was found to reduce the total dry matter, chlorophyll content, relative water content (RWC), but to increase proline accumulation, superoxide dismutase (SOD; EC 1.15.1.1), peroxidase (POD; EC 1.11.1.7) and polyphenol oxidase (PPO; 1.10.3.1) enzyme activities and electrolyte leakage. GA₃ treatments overcame to variable extents the adverse effects of NaCl stress on the above physiological parameters. GA₃ treatments reduced the activities of enzyme in the salt-stressed plants. Salt stress reduced some macro and micronutrient concentrations but exogenous application of GA₃ increased these to levels of control treatment. Foliar application of GA₃ counteracted some of the adverse effects of NaCl salinity with the accumulation of proline which maintained membrane permeability and increased macro and micronutrient levels.     

Soil nitrate accumulation explains the nonlinear responses of soil CO2 and CH4 fluxes to nitrogen addition in a temperate needle-broadleaved mixed forest

The responses of soil-atmosphere carbon (C) exchange fluxes to growing atmospheric nitrogen (N) deposition are controversial, leading to large uncertainty in the estimated C sink of global forest ecosystems experiencing substantial N inputs. However, it is challenging to quantify critical load of N input for the alteration of the soil C fluxes, and what factors controlled the changes in soil CO₂ and CH₄ fluxes under N enrichment. Nine levels of urea addition experiment (0, 10, 20, 40, 60, 80, 100, 120, 140 kg N ha⁻¹ yr⁻¹) were conducted in the needle-broadleaved mixed forest in Changbai Mountain, Northeast China. Soil CO₂ and CH₄ fluxes were monitored weekly using the static chamber and gas chromatograph technique. Environmental variables (soil temperature and moisture in the 0–10 cm depth) and dissolved N (NH₄⁺-N, NO₃⁻-N, total dissolved N (TDN), and dissolved organic N (DON)) in the organic layer and the 0–10 cm mineral soil layer were simultaneously measured. High rates of N addition (≥60 kg N ha⁻¹ yr⁻¹) significantly increased soil NO₃⁻-N contents in the organic layer and the mineral layer by 120%-180% and 56.4%-84.6%, respectively. However, N application did not lead to a significant accumulation of soil NH₄⁺-N contents in the two soil layers except for a few treatments. N addition at a low rate of 10 kg N ha⁻¹ yr⁻¹ significantly stimulated, whereas high rate of N addition (140 kg N ha⁻¹ yr⁻¹) significantly inhibited soil CO₂ emission and CH₄ uptake. Significant negative relationships were observed between changes in soil CO₂ emission and CH₄ uptake and changes in soil NO₃⁻-N and moisture contents under N enrichment. These results suggest that soil nitrification and NO₃⁻-N accumulation could be important regulators of soil CO₂ emission and CH₄ uptake in the temperate needle-broadleaved mixed forest. The nonlinear responses to exogenous N inputs and the critical level of N in terms of soil C fluxes should be considered in the ecological process models and ecosystem management.     

Alleviating the adverse effects of NaCl stress in maize seedlings by pretreating seeds with salicylic acid and 24-epibrassinolide

The effects of bio-regulators salicylic acid (SA) and 24-epibrassinolide (EBL) as seed soaking treatment on the growth traits, content of photosynthetic pigments, proline, relative water content (RWC), electrolyte leakage percent (EC%), antioxidative enzymes and leaf anatomy of Zea mays L. seedlings grown under 60 or 120 mM NaCl saline stress were studied. A greenhouse experiment was performed in a completely randomized design with nine treatments [control (treated with tap water); 60 mM NaCl; 120 mM NaCl; 10^− 4 M SA; 60 mM NaCl + 10^− 4 M SA; 120 mM NaCl + 10^− 4 M SA; 10 μM EBL; 60 mM NaCl + 10 μMEBL or 120 mM NaCl + 10 μM EBL] each with four replicates. The results indicated that NaCl stress significantly reduced plant growth traits, leaf photosynthetic pigment, soluble sugars, RWC%, and activities of catalase (CAT), peroxidase (POX) as well as leaf anatomy. However, the application of SA or EBL mitigated the toxic effects of NaCl stress on maize seedlings and considerably improved growth traits, photosynthetic pigments, proline, RWC%, CAT and POX enzyme activities as well as leaf anatomy. This study highlights the potential ameliorative effects of SA or EBL in mitigating the phytotoxicity of NaCl stress in seeds and growing seedlings.     

Adsorption characteristics of sulfur solution by acticarbon against drinking-water toxicosis

Sulfur and ammonia nitrogen are rich nutrient pollutants, after entering water can cause algal blooms, cause eutrophication of water body, the spread of them will not only pollute the environment, destroy the ecological balance, but also harm human health through food chain channels, especially drinking-water toxicosis. Acticarbon can adsorb harmful substances, it was beneficial for people’s health. In order to figure out the optimal adsorption condition and the intrinsic change of acticarbon, five chemicals were adsorbed by acticarbon and analyzed by FT-IR. The optimal adsorption condition of Fe₂(SO₄)₃, Na₂SO₄, Na₂S₂O₈, S and Na₂SO₃ was 9 g/1000 g at 80 min, 21 g/1000 g at 20 min, 15g/1000 g at 20 min, 21 g/1000 g at 60 min and 21 g/1000 g at 100 min, respectively. FT-IR spectra showed that acticarbon had eight characteristic peaks, such as S-S stretch, H₂O stretch, OH stretch, CH stretch, CO or CC stretch, CH₂ bend, CH were at 3850 cm⁻¹, 3740 cm⁻¹, 3435 cm⁻¹, 2925 cm⁻¹, 1630 cm⁻¹, 1390 cm⁻¹, 1115 cm⁻¹, 600 cm⁻¹, respectively. For FT-IR spectra of Fe₂(SO₄)₃, the peaks at 3850 cm⁻¹, 3740 cm⁻¹, 2925 cm⁻¹ achieved the maximum with 9 g/1000 g at 20 min. For Na₂SO₄, the peaks at 2925 cm⁻¹, 1630 cm⁻¹, 1390 cm⁻¹, 1115 cm⁻¹, 600 cm⁻¹ achieved the maximum with 21 g/1000 g at 120 min. For ones of Na₂S₂O₈, the peaks at 3850 cm⁻¹, 3740 cm⁻¹, 1390 cm⁻¹, 1115 cm⁻¹, 600 cm⁻¹, achieved the maximum with 2 g/1000 g at 80 min. For ones of S, the peaks at 3850 cm⁻¹, 3740 cm⁻¹, 2925 cm⁻¹ achieved the maximum with 19 g/1000 g at 100 min, the peaks at 1390 cm⁻¹, 1115 cm⁻¹, 600 cm⁻¹ achieved the maximum with 19 g/1000 g at 20 min. For FT-IR spectra of Na₂SO₃, the peaks at 1630 cm⁻¹, 1390 cm⁻¹, 1115 cm⁻¹, 600 cm⁻¹ achieved the maximum with 2 g/1000 g at 100 min. It provided that acticarbon could adsorb and desulphurize from sulfur solution against drinking-water toxicosis.     

1,4-Diaryl-substituted triazoles as cyclooxygenase-2 inhibitors: Synthesis,biological evaluation and molecular modeling studies

A novel group of 1,4-diaryl-substituted triazoles was designed and synthesized by introducing the cyclooxygenase-2 (COX-2) pharmacophore SO₂NH₂ attached to one aryl ring and various substituents (H, F, Cl, CH₃ or OCH₃) attached to the other aryl ring. The effects of size and flexibility of the compounds upon COX-1/COX-2 inhibitory potency and selectivity was studied by increasing the size of an alkyl linker chain [(–CH₂)_n, where n = 0, 1, 2]. In vitro COX-1/COX-2 inhibition studies showed that all compounds (14–18, 21–25 and 28–32) are more potent inhibitors of COX-2 isozyme (IC₅₀ = 0.17–28.0 μM range) compared to COX-1 isozyme (IC₅₀ = 21.0 to >100 μM range). Within the group of 1,4 diaryl-substituted triazoles, 4-{2-[4-(4-chloro-phenyl)-[1,2,3]triazol-1-yl]-ethyl}-benzenesulfonamide (compound 30) displayed highest COX-2 inhibitory potency and selectivity (COX-1: IC₅₀ = >100 μM, COX-2: IC₅₀ = 0.17 μM, SI >588). Molecular docking studies using the catalytic site of COX-1 and COX-2, respectively, provided complementary theoretical support for the obtained experimental biological structure–activity relationship data. Results of molecular docking studies revealed that COX-2 pharmacophore SO₂NH₂ in compound 30 is positioned in the secondary pocket of COX-2 active site; with the nitrogen atom of the SO₂NH₂ group being hydrogen bonded to Q192 (N?OC = 2.85 Å), and one of the oxygen atoms of SO₂NH₂ group forming a hydrogen bond to H90 (SO?N = 2.38 Å).     

The effects of calcium sulphate on growth,membrane stability and nutrient uptake of tomato plants grown under salt stress

A pot experiment was carried out with tomato (Lycopersicon esculentum Mill.) cv. “Target F1” in a mixture of peat, perlite, and sand (1:1:1) to investigate the effects of supplementary calcium sulphate on plants grown at high NaCl concentration (75 mM). The treatments were: (i) control (C), nutrient solution alone; (ii) salt treatment (C + S), 75 mM NaCl; (iii) salt plus calcium treatment 1 (C + S + Ca1), 75 mM NaCl plus additional mixture of 2.5 mM CaSO₄ in nutrient solution; (iv) salt plus calcium treatment 2 (C + S + Ca2), 75 mM NaCl plus additional mixture of 5 mM CaSO₄ in nutrient solution. The plants grown under salt stress produced low dry matter, fruit weight, and relative water content than those grown in standard nutrient solution. Supplemental calcium sulphate added to nutrient solution containing salt significantly improved growth and physiological variables affected by salt stress (e.g. plant growth, fruit yield, and membrane permeability) and also increased leaf K⁺, Ca²⁺, and N in tomato plants. The effects of supplemental CaSO₄ in maintaining membrane permeability, increasing concentrations of Ca²⁺, N, and K⁺ and reducing concentration of Na⁺ (because of cation competition in root zone) in leaves could offer an economical and simple solution to tomato crop production problems caused by high salinity.