Comparison in copper accumulation and physiological responses of Gracilaria lemaneiformis and G. lichenoides (Rhodophyceae)

Heavy metal pollution has become a worldwide problem in aquaculture. We studied copper (Cu2+ ) accumulation and physiological responses of two red algae Gracilaria lemaneiformis and Gracilaria lichenoides from China under Cu2+ exposure of 0-500 μg/L in concentration. Compared with G. lemaneiformis, G. lichenoides was more capable in accumulating Cu2+ , specifically, more Cu2+ on extracellular side (cell wall) than on intracellular side (cytoplasm) and in cell organelles (especially chloroplast, cell nucleus, mitochondria, and ribosome). In addition, G. lichenoides contained more insoluble polysaccharide in cell wall, which might promote the extracellular Cu2+ -binding as an efficient barrier against metal toxicity. Conversely, G. lemaneiformis was more vulnerable than G. lichenoides to Cu 2+ toxin for decreases in growth, pigment (chlorophyll a, chlorophyll b, phycobiliprotein, and β-carotene) content, and photosynthetic activity. Moreover, more serious oxidative damages in G. lemaneiformis than in G. lichenoides, in accumulation of reactive oxidative species and malondialdehyde, and in electrolyte leakage, because of lower antioxidant enzyme (superoxide dismutase and glutathione reductase) activities. Therefore, G. lichenoides was less susceptible to Cu2+ stress than G. lemaneiformis.     

Optimization of conditions for tetraspore release and assessment of photosynthetic activities for different generation branches of Gracilaria lemaneiformis Bory

Gracilaria lemaneiformis Bory is an economically important alga that is primarily used for agar production. Although tetraspores are ideal seeds for the cultivation of G. lemaneiformis, the most popular culture method is currently based on vegetative fragments, which is labor-intensive and time-consuming. In this study, we optimized the conditions for tetraspore release and evaluated the photosynthetic activities of different colonies formed from the branches of G. lemaneiformis using a PAM (pulse-amplitude-modulated) measuring system. The results showed that variations in temperature and salinityhad significant effects on tetraspore yield. However, variations in the photon flux density (from 15 μmol m⁻² s⁻¹ to 480 μmol m⁻² s⁻¹) had no apparent effect on tetraspore yield. Moreover, the PAM-parameters Y(I), Y(II), ETR(I), ETR(II) and F _v/F _m of colonies formed from different branches showed the same trend: parameter values of first generation branches>second generation branches>third generation branches. These results suggest that the photosynthetic activities of different colonies of branches changed with the same trend. Furthermore, photosynthesis in G. lemaneiformis was found to be involved in vegetative reproduction and tetraspore formation. Finally, the first generation branches grew slowly, but accumulated organic compounds to form large numbers of tetraspores. Taken together, these results showed that the first generation branches are ideal materials for the release of tetraspores.     

Expression of the Phycoerythrin Gene of Gracilaria lemaneiformis (Rhodophyta) in E. coli and Evaluation of the Bioactivity of Recombinant PE

Phycoerythrin (PE) is one of the most important proteins involved in light capturing during photosynthesis in red algae. Its potential biological activities had gained wide concerns. In the present study, tumor cytotoxic and hydroxyl radical assay were preformed to detect the bioactivity of recombinant PE. Recombinant plasmids pGEX-PE and pBGL were transformed into E.coli BL21 to make two recombinant strains BEX (pGEX-PE) and BGL (pBGL). PE expressing in BEX (pGEX-PE) was validated by SDS-PAGE and Western blotting analysis. SDS-PAGE analysis indicated that the PE-GST fusion protein was mostly inclusion bod- ies. Specific expression of PE was confirmed by Western blotting analysis. The recombinant E. coli BEX (pGEX-PE) cells were col- lected and sonicated. The supernatants were reserved for the tumor cytotoxic experiments. The result of tumor cytotoxic assay indi- cated that the supernatants containing PE had the activity of inhibiting the growth of Hela cells and with the increase of protein con- centration, the inhibiting rate increased from 37.31% to 63.26%, which showed significant difference from the control. Hydroxyl radical scavenging effect was tested with supernatants of BEX (pGEX-PE) and BGL (pBGL) cell lysates treated with sonication and heating. For the sonication samples, the scavenging rates of the supernatants of BEX (pGEX-PE) and BGL (pBGL) cell lysates were significantly higher than the negative control BL21(pGEX-4T) (P<0.02), and the scavenging rates increased slowly following the increase of the protein content. For the heating samples, except for the 0.2 mg mL-1 BGL (pBGL) products, the scavenging effects of the supernatants of BEX (pGEX-PE) and BGL (pBGL) cell lysates were stronger than that of negative control BL21(pGEX-4T). However, the effect intensity was not positively correlated with the increase of the protein concentration. Though a partially de- creased hydroxyl radical scavenging activity was led by heating, the biological activity was still retained and conspicuous. This re- search showed that phycoerythrin protein expressing in E. coli has the potential medical and sanitarian value.     

汞离子对细基江蓠孢子萌发和生长的影响

研究了4种Hg^2＋浓度对细基江蓠果孢子和四分孢子附着、萌发和生长的影响。结果表明：与对照组相比．Hg^2＋抑制了果孢子附着却促进了四分孢子的附着，但0．0025mg／L浓度与对照组的差异不显著（P〉0．05）．各浓度间差异显著（P〈O．05）；Hg^2＋在实验浓度范围内（O．0025、0．0050、0．0075、0．0100mg／L）影响了孢子的正常萌发，使孢子不能正常分裂和生长．刺激了孢子脱落，并且使盘状体的形态发生变异；Hg^2＋不恫浓度之间也存在显著差异（P〈0．05）．并且Hg^2＋浓度越高对孢子的萌发和生长影响越大。     

汞离子对细基江蓠孢子萌发和生长的影响

研究了4种Hg2+浓度对细基江蓠果孢子和四分孢子附着、萌发和生长的影响。结果表明:与对照组相比,Hg2+抑制了果孢子附着却促进了四分孢子的附着,但0.0025 mg/L浓度与对照组的差异不显著(P>0.05),各浓度间差异显著(P<0.05);Hg2+在实验浓度范围内(0.0025、0.0050、0.0075、0.0100 mg/L)影响了孢子的正常萌发,使孢子不能正常分裂和生长,刺激了孢子脱落,并且使盘状体的形态发生变异;`Hg2+不同浓度之间也存在显著差异(P<0.05),并且Hg2+浓度越高对孢子的萌发和生长影响越大。     

The Effect of Hydrolysis with Neutrase on Molecular Weight,Functional Properties,and Antioxidant Activities of Alaska Pollock Protein Isolate

In this study, the Alaska pollock protein isolate(APPI) was hydrolyzed by Neutrase for 20, 40, 80, 120, 160, 200, and 240 min. Hydrolysates with different molecular weights were produced and they were named as H1–H7. Furthermore, the effects of hydrolysis on the average molecular weights, functional properties(solubility, oil-holding capacities, foaming activities, and emulsifying properties), and antioxidant activities(1, 1-diphenyl-2-picrylhydrazyl, superoxide, and hydroxyl free radical-scavenging activities) were determined. It was found that when the degree of hydrolysis(DH) increased, the average molecular weights of the hydrolysates decreased significantly. The functional properties of APPI were also significantly improved. The hydrolysates of APPI exhibited better solubility, emulsifying activities, and foaming activities. Hydrolysates with low molecular weights(1 kDa) had better solubility, oil-holding capacities, and emulsifying activities, while hydrolysates with higher molecular weights(1 kDa) had better foaming activities. In addition, the hydrolysates exhibited excellent antioxidant properties, while the inhibition values of 1, 1-diphenyl-2-picryl hydroxyl(DPPH), superoxide, and hydroxyl free radical-scavenging activities, were 85.22%, 53.56%, and 75.00% respectively, when the concentration of the hydrolysates was 5.0 mg mL~(-1). The lower the average molecular weight was, the higher was the antioxidant activity. These results indicated that hydrolysis with Neutrase is an effective method for improving the functional and antioxidant properties of APPI. The hydrolysates of APPI displayed great potentials to be used as natural antioxidants in protein-rich aqueous foods such as nutrient supplements and sports beverages.     

远东拟沙丁鱼抗氧化肽对Caco-2细胞氧化应激损伤的影响

对远东拟沙丁鱼(Sardinops sagax)蛋白进行蛋白酶解和膜分离,得到分子质量100~3 000 u(F1)、3 000~10 000 u(F2)和大于10 000 u(F3)的多肽组分。在生化水平上测定各组分对1,1-二苯基-2-三硝基苯肼(DPPH)、2,2-联氮-二(3-乙基-苯并噻唑-6-磺酸)二铵盐(ABTS)、羟基自由基和过氧自由基清除能力;在细胞水平上,以H2O2对Caco-2细胞的毒性及细胞内抗氧化体系的影响为检测指标,筛选最适浓度的H2O2建立细胞氧化应激模型;在建立该模型的基础上,进一步检测蛋白多肽F1对氧化应激损伤细胞的修复功能。结果表明:组分F1对DPPH、ABTS、羟基自由基和过氧自由基清除能力最强,清除率分别为56.03±0.14、86.26±0.32、38.34±1.70、54.85±0.75;100μmol/L的H2O2对细胞毒性作用较小,并可降低细胞中谷胱甘肽过氧化物酶(GSH-Px)、总抗氧化物歧化酶(T-SOD)、过氧化物酶(POD)的活性和谷胱甘肽(GSH)的含量,增加丙二醛(MDA)的含量,细胞氧化应激模型选择H2O2浓度为100μmol/L;在该模型条件下,F1对受损的Caco-2细胞中抗氧化体系中的GSH-Px、T-SOD、POD的活性和MDA、GSH的含量有一定恢复作用,可调节细胞中失衡的抗氧化体系。远东拟沙丁鱼多肽FI对细胞氧化应激受损有一定保护作用。     

NaClO法漂白江蓠时不同工艺对氯气浓度及漂白效果的影响

为解决NaClO法漂白江蓠时漂白空间氯气浓度过高导致环境的污染。对NaClO法漂白江蓠时影响漂白空间氯气浓度的几种主要因素进行了研究。通过改进生产工艺 ,对酸化后的江蓠进行水洗后再漂白 ,同时增加草酸作后处理 ,在保证江蓠漂白效果的前提下 ,使漂白空间氯气浓度从最高值 80 5 4mg m3 降低到约 0 0 5 0mg m3 ,车间及环境的氯气浓度远低于国家安全标准 ,解决了江蓠漂白中氯气污染的问题 ,达到了清洁生产的效果。     

Antitumor and Antifungal Activities of Organic Extracts of SeacucumberHolothuria atra from the Southeast Coast of India

In phylum Echinodermata, the family Holothuridae is distinguished by its capacity of bioactive compounds. Sea cucumber Holothuria atra is commonly known as the lollyfish. The antifungal activity was detected using agar well diffusion method against the various fungal strains such as Trichoderma viride, Aspergillus niger, Aspergillus flavis, Candida albicans and Penicillium chrysogenum. Relatively high antifungal activity was seen against Candida albicans at 100 μL-1 concentration of extracts. Zone of inhibition was measured at 18 mm of diameter. The anti-tumor activities were detected against the Vero and Hep2 cell lines using MTT assay. The cells were treated with H. atra extract at concentrations 0.078-10 mg m L-1. The extract showed high proliferative activity against the Hep2 cells. The body wall extracts of sea cucumber(H. atra) showed effective antifungal and antitumor activities. All these findings suggest that the extracts could be used for the development of drugs.     

温度和盐度对军曹鱼幼鱼生长与抗氧化酶活性的影响

在实验室条件下,采用两因素交叉分组的方法,初步研究了环境因子(温度和盐度)对军曹鱼幼鱼肌肉抗氧化酶比活力的影响。温度设3个水平(26、29、32),盐度设4个水平(11、19、27、35),两个因素按其各自水平共产生12种组合。军曹鱼幼鱼在玻璃钢水槽内养殖20d后取样测定超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和谷胱甘肽过氧化物酶(GPX)的比活力。结果表明:盐度因子诱导了军曹鱼的抗氧化应激反应,抗氧化酶活力随盐度降低而升高,SOD、CAT和GPX比活力分别从35时的17.74±3.38、19.44±2.66和13.70±2.99U/mg上升到11时的30.14±1.76、35.63±10.59和17.44±4.32U/mg;温度、盐度及因素间的交互作用对SOD和CAT比活力有显著影响(P<0.05);温度对GPX比活力无显著影响(P>0.05),而盐度及因素间的交互作用对GPX比活力影响显著(P<0.05)。     

Hg-Cd复合胁迫对卵叶喜盐草光合色素和丙二醛含量及抗氧化酶活性的影响

通过胁迫试验研究Hg-Cd复合胁迫对卵叶喜盐草光合色素含量、丙二醛(MDA)含量及抗氧化酶活性的影响。结果表明,Hg-Cd复合毒性对卵叶喜盐草叶绿素a、叶绿素b和类胡萝含量显著影响(P0.05),当Hg、Cd质量浓度分别大于0.005和0.100 mg·L~(-1)时,叶绿素a、b比值,叶绿素a和类胡萝卜素含量下降,光合作用受抑制;Hg-Cd复合胁迫对卵叶喜盐草的MDA含量影响有统计学意义(P0.05),当Hg、Cd质量浓度分别为0.005和0.100 mg·L~(-1)时,MDA含量上升41.5%;Hg、Cd复合胁迫对卵叶喜盐草的超氧化物歧化酶(SOD)、过氧化物酶(POD)和过氧化氢酶(CAT)活性影响有统计学意义(P0.05),SOD活性随浓度梯度逐渐下降,在Hg、Cd质量浓度分别大于0.010和0.200 mg·L~(-1)时,SOD活性降低,POD和CAT活性随胁迫浓度梯度呈先增后减趋势,其中当Hg、Cd质量浓度分别为0.010和0.20 mg·L~(-1)时,而POD和CAT活性均上升为最大值,分别较对照组增加12.35%和22.75%,当Hg、Cd质量浓度分别大于0.015和0.300 mg·L~(-1)时,POD和CAT活性开始下降至最低。总的来说,Hg、Cd联合胁迫的浓度大于0.005和0.100 mg·L~(-1)时,卵叶喜盐草光合色素含量下降,MDA含量和抗氧化酶活性均相应发生变化。     

雌激素的抗肿瘤效果和小鼠腹腔巨噬细胞的雌激素受体水平(英文)

探索肿瘤生长性别差异的原因。研究雌激素及其拮抗剂他莫昔芬对可移植性小鼠肉瘤180生长的影响;采用同位素方法,证实了完整小鼠腹腔巨噬细胞对3H-雌二醇的特异性细胞内结合。肯定了小鼠腹腔巨噬细胞是一种雌激素受体阳性细胞。雌性小鼠肿瘤生长速率比雄性小鼠低是因为雌性激素激活了雌激素受体阳性的巨噬细胞的缘故。     

亚硝酸氮对红鳍东方鲀的毒性效应

采用半静水生物测试法,研究亚硝酸盐氮对红鳍东方鲀(Takifugu rubripes)(9.5⁹.8 g)的急性毒性及对部分肝脏抗氧化酶活性的影响。结果表明,在急性毒性实验中,亚硝酸盐氮对红鳍东方鲀的24、48、72和96 h半致死质量浓度(LC50)分别为201.88、190.57、171.09、159.25 mg/L,安全质量浓度为15.92 mg/L;在亚急性毒性实验中,当红鳍东方鲀分别在5.3、10.6,15.9、21.2 mg/L亚硝酸盐氮中暴露4周时,其肝脏总抗氧化能力(T-AOC)、超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性随亚硝酸盐氮浓度的升高均被显著抑制,差异有统计学意义(P<0.05),表明亚硝酸盐氮会对鱼体的抗氧化能力产生影响。     

亚硝酸氮对红鳍东方[鱼屯]的毒性效应

采用半静水生物测试法,研究亚硝酸盐氮对红鳍东方[鱼屯]（Takifugu rubripes）（95～9．8g）的急性毒性及对部分肝脏抗氧化酶活性的影响。结果表明,在急性毒性实验中,亚硝酸盐氮对红鳍东方[鱼屯]的24、48、72和96h半致死质量浓度（LC50）分别为20188、19057、17109、15925mg／L,安全质量浓度为1592mg／L;在亚急性毒性实验中,当红鳍东方纯分别在5．3、106,159、212mg／L亚硝酸盐氮中暴露4周时,其肝脏总抗氧化能力（T—AOC）、超氧化物歧化酶（SOD）和过氧化氢酶（CAT）活性随亚硝酸盐氮浓度的升高均被显著抑制,差异有统计学意义（P〈0.05）,表明亚硝酸盐氮会对鱼体的抗氧化能力产生影响。     

Effect of Zinc Oxide Nanoparticles on Denitrification and Denitrifying Bacteria Communities in Typical Estuarine Sediments

For revealing the effects of increasing of zinc oxide nanoparticles(ZnO NPs) on denitrification and denitrifying bacteria communities in estuarine sediments, the surface sediments of two typical estuaries(the Yangtze River Estuary and the Yellow River Estuary) were added with medium concentration(170 mg L~(-1)) and high concentration(1700 mg L~(-1)) of ZnO NPs for anaerobic culture in laboratory. The concentration of NO_3~-and NO_2~-, the reductase activity and denitrification rate were measured by physicochemical analysis, nir S gene abundance and denitrifying bacteria communities by molecular biological methods. The results showed that ZnO NPs inhibited NO_3~-, NO_2~-reduction process and NO_3~-, NO_2~-reductase activity, and a stronger inhibition effect resulting from the higher ZnO NPs concentration. ZnO NPs decreased nirS gene abundance and community diversity of denitrifying bacteria. In addition, the inhibition degree of ZnO NPs on the denitrification process of sediments in different estuaries was different. These results were of great significance for evaluating the potential ecological toxicity and risks of nanomaterials in estuaries.     

Effects of 5-Azacytidine(AZA) on the Growth,Antioxidant Activities and Germination of Pellicle Cysts of Scrippsiella acuminata(Dinophyceae)

In this study,we investigated the effects of different concentrations of 5-azacytidine (AZA),a DNA methyltransferase inhibitor,on the growth,antioxidant activities and germination of pellicle cysts of Scrippsiella acuminata.The purpose of this study is to understand the toxic effects of AZA on marine microalgae,and to demonstrate the effect of DNA methyltransferase inhibitors on the germination of pellicle cysts.Results showed that AZA inhibited the growth of S.acuminata significantly,and displa...     

Effects of Silver Nanoparticles on Denitrification and Associated N2O Release in Estuarine and Marine Sediments

Silver nanoparticles(AgNPs)have been widely used in medicine and consumer products.And it enters the river in dif-ferent ways,then finally converges to the ocean through the estuary.AgNPs polution can affect NO₂?and N₂O production by denitri-fiers in aquatic system.The effects of AgNPs on denitrification activity,nitrogen transformation and nitrous oxide(N₂O)emission were investigated in Dagu River Estuary(DRE)and Jiaozhou Bay(JZB).The results showed that the potential denitrification activity(PDA),NO₃?and NO₂?reduction rates decreased with an increase of AgNPs concentration in DRE and JZB.However,the N₂O ac-cumulation was significantly increased at AgNPs concentrations above 5 mg kg^?1in both areas,and the accumulation rate was greater in estuary than in bay(P<0.05).Moreover,the total bacterial count showed a slightly increasing trend with an increase of AgNPs concentration(P>0.05)in DRE and JZB.Importantly,the relative abundance of narG,nirS and nosZ gene in two areas decreased with the increase of AgNPs concentration,and the negative effect of AgNPs varied in order:nosZ>nirS>narG,inferring that the expression of denitrifying related genes could be significantly and differently inhibited by AgNPs addition.Thus,this study demon-strated that the inhibitory effect of AgNPs on different denitrification process,which may lead to the increase of inorganic nitrogen accumulation and N₂O realease.This study provides a scientific basis for the further studies of AgNPs on the ecological impact mechanism and environmental effects of offshore sedimentary environment.     

In vitro Antioxidant Effects of Porphyra haitanensis Peptides on H_2O_2-Induced Damage in HepG2 Cells

In this study,protein from Porphyra haitanensis was used as raw material to prepare an antioxidant peptide,and its antioxidant activity was evaluated in vitro.A model of H_2O_2-induced oxidative damage in Hep G2 cells was established,and the effects of Porphyra haitanensis hydrolysates (PHHs) on superoxide dismutase (SOD) activity and malondialdehyde (MDA) content were detected.Finally,the structure of PHHs was identified by ESI-MS/MS.The results showed that the 1,1-diphenyl-2-pyridylhydrazine(DPPH)-free radical-scavenging ability of PHHs was the strongest (59.28%at 1.0 mg m L~(-1)) when hydrolyzed with an acidic protease for 4 h.PHHs with different concentrations had protective effects on H_2O_2-induced damage to Hep G2 cells,and the protective effect was enhanced with increasing concentrations.When the level was 400μg m L~(-1),the cell survival rate was as high as 88.62%.Moreover,PHHs can significantly reduce oxidative damage to Hep G2 cells by H_2O_2,improve SOD activity,and reduce MDA content.The tetrapeptide Asp-Lys-Ser-Thr,with a molecular weight of 448 Da,was identified as an important fraction of PHHs by high-resolution mass spectrometry.     

Effects of temperature on photosynthetic performance and nitrate reductase activity in vivo assay in Gracilariopsis lemaneiformis (Rhodophyta)

Journal of Oceanology and Limnology - Gracilariopsis lemaneiformis is an economically-valued species and widely cultured in China at present. After being acclimated to different growth temperatures...