Hydroxyl radicals (*OH) are associated with titanium dioxide (TiO(2)) nanoparticle-induced cytotoxicity and oxidative DNA damage in fish cells

TiO(2) nanoparticles (< 100 nm diameter) have been reported to cause oxidative stress related effects, including inflammation, cytotoxicity and genomic instability, either alone or in the presence of UVA irradiation in mammalian studies. Despite the fact that the aquatic environment is often the ultimate recipient of all contaminants there is a paucity of data pertaining to the potential detrimental effects of nanoparticles on aquatic organisms. Therefore, these investigations aimed to evaluate the potential cytotoxic and genotoxic effects of TiO(2) nanoparticles on goldfish skin cells (GFSk-S1), either alone or in combination with UVA. Whilst neutral red retention (NRR) assay (a measure of lysosomal membrane integrity) was used to evaluate cell viability, a modified Comet assay using bacterial lesion-specific repair endonucleases (Endo-III, Fpg) was employed to specifically target oxidative DNA damage. Additionally, electron spin resonance (ESR) studies with different spin traps were carried out for qualitative analysis of free radical generation. For cell viability, TiO(2) alone (0.1-1000 microg ml(-1)) had little effect whereas co-exposure with UVA (0.5-2.0 kJm(-2)) caused a significant dose-dependent decrease which was dependent on both the concentration of TiO(2) and the dose of UVA administered. For the Comet assay, doses of 1, 10 and 100 microg ml(-1) in the absence of UVA caused elevated levels of Fpg-sensitive sites, indicating the oxidation of purine DNA bases (i.e. guanine) by TiO(2). UVA irradiation of TiO(2)-treated cells caused further increases in DNA damage. ESR studies revealed that the observed toxic effects of nanoparticulate TiO(2) were most likely due to hydroxyl radical (OH) formation.     

Enhancement of solar inactivation of Escherichia coli by titanium dioxide photocatalytic oxidation

AIMS: To improve solar water disinfection using a photocatalysing semi-conductor and to study the mechanisms involved in this process. METHODS AND RESULTS: Cells of Escherichia coli were used as the microbiological indicator to study the possibility of improving the efficiency of solar water disinfection using titanium dioxide (TiO2) as a photooxidizing semi-conductor. TiO2 was used either as a suspended powder or in an immobilized form. Both applications improved the efficiency of solar disinfection. TiO2 in suspension was more effective than the immobilized form, producing enhancement factors of 1.62 and 1.34, respectively. The concentration of TiO2 greatly affected efficiency, with a maximum effect at 1 mg ml(-1). Higher TiO2 concentrations reduced the efficiency. Dimethyl sulphoxide (DMSO) and cysteamine (Cys), hydroxyl radical (OH.) scavengers, were used to elucidate the mechanisms involved in the presence of TiO2. Both DMSO and Cys totally abolished the enhancing effect produced by the presence of TiO2. CONCLUSIONS: Sunlight has a potential water disinfecting capacity. The use of TiO2 greatly improved this efficiency. The effect of TiO2 was mainly concentration-dependent, giving maximum efficiency at 1 mg ml(-1). The presence of DMSO and Cys removed the TiO2-induced enhancement, indicating that OH. may be involved in the process of cell killing. SIGNIFICANCE AND IMPACT OF THE STUDY: The efficiency of solar disinfection is limited and time-consuming and needs to be improved. The use of a semi-conductor is promising as it reduces the time of exposure and therefore increases the efficiency of solar disinfection. This would allow for the availability of good quality water, and hence would improve the quality of life.     

沙棘果渣总黄酮提取工艺优化及抗氧化活性研究

利用果胶酶协同超声波法,对沙棘果渣有效成分总黄酮的提取工艺及其抗氧化活性进行了研究.以提取率为指标,通过酶用量、液料比、乙醇浓度、提取时间、提取温度、超声功率等单因素分析,选定酶用量、液料比、超声提取时间3个因素进行响应面试验,确定提取优化工艺条件为:果胶酶用量5.1％,液料比41∶1,超声提取时间81 min,此条件...     

Photocatalytic disinfection of Giardia intestinalis and Acanthamoeba castellani cysts in water

In this study, disinfection of water containing Giardia intestinalis and Acanthamoeba castellani cysts with TiO2 and modified catalyst silver loaded TiO2 (Ag-TiO2) was investigated. Destruction of the parasites was evaluated after UV illumination of the suspension consisting 5 x 10(8)-13.5 x 10(8)cysts/mL in the presence of 2g/L neat or modified TiO2 at neutral pH. In the initial stage, the solid photocatalyst particles penetrated the cyst wall and then oxidant species produced by TiO2/UV destroyed both cell wall and intracellular structure. In the case of G. intestinalis inactivation (disinfection) performance of TiO2/UV system reached 52.5% only after 25 min illumination and total parasite disinfection was achieved after 30 min illumination. However, silver loaded TiO2 seemed to be more effective as this loading provided better catalytic action as well as additional antimicrobial properties. Cell viability tests showed that parasite cysts, their walls in particular, were irreversibly damaged and cysts did not re-grow. Nevertheless the studied system seemed to be ineffective for the inactivation of A. castellani. Inactivation percentages of TiO2/UV and Ag-TiO2/UV systems were far lower than that of UV alone, being 50.1% and 46.1%, respectively.     

Reactions of oxygen radicals with the quinone ring of coenzyme Q.

Coenzyme Q, besides its role in electron transfer reactions, may act as a radical scavenger. The effect of oxygen radicals produced by ultrasonic irradiation on the quinone ring was investigated. Aqueous solutions of a Q homologue, completely lacking the side chain, were irradiated and the modifications were spectrophotometrically followed. The experimental results show that both degradation and reduction of the benzoquinone ring took place when the irradiation was performed in water. Data obtained when ultrasonic irradiation was carried out in the presence of OH. scavengers, as formate, organic and inorganic buffers, suggest: a) the responsible species for most the ubiquinol generated by sonication appeared to be the superoxide radical b) addition reactions of OH. radicals with the aromatic ring led probably to the degradation of Coenzyme Q molecules.     

Ultrasound-assisted enzymatic transesterification of methyl benzoate and glycerol to 1-glyceryl benzoate in organic solvent

The aim of this work is to report the enzymatic transesterification production of 1-glyceryl benzoate under ultrasound irradiation, using a commercial immobilized lipase, Novozym 435. Firstly, a preliminary evaluation was carried out at 2, 4 and 6h, at constant temperature of 50 °C, methyl benzoate to glycerol molar ratio of 1:1 and 5.5 wt% of enzyme concentration. After analyzing the results obtained, the experimental design technique was used to evaluate the effects of temperature, substrates molar ratio, enzyme concentration, solvent volume and ultrasonic power on the 1-glyceryl benzoate production. The highest conversion, around 16%, was obtained at 65 °C, 1:1 of methyl benzoate to glycerol molar ratio, 15 wt% of enzyme concentration, 7 mL of solvent and 40% ultrasonic power in 4h of reaction. A preliminary kinetic experiment carried out varying the enzyme concentration (15 and 20 wt%) keeping fixed the temperature at 35 °C, 1:1 of substrates molar ratio, 3 mL of solvent and 40% of maximum ultrasonic power led to lower (around 15% after 12 h of reaction) conversions compared to that achieved in the experimental design.     

Additional effects of silver nanoparticles on bactericidal efficiency depend on calcination temperature and dip-coating speed

There is an increasing interest in the application of photocatalytic properties for disinfection of surfaces, air, and water. Titanium dioxide is widely used as a photocatalyst, and the addition of silver reportedly enhances its bactericidal action. However, the synergy of silver nanoparticles and TiO(2) is not well understood. The photocatalytic elimination of Bacillus atrophaeus was examined under different calcination temperatures, dip-coating speeds, and ratios of TiO(2), SiO(2), and Ag to identify optimal production conditions for the production of TiO(2)- and/or TiO(2)/Ag-coated glass for surface disinfection. Photocatalytic disinfection of pure TiO(2) or TiO(2) plus Ag nanoparticles was dependent primarily on the calcination temperature. The antibacterial activity of TiO(2) films was optimal with a high dip-coating speed and high calcination temperature (600°C). Maximal bacterial inactivation using TiO(2)/Ag-coated glass was also observed following high-speed dip coating but with a low calcination temperature (250°C). Scanning electron microscopy (SEM) showed that the Ag nanoparticles combined together at a high calcination temperature, leading to decreased antibacterial activity of TiO(2)/Ag films due to a smaller surface area of Ag nanoparticles. The presence of Ag enhanced the photocatalytic inactivation rate of TiO(2), producing a more pronounced effect with increasing levels of catalyst loading.     

超声参数对白术多糖抗氧化活性的影响

采用热水浸提和超声辅助法提取白术多糖,研究白术多糖的总还原能力以及对超氧阴离子(O2-·)、羟基自由基(·OH)、1,1-二苯基-2-苦苯肼自由基(DPPH·)的清除作用.结果表明:超声频率80 kHz,功率220 W时,白术多糖提取率达到最大(8.017％),明显高于热水浸提法所得(6.832％).白术多糖对·OH和DPPH·具有较强的清除作用,随着质量浓度的升高,·OH清除率呈上升趋势.超声功率和频率均对白术多糖的抗氧化活性有影响且超声功率的影响较大.随着超声功率的提高,白术多糖清除·OH的能力增强,但对O2-·的清除能力影响较弱.     

Direct and Rapid Analysis of the Adhesion of Bacteria to Solid Surfaces: Interaction of Fluorescently Labeled Rhodococcus Strain GIN-1 (NCIMB 40340) Cells with Titanium-Rich Particles

A fluorimetric assay which enables direct and accurate analysis of the adhesion of bacteria to solid particles was developed. The assay is based on labeling of the bacteria with fluorescamine, which reacts with primary amino groups on the cell surface to yield a yellow fluorescence that is easily detectable by both fluorescence microscopy and spectrofluorimetry. As an example, fluorescent labeling of Rhodococcus strain GIN-1 (NCIMB 40340) cells enabled the detection and quantitative determination of their adsorption to TiO(inf2) and coal fly ash particles. Exposure of the cells to 10% acetone during the labeling reaction affected neither their viability nor their ability to adhere to these particles. Only a small fraction (^sim2%) of the total cell protein was labeled by fluorescamine upon staining of intact bacterial cells, which may indicate preferential labeling of certain proteins. Specificity studies carried out with the fluorescence assay confirmed previous findings that Rhodococcus strain GIN-1 cells possess high affinities for TiO(inf2), ZnO, and coal fly ash and low affinities for other metal oxides. In principle, the newly developed fluorimetric assay may be used for determination of cell adhesion to any solid matrix by either microscopic examination or epifluorescence measurements. In the present work, the adhesion of several other microorganisms to TiO(inf2) particles was tested as well, but their ability to adhere to these particles was significantly lower than that of Rhodococcus strain GIN-1 cells.     

响应面优化超声辅助法提取拐枣种子中二氢杨梅素的工艺

二氢杨梅素和杨梅素是拐枣种子中的重要成分。通过超声辅助法从拐枣种子中提取二氢杨梅素，采用单因素法考察乙醇体积分数、超声辐照功率、提取温度、液料比和超声辐照时间影响参数的基础上，并选用Box-Behnken响应面设计法分析建立了超声辐照功率、超声辐照时间和液料比的二次多项式模型，优化提取工艺。结果得到超声辅助法提取拐枣种子中二氢杨梅素的最佳工艺参数为：乙醇体积分数为60%、超声辐照功率140 W、超声辐照时间30 min、液料比20.5 mL·g^-1，提取温度40℃。在此最佳条件下，二氢杨梅素得率为2.14±0.09 mg·g^-1。本提取方法简单快速，效率高，有利于拐枣资源的综合加工利用。     

Infectious DNA from coliphage T1

Summary Infectious DNA from phage T1 was inactivated by UV-light (2,537 Å). No effect of irradiation on the kinetics of the assay in a spheroplast system could be observed. UV-damaged molecules compete with unirradiated DNA for the infection. Infectious T1-DNA is subject to host-cell reactivation of UV-damage, the amount of which depends on the physiological conditions of the spheroplasts. Though UV-radiosensitivity of T1 particles is not influenced by the presence of the radical scavenging compound cysteamine, infectious DNA can be protected effectively by this chemical (0.01M) against UV-damage when HCR-negative spheroplasts are used for the assay. Incorporation of 5-bromouracil radiosensitizes infectious T1-DNA in the presence and absence of HCR. This effect can be eliminated when the DNA is irradiated in the presence of cysteamine. The mechanism of radioprotection is discussed.     

Microbial denitrification by immobilized bacteria Pseudomonas denitrificans stimulated by constant electric field

A comparative study of microbial reduction of nitrate ions by immobilized Pseudomonas denitrificans cells in the presence and in the absence of a constant electric field was carried out. The experiments were carried out in a continuous stirred tank reactor at a broad range of dilution rates. The cathode potential was maintained constant around the standard potential value of the nitrate/nitrite redox couple. The residual outlet concentrations of nitrate, when the electric field was applied, were much lower than those obtained without the electric field, all other conditions being equal. It was demonstrated that the electric field had a significant effect on the nitrate reduction rate allowing the use of much higher inlet flow rates compared to the flow rate in the absence of an electric field.     

The use of the pulse radiolysis method in the study of rapid processes in suspensions of Escherichia coli

It was proved that the method of pulse radiolysis can be used to study the processes rapidly occurring in a microorganism suspension. It was shown that optically active particles formed after irradiation in an E. coli cell were involved into the first-(pseudo-first)-order reaction the rate constant being approximately 30 s-1. The presence of oxygen did not influence the kinetics of disappearance of these particles.     

Controllable preparation of Nano-MgO and investigation of its bactericidal properties

Samples of nano-MgO with varying particle sizes were prepared by four different methods using Mg(NO3)2.6H2O, Na2CO3, urea and ammonia as raw materials and characterized by X-ray diffraction (XRD), transmission electron microscopy (TEM), low temperature N2 adsorption-desorption measurements and FT-IR spectroscopy. Bactericidal experiments with Bacillus subtilis var. niger and Staphylococcus aureus were carried out using as-synthesized nano-MgO samples and the bactericidal mechanism was also investigated. The results showed that the bactericidal efficacy of nano-MgO increases with decreasing particle size. The bactericidal efficacy of the samples was compared with that of TiO2, a common photoactive bactericidal material. The nano-MgO has better bactericidal activity, both when used directly and as an additive in an interior wall paint. Furthermore, nano-MgO is active even in the absence of irradiation.     

Ultrasonication enhanced hydrolytic activity of lipase in water/isooctane two-phase systems

The hydrolysis of olive oil catalyzed by Chromobacterium viscosum lipase (EC 3.1.1.3) in a water/isooctane two-phase system was carried out both under ultrasound and conventional stirring. The maximum activity of lipase in the ultrasonicated system was 1.75 times higher than that in the stirred system. The lipase activity was dependent on ultrasonic power and volume ratio of isooctane to water. The optimum reaction temperature in both systems was around 25°C. The stability of lipase at 25°C in the ultrasonicated system decreased more rapidly than that in the stirred system. In the presence of exogenous oleic acid, however the half-life of lipase in the ultrasonicated system was improved to a value, which was respectively half and twice of that in stirred systems with and without oleic acid. The maximum reaction rate (V_max) was increased by ultrasonication whereas the Michaelis constant (K_m) remained unaltered.     

Free radical induced inactivation of creatine kinase: sites of interaction, protection, and recovery

The study aims at a clarification of the oxidative damage of creatine kinase isoenzymes by X-ray-induced water radiolysis. The radical species generated by this method (under appropriate conditions) are similar to those discussed in the context of mitochondrial energy metabolism. The decay of the enzyme activity is accompanied by a strong decrease of the number of accessible SH groups and by a reduction of the endogenous tryptophan fluorescence. Free radical effects are diminished if irradiation is carried out in the presence of 2-mercaptoethanol. Partial recovery of the activity (repair) is observed if 2-mercaptoethanol is added after irradiation. The experiments suggest a twofold importance of thiol reagents (RSH): to reduce the concentration of free radicals by scavenger reactions and to modify the inactivation mechanism in such a way that efficient repair of enzyme damage may be achieved. Cysteine 282 of MM-CK (Cys-278 in the case of Mi-CK) seems to play a crucial role in this respect. Blockage of the SH group of cysteine 282 by oxidized glutathione effectively protects the enzyme against inactivation by NO(*)(2) radicals. In the absence of nitrogen dioxide and of thiol reagents, however, inactivation seems to proceed via a less specific mechanism involving additional targets of the enzyme.     

Prooxidative activities of tea catechins in the presence of Cu2+

The ability of various tea catechins to generate H2O2 and the hydroxyl radical in the presence of the Cu2+ ion was investigated and compared with the effect of iron ions. The presence of Cu2+ accelerated the generation of H2O2 by EGC, while EGCg with Cu2+ generated a little H2O2. The presence of iron ions inhibited the generation of H2O2 by EGC. EGC and EC with Cu2+ generated the hydroxyl radical, while EGCg and ECg with Cu2+ did not. The fact that EGCg showed less prooxidative activity than EGC can be explained by the chelating ability of catechin gallates to metal ions under the experimental conditions.     

Defect in the split proteins of 30-S ribosomal subunits and under-methylation of 16-S ribosomal RNA in a polyamine-requiring mutant of Escherichia coli grown in the absence of polyamines.

Polyphenylalanine synthesis was carried out with Escherichia coli Q13 50-S ribosomal subunits and reconstituted 30-S particles containing different combinations of 23-S core particles and 30-S subunit split proteins obtained from a polyamine-requiring mutant of E. coli during its growth in the presence or absence of putrescine. It was concluded that the defect in the amount of some kinds of 30-S subunit split proteins was responsible for the decrease of polypeptide synthesis in a polyamine-requiring mutant of E. coli grown in the absence of polyamines. The methylation of 16-S RNA during growth in the absence of putrescine was decreased, while the degree of methylation of 23-S RNA did not change significantly. The decrease in methylation of 16-S RNA in the absence of putrescine was due mainly to a decrease of methylation of adenine. The relationship between the decrease of polypeptide synthetic activity of 30-S ribosomal subunits obtained from a polyamine-requiring mutant of E. coli grown in the absence of polyamines and the decrease of methylation of 16-S RNA is discussed.     

Stoichiometric and kinetic characterisation of Nitrobacter in mixed culture by decoupling the growth and energy generation processes

The growth, maintenance and lysis processes of Nitrobacter were characterised. A Nitrobacter culture was enriched in a sequencing batch reactor (SBR). Fluorescent in situ hybridisation showed that Nitrobacter constituted 73% of the bacterial population. Batch tests were carried out to measure the oxygen uptake rate and/or nitrite consumption rate when both nitrite and CO2 were in excess, and in the absence of either of these two substrates. The results obtained, along with the SBR performance data, allowed the determination of the maintenance coefficient and in situ cell lysis rate of Nitrobacter. Nitrobacter spends a significant amount of energy for maintenance, which varies considerably with the specific growth rate. At maximum growth, Nitrobacter consume nitrite at a rate of 0.042 mgN/mgCOD(biomass) . h for maintenance purposes, which increases more than threefold to 0.143 mgN/mgCOD(biomass) . h in the absence of growth. In the SBR, where Nitrobacter grew at 40% of its maximum growth rate, a maintenance coefficient of 0.113 mgN/mgCOD . h was found, resulting in 42% of the total amount of nitrite being consumed for maintenance. The above three maintenance coefficient values obtained at different growth rates appear to support the maintenance model proposed in Pirt (1982). The in situ lysis rate of Nitrobacter was determined to be 0.07/day under aerobic conditions at 22 degrees C and pH 7.3. Further, the maximum specific growth rate of Nitrobacter was estimated to be 0.02/h (0.48/day). The affinity constant of Nitrobacter with respect to nitrite was determined to be 1.50 mgNO2(-)-N/L, independent of the presence or absence of CO2.