Proteomic analysis of the effects of tumor necrosis factor-α on endothelial cells

AIM: To investigate the affected proteins by tumor necrosis factor (TNF)-α in endothelial cells, and further explore the potential molecular mechanism of TNF-α on endothelial cells. METHODS: Nitric oxide (NO) production in the cultured human umbilical vein endothelial cells (HUVECs) was measured by a NO assay kit. Proteomic alterations were analyzed using two-dimensional electrophoresis, and peptide mass fingerprinting with matrix-assisted laser desorption/ionization-time of flight mass spectrometry. RESULTS: NO production in HUVECs decreased significantly after TNF-α treatement. Proteomics analysis showed 21 protein spots were changed including 9 spots that were increased and 11 spots that were decreased after TNF-α stimulation, and 1 spot was only detected in TNF-α activated cell gels. CONCLUSIONS: Decreased the expression of ecNOS by TNF-α might result in decreased NO production. Up-regulated MAP/ERK kinase 3 expression might imply that TNF-α activates the expression of adhesion molecules. Cytoskeletal protein actin is also involved in TNF-α injuried HUVECs. Proteomic analysis can find some clues for identifying new potential target of TNF-α.     

不同锌源对断奶仔猪免疫和抗氧化作用的影响

选用 1 4 4头断奶仔猪 ,随机分成 4组 :一组为对照 ,饲喂添加氧化锌 1 0 0 mg/kg的日粮 ,另 3组分别饲喂添加氧化锌 30 0 0 m g/kg、蛋氨酸锌 1 0 0 m g/kg和纳米氧化锌 30 0 m g/kg的日粮 ,进行为期 5 2 d的饲养试验 ,以研究不同锌源对断奶仔猪消化和免疫功能的影响。结果表明 :高剂量氧化锌可提高血清中 Ig A的含量 5 .0 7%(P<0 .0 5 ) ,高剂量氧化锌和纳米氧化锌可使血清中免疫球蛋白 Ig M含量分别提高 2 .97%(P<0 .0 5 )和 5 .6 7%(P<0 .0 1 ) ;蛋氨酸锌可使血清中SOD活性比对照组提高 1 .36倍 (P<0 .0 1 ) ,高剂量氧化锌和蛋氨酸锌可分别使肝脏中 SOD活性提高 2 8.2 0 %(P<0 .0 5 )和 2 9.33%(P<0 .0 5 ) ;高剂量氧化锌、蛋氨酸锌和纳米氧化锌均可提高肝脏组织中金属硫蛋白 (MT)含量     

NO介导的Ca2+信号对渗透胁迫下紫花苜蓿幼苗光合特征及抗性的影响

以紫花苜蓿幼苗为材料,用聚乙二醇(PEG-6000)作为渗透介质人工模拟干旱条件,外源喷施NO供体硝普钠(SNP)、钙信号试剂CaCl₂、NO抑制剂亚甲基蓝(MB)和Ca²⁺通道阻断剂LaCl₃,对紫花苜蓿幼苗光合特征、抗氧化酶活性及过氧化物酶(POD)同工酶图谱进行研究,探讨了渗透胁迫下NO介导的Ca²⁺信号对紫花苜蓿幼苗光合作用及抗氧化能力的影响。结果表明:在渗透胁迫条件下,施加SNP、CaCl₂均能够有效缓解叶片叶绿素a、类胡萝卜素及总叶绿素含量降低,提高叶片净光合速率(P_n)、气孔导度(G_s)及气孔限制值(L_s),而对胞间CO₂浓度(C_i)没有缓解作用。SNP、CaCl₂及SNP+CaCl₂处理提高了幼苗叶片中抗氧化酶活性和脯氨酸含量,降低了丙二醛(MDA)含量。其中共处理时效果最为显著,第4天 SOD、POD、CAT活性较PEG处理升高了39.29%、30.41%和56.24%,脯氨酸含量增加了45.59%,MDA含量降低了45.59%。POD同工酶图谱在第4天时酶谱带数最多,POD活性最强,且SNP+CaCl₂共处理下出现新酶带。而添加外源NO的同时添加Ca²⁺通道阻断剂LaCl₃,紫花苜蓿幼苗光合速率、抗氧化酶活性及脯氨酸含量均降低,丙二醛含量增加,添加Ca²⁺信号的同时施加NO抑制剂MB也具有相同的作用,说明Ca²⁺信号参与NO信号转导过程并相互作用共同调节渗透胁迫下紫花苜蓿幼苗的生理应答响应。     

Effects of zinc oxide nanoparticles on the egg quality,immune response,zinc retention,and blood parameters of laying hens in the late phase of production

The objective of this study was to evaluate the effects of dietary supplementation with zinc oxide nanoparticles (ZnO‐NPs) on the performance, egg quality, Zn retention, immunity responses, superoxide dismutase activity (SOD), egg malondialdehyde (MDA) content, and serum parameters in laying hens in the late phase of production. A total of 288 laying hens at 64 weeks of age were randomly assigned to 4 treatments with 6 replicates, and 12 birds within each group. Experimental diets included a corn‐soybean meal‐based diet (without Zn supplementation) and a basal diet supplemented with 80 mg/kg of Zn‐oxide, ZnO‐NPs, and Zn‐methionine. The results indicated that egg production and egg mass were significantly higher in the Zn‐methionine and ZnO‐NPs groups (p < .05). Also, eggshell thickness and shell strength increased in the ZnO‐NPs group as compared with the other groups (p < .05). Moreover, Zn supplementation decreased egg loss (p < .05). There were significant differences among treatments in Zn deposition in tibiotarsus, liver, pancreas, eggs, and excreta (p < .01). Antibody titre, heterophil (%(, and phytohemagglutinin (PHA) were significantly higher in birds fed with Zn‐supplemented diets (p < .05). In treatments supplemented with ZnO‐NPs and Zn‐methionine, the SOD activity in the liver, pancreas, and plasma was greater as compared with the other treatments (p < .05). The MDA content in eggs was significantly reduced in groups supplemented with Zn (p < .01). Moreover, dietary Zn supplementation significantly affected serum total protein, albumin, glucose, alkaline phosphatase activity, carbonic anhydrase activity, and Zn level (p < .05). In conclusion, this study demonstrated that dietary supplementation with ZnO‐NPs can improve the performance of laying hens. Therefore, ZnO‐NPs can enhance zinc absorption in the intestine of aged layers and can be a more suitable source of zinc than regular Zn‐oxide in diets.     

Nitrous oxide emissions from dairy farm effluent applied to a New Zealand pasture soil

A field experiment on permanent ryegrass–white clover pasture at AgResearch's Ruakura dairy farm near Hamilton, New Zealand quantified nitrous oxide (N₂O) emissions from different types of dairy effluent applied to soil at three seasons and evaluated the potential of dicyandiamide (DCD) (a nitrification inhibitor) to decrease gaseous N₂O emissions. Fresh or stored manure and farm dairy effluent (FDE; from dairy shed washings), with or without DCD (10 kg/ha), were applied at approximately 100 kg N/ha to plots on a well‐drained soil on volcanic parent material. A field chamber technique was used to measure N₂O emissions. Application of manure or FDE, both in fresh and stored forms, to pasture generally increased N₂O emissions. Overall N₂O emission factors (EF) varied between 0.01% and 1.87%, depending on application season and effluent type. EFs in spring and autumn were greater than those in summer (P < 0.05). Among the effluents, N₂O EFs were largest from fresh FDE (1.65%) during the spring measurement period, stored manure (1.87%) during the autumn and stored FDE (0.25%) during the summer. DCD was effective in decreasing N₂O EFs from fresh FDE, fresh manure, stored FDE and stored manure by 40–80%, 69–76%, 24–84% and 60–70%, respectively. DCD reduced N₂O emissions during the spring and autumn seasons more effectively than in the summer season.     

维生素A调节动物一氧化氮的生成及其相关机制

动物机体由于旺盛的新陈代谢或在受到外界有害因素刺激的情况下会产生大量一氧化氮(NO),进而损伤细胞中的蛋白质、类脂膜和DNA,引发细胞炎症并阻断细胞信号通路,使机体产生氧化应激。维生素A可以有效地调控NO的生成,提高机体抗氧化水平并清除自由基,预防细胞炎症及氧化应激的发生。本文主要综述了维生素A对动物NO生成的调节作用及其相关机理的研究进展,对今后深入研究其调节机理、科学补充维生素A及提高机体的抗氧化功能具有一定参考价值。     

A review of soil NO transformation: Associated processes and possible physiological significance on organisms

NO emissions from soils and ecosystems are of outstanding importance for atmospheric chemistry. Here we review the current knowledge on processes involved in the formation and consumption of NO in soils, the importance of NO for the physiological functioning of different organisms, and for inter- and intra-species signaling and competition, e.g. in the rooting zone between microbes and plants. We also show that prokaryotes and eukaryotes are able to produce NO by multiple pathways and that unspecific enzymo-oxidative mechanisms of NO production are likely to occur in soils. Nitric oxide production in soils is not only linked to NO production by nitrifying and denitrifying microorganisms, but also linked to extracellular enzymes from a wide range of microorganisms.Further investigations are needed to clarify molecular mechanisms of NO production and consumption, its controlling factors, and the significance of NO as a regulator for microbial, animal and plant processes. Such process understanding is required to elucidate the importance of soils as sources (and sinks) for atmospheric NO.     

Redox halopriming: A Promising Strategy for Inducing Salt Tolerance in Bread Wheat

New strategies to enhance growth and productivity of food crops in saline soils represent important research priorities. This study has investigated the role of certain priming techniques to induce salt tolerance of bread wheat. Wheat grains were soaked in 0.2 mm sodium nitroprusside as nitric oxide donor (redox priming), diluted sea water (halopriming) and the combination of both (redox halopriming). Grains were also soaked in distilled water (hydropriming); in addition, untreated grains were taken as control. Our results indicated that priming treatments significantly improved all growth traits and increased leaf pigments concentration as compared to the control. Priming treatments markedly enhanced membrane stability index, proline, total soluble sugars and K⁺ concentration with simultaneous decrease in the concentration of Na⁺ and malondialdehyde (MDA). Furthermore, yield and yield‐related traits such as plant height, spike length, total number of tillers, 1000‐grain weight, straw and grain yield considerably affected by priming treatments. Moreover, the grain yield of both genotypes was positively affected by redox halopriming treatment. However, the extent of enhancement was more prominent in Gemmiza‐9 (salt sensitive) than that in Sakha‐93 (salt‐tolerant). Overall, this study clearly indicated that redox halopriming treatment is a promising and handy technique to induce salinity tolerance of wheat genotypes.     

Fungal and bacterial N2O production regulated by soil amendments of simple and complex substrates

Fungal N₂O production results from a respiratory denitrification that reduces NO₃⁻/NO₂⁻ in response to the oxidation of an electron donor, often organic C. Despite similar heterotrophic nature, fungal denitrifiers may differ from bacterial ones in exploiting diverse resources. We hypothesized that complex C compounds and substances could favor the growth of fungi over bacteria, and thereby leading to fungal dominance for soil N₂O emissions. Effects of substrate quality on fungal and bacterial N₂O production were, therefore, examined in a 44-d incubation after soils were amended with four different substrates, i.e., glucose, cellulose, winter pea, and switchgrass at 2 mg C g⁻¹ soil. During periodic measurements of soil N₂O fluxes at 80% soil water-filled pore space and with the supply of KNO₃, substrate treatments were further subjected to four antibiotic treatments, i.e., no antibiotics or soil addition of streptomycin, cycloheximide or both so that fungal and bacterial N₂O production could be separated. Up to d 8 when antibiotic inhibition on substrate-induced microbial activity and/or growth was still detectable, bacterial N₂O production was generally greater in glucose- than in cellulose-amended soils and also in winter pea- than in switchgrass-amended soils. In contrast, fungal N₂O production was more enhanced in soils amended with cellulose than with glucose. Therefore, fungal-to-bacterial contribution ratios were greater in complex than in simple C substrates. These ratios were positively correlated with fungal-to-bacterial activity ratios, i.e., CO₂ production ratios, suggesting that substrate-associated fungal or bacterial preferential activity and/or growth might be the cause. Considering substrate depletion over time and thereby becoming limited for microbial N₂O production, measurements of soil N₂O fluxes were also carried out with additional supply of glucose, irrespective of different substrate treatments. This measurement condition might lead to potentially high rates of fungal and bacterial N₂O production. As expected, bacterial N₂O production was greater with added glucose than with added cellulose on d 4 and d 8. However, this pattern was broken on d 28, with bacterial N₂O production lower with added glucose than with added cellulose. In contrast, plant residue impacts on soil N₂O fluxes were consistent over 44-d, with greater bacterial contribution, lower fungal contribution, and thus lower fungal-to-bacterial contribution ratios in winter pea- than in switchgrass-amended soils. Real-time PCR analysis also demonstrated that the ratios of 16S rDNA to ITS and the copy numbers of bacterial denitrifying genes were greater in winter pea- than in switchgrass-amended soils. Despite some inconsistency found on the impacts of cellulose versus glucose on fungal and bacterial leading roles for N₂O production, the results generally supported the working hypothesis that complex substrates promoted fungal dominance for soil N₂O emissions.