N-TiO2/硅藻土负载型纳米材料可见光催化降解水中Microcystin-LR

以钛酸四丁酯(C16H36O4Ti)为Ti源,尿素((NH2)2CO)为N源,硅藻土(Diatomite)为载体,采用溶胶凝胶法制备硅藻土负载氮掺杂二氧化钛改性纳米材料(N-TiO2/Diatomite).利用SEM、XRD、XPS、FT-IR、UV-Vis DRS对其进行系列特性表征及可见光催化性能分析,探究Microcystin-LR初始浓度对光催化的影响,分析其降解动力学和降解途径.结果表明,硅藻土负载优化了N-TiO2分子的分散性,形成了链条型纳米孔隙球状结构;N元素掺杂、硅藻土负载不影响材料晶型,晶粒粒径减小至11.39 nm;Si、N分别与TiO2成键;光响应强度和范围红移,可见光活性显著提升.以优化制备条件(350℃煅烧、掺N的物质的量分数为8%)的改性N-TiO2/Diatomite材料可见光催化降解Microcystin-LR,当初始质量浓度为1 mg·L^-1的Microcystin-LR水溶液反应6 h后降解率可达95.0%,矿化率为83.9%;反应符合准一级动力学方程,速率常数k为0.453 3 h^-1.

微囊藻毒素Microcystin-LR体外遗传毒性

背景与目的:应用人类淋巴母细胞TK6研究微囊藻毒素(Microcystin_LR,MCLR)的体外遗传毒性。材料与方法:MCLR体外染毒TK6细胞4h或24h后检测细胞毒性、微核及tk位点突变频率。结果:4h染毒未引发明显细胞毒性,24hMCLR染毒导致TK6细胞相对存活率下降,细胞微核率及TK基因突变频率明显上升,并有剂量-反应关系。最高浓度组(80μg/ml)的细胞微核率及TK基因突变频率分别是对照组的4.8及5.1倍。MCLR诱发tk位点两种不同表型的突变细胞集落,即正常生长集落及缓慢生长集落,并以后者为主。结论:24h染毒MCLR可以诱发TK6细胞微核及基因突变,揭示MCLR可能是一种断裂剂。

Using the nematode Caenorhabditis elegans as a model animal for assessing the toxicity induced by microcystin-LR

Among more than 75 variants of microcystin （MC）, microcystin-LR （MC-LR） is one of the most common toxins. In this study, the feasibility of using Caenorhabditis elegans to evaluate MC-LR toxicity was studied. C. elegans was treated with MC-LR at different concentrations ranging from 0.1 to 80 μg/L. The results showed that MC-LR could reduce lifespan, delay development, lengthen generation time, decrease brood size, suppress locomotion behavior, and decreases hsp-16-2-gfp expression. The endpoints of generation time, brood size, and percentage of the population expressing hsp-16-2-gfp were very sensitive to 1.0μg/L of MC-LR, and would be more useful for the evaluation of MC-LR toxicity. Furthermore, the tissue-specific hsp-16-2-gfp expressions were investigated in MC-LR-exposed animals, and the nervous system and intestine were primarily affected by MC-LR. Therefore, the generation time, brood size, and hsp-16-2-gfp expression in C. elegans can be explored to serve as valuable endpoints for evaluating the potential toxicity from MC-LR exposure.

Microcystin-LR对体外牛子宫内膜上皮细胞增殖和凋亡的影响

使用Microcystin-LR体外处理牛子宫内膜上皮细胞,探究其对细胞活率影响及分子调控机制,以期为提高牛繁殖效率提供理论基础和实践依据。试验运用CCK-8检测MC-LR处理后细胞存活率;流式细胞术检测细胞周期变化和细胞凋亡率;Annexin V-FITC试剂盒检测细胞凋亡;实时荧光定量检测Pi3k、Akt、mTOR、Cyt-c、Caspase-3、Caspase-9和p53基因mRNA表达量。Western blot检测Pi3k、Akt、Caspase-3和Caspase-9蛋白表达量。结果表明,不同浓度和时间处理下细胞存活率出现升高和降低两种相反情况,根据CCK-8检测结果,选择12 h,100μg·L^(-1)和24 h,160μg·L^(-1)两种处理方案开展后续试验。在12 h,100μg·L^(-1)条件下,Pi3k、Akt、mTOR的mRNA表达量升高(P<0.01),Cyt-c、Caspase-3和Caspase-9基因mRNA表达量下降(P<0.05),Pi3k和Akt蛋白表达量升高(P<0.05),Caspase-3和Caspase-9蛋白表达量降低(P<0.05)。处于G0/G1期细胞数量减少(P<0.01),处于G2/M和S期细胞数量增加(P<0.01);p53基因mRNA表达量升高(P<0.05)。在24 h,160μg·L^(-1)处理条件下,Caspase-3、Caspase-9、Bax/Bcl2基因mRNA表达量升高(P<0.05),Caspase-3和Caspase-9蛋白表达量升高(P<0.05)。研究表明,100μg·L^(-1)MC-LR处理细胞12 h,细胞通过增强Pi3k-Akt-mTOR信号通路,抑制线粒体凋亡通路促进细胞增殖,同时导致处于G1期细胞数量减少、G2/M期和S期细胞数量增加。160μg·L^(-1)MC-LR处理细胞24 h,细胞通过活化线粒体凋亡通路发生凋亡。

Studies on Extracting Microcystin-Lr from Microcystis Aeruginosa by Water Bath

Different temperatures of water bath was used to extract the intracellular microcystin-LR(MC-LR) of Microcystis aeruginosa. Researching the extraction efficiency under the suitable temperature, so that it could find out the best temperature and time for extracting MC-LR from Microcystis aeruginosa cells. Five equal Microcystis aeruginosa was used to find out the best temperature, extracting at 60℃, 70℃, 80℃, 90℃ and 100℃ for 15 minutes, respectively. Results showed that the content of MC-LR extracted with the water under 100℃ was the highest. But meanwhile, the type and the content of impurities was the highest, too. In addition, another six equal Microcystis aeruginosa was extract with the water under 100℃ for 5min, 10 min, 15 min, 20 min, 25 min and 30 min respectively. It was proved that 20 minutes was enough for extracting MC-LR from Microcystis aeruginosa, no long time was needed.

Photocatalytic Treatment of Microcystin-LR-Containing Wastewater Using Pt/WO ₃Nanoparticles under Simulated Solar Light

This study investigates the photocatalytic degradation of microcystin-LR (MC-LR) under simulated solar light using Pt modified nano-sized tungsten trioxides (Pt/WO3). Photocatalytic activity was higher during the degradation of MC-LR with Pt/WO 3 than with pure WO 3 or Ti O2 . The catalyst loading greatly affect the degradation performance. The rate of degradation is influenced by the initial pH of the reaction solution. This study also investigates the photocatalytic inactivation of cyanobacteria. The results show that the algal growth was successfully controlled by the Pt/W O 3 . This study suggests Pt/W O 3 photocatalytic oxidation with solar light is a promising treatment for water containing MC-LR.

Degradation of Microcystin-LR by Gas-Liquid Interfacial Discharge Plasma

In this study, we report on the degradation of microcystin-LR （MC-LR） by gas- liquid interracial discharge plasma. The influences of operation parameters such as average input voltage, electrode distance and gas flow rate are investigated. Experimental results indicate that the input voltage and gas flow rate have positive influences on MC-LR degradation, while the electrode distance has a negative one. After 6 min discharge with 25 kV average input voltage and 60 L/h air aerati by discharge both in on, the degradation rate of MC-LR achieves 75.3%. distilled water and MC-LR solution are measured H202 and 03 generated Moreover, an emission spectroscopy is used as an indicator of the processes that take place on the gas-liquid boundary and inside plasma. Varied types of radicals （O, .OH, CO, 03, etc.） are proved to be present in the gas phase during gas-liquid interfacial discharge.

The expression of Bcl-2 and Bax produced by sub-chronic intoxication with the cyanotoxin Microcystin-LR

Objective:We explored the role and molecular mechanism of Microcystin-LR(MC-LR) in hepatocarcinogenesis.Methods:The two-stage-medium-term theory was applied to the establishment of the animal model.The promoting effect of MC-LR on liver tumor was evaluated with the Albert γ-GT methods.During the tumor-promoting course,the effects of MC-LR on the regulation and expression of the Bcl-2 and Bax genes were studied with immunohistochemical technique and RT-PCR.Results:MCLR could enhance the positive reaction of Albert γ-GT(GGT),a preneoplasm marker.The positive reaction rate of GGT in diethylnitrosamine(DEN) + MC group was significantly higher than that in the DEN control group.The protein and RNA expression of Bcl-2 was significantly increased by MC-LR,and that the protein expression of Bax was decreased simultaneously.Conclusion:These results indicate that the MC-LR can inhabit apoptosis through the Bcl-2 and Bax genes.Therefore,we conclude that the expression change of Bcl-2 and Bax genes possibly plays an important role in the promotion of liver tumor by MC-LR.

Removal of Microcystin-LR in Water by Chlorine Dioxide

Microcystins （ MCs ） are well known as hepatotoxins produced by blooms of toxic cyanobacteria （blue-green algae） abundant in surface water used as drinking water resource and have drawn attention of environmentalists world over by leading to adverse health effects. A study on efficiency and reaction kinetics of microcystin-LR （ MC-LR ） degradation by CIO2 was performed. Experimental results indicated that MC-LR was removed by CIO2 effectively and the residual concentration of MC-LR could meet the national guideline（GB5749 - 2006） （1.0 μg · L^-1）, the efficiency of removal was in positive correlation to CIO2 dosage and reaction time and in negative correlation to initial concentration of MC-LR and pH value, whereas it was affected by temperature slightly. CIO2 dosage was the most important reaction factor on base of the orthogonal test results. The reaction was second order overall and first order with respect to both CIO2 and MC- LR, and had an activation energy of 78.81 kJ · mo1^-1 . The reaction rate constant was 4.74× 10-^2 L/（mol · min） at 10 ℃. Therefore, oxidation of CIO~ could be taken as an effective technology for removing MC-LR from drinking water resources in traditional drinking water supplies.

Photochemical Reactions of Microcystin-LR Following Irradiation with UV Light

Photochemical reactions of microcystin-LR, a toxic compound produced by some blue green algae, were investigated. Ultraviolet absorption of microcystin-LR was assessed. Time-dependent density functional theory (TDDFT) calculations indicated that absorption peak at 238 nm was mainly due to excitation of electrons from the linear chain structure Adda of microcystin-LR. Irradiation of microcystin-LR with UV light resulted in the reduction of the 238 nm absorption peak and the appearance of a new peak at 300 nm. Density functional theory (DFT) and TDDFT calculations with a model molecule suggested that this 300 nm peak was due to tricyclo-Adda microcystin-LR, an intermediate in photochemical reactions of microcystin-LR. Analysis of the rate of this photochemical reaction showed that it was a first order reaction.

Removal of Microcystin-LR in lake water sample by hydrophilic mesoporous silica composites under high-throughput MALDI-TOF MS detection platform

Microcystins(MCs),a family of cyclic heptapeptide cyanotoxins,exists in aquatic environment where cyanobacterial bloom happens,which will accumulate in aquatic organisms and transfer through the food chain to higher trophic levels,posing a health risk to both animals and human bodies.Among various MCs,Microcystin-LR(MC-LR)is worthiest studied for its strong toxicity,ubiquity and widespread.Here in this work,iminodiacetic acid(IDA)decorated magnetic mesoporous silica(mSiO_(2))nanocomposites(Fe_(3)O_(4)@mSiO_(2)-IDA)were facilely synthesized which possessed the merits of large surface area(188.21 m^(2)/g),accessible porosity(2.66 nm),excellent hydrophilicity and rapid responsiveness to magnetic field.Then the composites were successfully employed to the removal process of Microcystin-LR in real water samples followed by Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry(MALDI-TOF MS)analysis,achieving the removal efficiency above 92.5%even after ten recycles of the composites.It provided a potential method for removing MC-LR in aqueous environment with high effectiveness,lower costs and less secondary contamination.

Serum Microcystin-LR Levels Linked with Risk of Inflammatory Bowel Disease:A Matched Case-Control Study in China

Microcystin-LR(MC-LR),the most prevalent and diverse cyanotoxin produced by harmful cyanobacterial blooms,has been linked to gastrointestinal toxicity.Therefore,we conducted a case-control study across four regions in China to investigate this relationship.Inflammatory bowel disease(IBD)cases(219)were matched with healthy controls(438)based on age and gender and conditional logistic regression models and Restricted cubic splines were used to evaluate the association between MC-LR exposure and IBD risk.We used quantitative real-time polymerase chain reaction to measure the expression levels of inflammatory factors.The levels of protein expression in the colorectum were determined using Western blotting(WB).Compared to the lowest quartile of serum MC-LR levels,the adjusted odds ratios and 95%confidence intervals(CI)for the highest quartiles of serum MC-LR levels were 5.51(2.70,11.21).The RCS was shown the association between serum MC-LR levels and IBD risk was nonlinear(Pnonlinear<0.001).In the animal experiments,MC-LR resulted in colorectal injury via the PI3K/AKT signaling pathway.Our study provides the evidence that serum MC-LR exposure is significantly associated with the risk of IBD in China.Animal study results indicate that MC-LR probably causes IBD via the PI3K/AKT signaling pathway.

氨氮和微囊藻毒素-LR联合作用对斑马鱼肠道免疫和菌群的影响

氨氮和微囊藻毒素-LR(MCLR)是水生环境中普遍存在的污染物。为探讨两者对斑马鱼肠道潜在的协同效应,实验将成年雌性斑马鱼分别暴露于氨氮(30 mg·L^(-1))、MCLR(10μg·L^(-1))以及两者混合(30 mg·L^(-1)+10μg·L^(-1))的环境中,持续30 d。组织病理学分析显示:氨氮暴露导致肠绒毛面积减少;MCLR暴露导致肠道绒毛破裂,空泡化面积增加;而联合暴露对肠组织损伤更为严重。这些变化伴随着肠道中溶菌酶和β-防御素的含量及相关基因表达的显著降低,表明斑马鱼肠道免疫功能受到抑制。此外,氨氮和MCLR的单独及联合处理还激活NOD1/2和TLR4a/4b信号通路,导致促炎因子IL-1β和TNF-α的表达水平和蛋白含量上升,进而可能诱发肠道炎症反应。肠道菌群分析结果进一步显示,氨氮和MCLR处理显著改变斑马鱼肠道内菌群的平衡,即氨氮增加厚壁菌门(Firmicutes)丰富度,MCLR增加放线菌门(Actinobacteria)丰富度但降低变形菌门(Proteobacteria)丰富度,而氨氮和MCLR联合作用增加肠道致病菌群假单胞菌属(Pseudomonas)和分枝杆菌属(Mycobacterium)丰富度。进一步,两者联合暴露还导致肠道中产生短链脂肪酸的菌群丰度和短链脂肪酸含量显著降低。综上所述,氨氮和MCLR联合处理对斑马鱼肠道免疫及菌群稳态产生了协同的负面影响,其对水生动物和水生态系统的健康构成了不容忽视的潜在风险。

α-硫辛酸在微囊藻毒素-LR诱导草鱼卵巢细胞损伤中的作用

为研究α-硫辛酸(Alpha lipoic acid,α-LA)在微囊藻毒素-LR(Microcystin-LR,MC-LR)诱导水生动物毒性效应中的保护作用,本研究以草鱼卵巢(Grass carp ovary,GCO)细胞为研究对象,检测分析了不同浓度α-LA以及α-LA与MC-LR共同暴露对GCO细胞活力的影响,随后根据测定的结果设置对照组(不添加MC-LR和α-LA)、125μmol·L^(-1)α-LA组、24μmol·L^(-1)MC-LR组及125μmol·L^(-1)α-LA+24μmol·L^(-1)MC-LR组,检测分析了α-LA在缓解MC-LR对GCO细胞活性、氧化应激以及炎症方面的影响。结果表明:与对照组相比,24μmol·L^(-1)MC-LR可诱导乳酸脱氢酶(LDH)活性和丙二醛(MDA)含量显著上升,同时显著抑制谷胱甘肽(GSH)活性(P<0.05),当MC-LR处理组中加入125μmol·L^(-1)α-LA后,与MC-LR单独暴露组相比,LDH活性和MDA含量均显著下降(P<0.05),但GSH活性却显著上升(P<0.05)。对氧化相关基因分析发现,与对照组相比,24μmol·L^(-1)MC-LR可显著降低SOD1、CAT和GST基因的表达量(P<0.05),当125μmol·L^(-1)α-LA与24μmol·L^(-1)MC-LR共同作用于GCO细胞后,与MC-LR单独暴露组相比,联合暴露组的GST基因的表达量显著上升(P<0.05),但SOD1和CAT两个基因的表达变化不显著(P>0.05)。此外,对炎症因子进行分析发现,MC-LR单独暴露组TNFα和IL11基因的相对表达水平显著高于对照组(P<0.05),但联合暴露组中的TNFα和IL11基因的相对表达水平却显著低于MC-LR单独暴露组(P<0.05)。研究结果表明,α-LA可缓解MC-LR诱导的氧化应激,提高细胞活力,抑制GCO细胞炎症的发生,从而减弱MC-LR对GCO细胞的损伤。

Visible light-induced N-doped TiO_2 nanoparticles for the degradation of microcystin-LR

N-doped nano-crystalline TiO2 powders have been synthesized by the sol-gel method.The shape and crystal structure of the resulting N-doped TiO2 were investigated by X-ray Photoelectron Spectroscopy (XPS),X-ray spectroscopy (XRD),Transmission Electron Microscopy (TEM) and UV-vis reflection spectrum.The results showed that doping TiO2 with nitrogen can lower its band gap and apparently shift its optical response to the visible region.Under the visible light (λ】 420 nm) irradiation,the MC-LR was degraded by the synthesized N-TiO2 nano-material.The variation of MC-LR amount and its intermediates were detected by high performance liquid chromatography (HPLC) and LC-MS,respectively.The mineralization of MC-LR was determined by total organic carbon (TOC) analysis.Simultaneously,transient oxidative species generated during photocatalysis were tracked by electron spin resonance (ESR) and Peroxidase method.All these results indicated that visible-light excited N-TiO2 can activate molecular oxygen and thereby achieve degradation of MC-LR completely within 14 h.The removal of 59% of TOC was achieved after 20 h irradiation.The major oxidative species in the system were hydroxyl radical (·OH) and H2O2.13 Kinds of intermediates were primarily identified in the process.Based on these results,a reasonable conclusion was drawn for the degradation of MC-LR wherein its four positions are easy to be attacked by the photo-generated OH radical followed by the hydrolyzation of peptides.

Removal of microcystin-LR from drinking water using a bamboo-based charcoal adsorbent modified with chitosan

A new kind of low-cost syntactic adsorbent from bamboo charcoal and chitosan was developed for the removal of microcystin-LR from drinking water. Removal effciency was higher for the syntactic adsorbent when the amount of bamboo charcoal was increased. The optimum dose ratio of bamboo charcoal to chitosan was 6：4, and the optimum amount was 15 mg/L; equilibrium time was 6 hr. The adsorption isotherm was non-linear and could be simulated by the Freundlich model （R2 = 0.9337）. Adsorption efficiency was strongly afflected by pH and natural organic matter （NOM）. Removal efficiency was 16% higher at pH 3 than at pH 9. Efficiency rate was reduced by 15% with 25 mg/L NOM （UV254 = 0.089 cm-1） in drinking water. This study demonstrated that the bamboo charcoal modified with chitosan can effectively remove microcystin-LR from drinking water.

Effect of microcystin-LR on protein phosphatase 2A and its function in human amniotic epithelial cells

Due to their toxicity,the increased distribution of microcystins(MCs) has become an important worldwide problem.MCs have been recognized as inhibitors of protein phosphatase 2A(PP2A) through their binding to the PP2A catalytic subunit.However,the exact mechanism of MC toxicity has not been elucidated,especially concerning the cellular response and its autoregulation.To further dissect the role of PP2A in MC-induced toxicity,the present study was undertaken to determine the response of PP2A in human amniotic epithelial(FL) cells treated with microcystin-LR(MCLR),one of the MC congeners.The results show that a low-dose treatment of MCLR in FL cells for 6 h induced an increase in PP2A activity,and a high-dose treatment of MCLR for 24 h decreased the activity of PP2A,as expected.The increased mRNA and protein levels of the PP2A C subunit may explain the increased activity of PP2A.Furthermore,MCLR altered microtubule post-translational modifications through PP2A.These results further clarify the underlying mechanism how MCLR affects PP2A and may be helpful for elucidating the complex toxicity of MCLR.

Point-of-care detection of Microcystin-LR with a personal glucose meter in drinking water source

In this study,a point-of-care sensing protocol has been reported for rapid and sensitive detection of Microcystin-LR(MC-LR)in water by personal glucose meter.The proposed immunosensor has been fabricated by using a primary antibody coated ZnFe2O4 nanoparticles to capture the target MC-LR.Consequently,the invertase@secondary antibody-conjugated graphene oxide-Au NPs can be immobilized for formating the sandwich immuno-complexes,which allowed for enzymatic conversion of sucrose to glucose.Thus,the concentration of MC-LR can be refelected by the converted glucose,which can be easily measured by the personal glucose meter(PGM).The PGM readout immunosensing method possessed good reproducibility and stability,which may have significant potential for other applications.

A competitive microcystin-LR immunosensor based on Au NPs@metal-organic framework(MIL-101)

An electrochemical immunosensor was developed for ultrasensitive detection of microcystin-LR in water. MIL-101, a porous metal-organic frameworks(MOFs) material based on trivalent chromium skeleton were synthesized by hydrothermal synthesis method, and loaded with Au nanoparticles(Au NPs) to prepare Au NPs@MIL-101 composite materials which were used as a marker to label anti microcystin-LR(Anti-MC-LR). The composite materials have strong catalytic properties to the oxidation of ascorbic acid. Anti-MC-LR was immobilized on glassy carbon electrode surface using electrodeposition graphene oxide(GO) as an immobilization matrix to construct a competitive microcystin-LR immunosensor. The electrochemical immunosensor display linear relationship in the range of 0.05 ng/mL-75 μg/mL with linear correlation coefficient of 0.9951 and detection limit of 0.02 ng/mL(S/N = 3). This sensor was used to detect microcystin-LR in the water sample. The recovery was 102.43%,which is satisfied. The good testing results indicate the sensor has a great prospect in practical application.

Experimental and model comparisons of H_2O_2 assisted UV photodegradation of Microcystin-LR in simulated drinking water

The degradation of Microcystin-LR (MC-LR) in water by hydrogen peroxide assisted ultraviolet (UV/H2O2) process was investigated in this paper. The UV/H2O2 process appeared to be effective in removal of the MC-LR. MC-LR decomposition was primarily ascribed to production of strong and nonselective oxidant-hydroxyl radicals within the system. The intensity of UV radiation, initial concentration of MC-LR, MC-LR purity, dosages of H2O2, the initial solution pH, and anions present in water, to some extent, influenced the degradation rate of MC-LR. A modified pseudo-first-order kinetic model was developed to predict the removal efficiency under different experimental conditions.