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The antioxidative role of cerium was investigated in the liver of silver crucian carp injected with lead. The fish were intraperitoneally injected with 10, 20, or 30 mg/kg wet weight PbCl(2). After a 14-day period of incubation, 35 animals were injected with a solution of 1.5 mg/kg wet weight CeCl(3). After 42 days, the wet weight and the liver weight of the fish were weighed, and the oxidative stress of the fish liver was estimated by assaying lipid peroxide, superoxide dismutase, catalase, ascorbate peroxidase, glutathione peroxidase, glutathione, ascorbic acid, and reactive oxygen species (ROS). The results show that Ce(3+) could decrease ROS accumulation, relieve the inhibition of the activities of the antioxidant enzyme and the reduction of antioxidants in fish liver caused by Pb(2+), and decrease the enhancement of hepatosomatic index of fish under various Pb(2+) dosages.

The activity of alanine aminotransferase (ALT; E.C. 2.6.1.2) is often changed upon inflammatory responses in animals. Rare earths was shown to provoke various inflammatory responses both in rats and mice; however, the molecular mechanism by which rare earths exert its toxicity has not been completely understood, especially, we know little about the mechanism of the interaction between CeCl(3) and ALT. In this report, we investigated the mechanisms of CeCl(3) on ALT activity in vivo and in vitro. Our results showed that Ce(3+) could significantly activate ALT in vivo and in vitro; the kinetics constant (Km) and Vmax were 0.018 microM and 1,380 unit mg(-1) protein min(-1), respectively, at a low concentration of Ce(3+), and 0.027 microM and 624 unit mg(-1) protein min(-1), respectively, at a high concentration of Ce(3+). By UV absorption and fluorescence spectroscopy assays, the Ce(3+) was determined to be directly bound to ALT; the binding site of Ce(3+) to ALT was 1.72, and the binding constants of the binding site were 4.82 x 10(8) and 9.05 x 10(7) L mol(-1). Based on the analysis of the circular dichroism spectra, it was concluded that the binding of Ce(3+) altered the secondary structure of ALT, suggesting that the observed enhancement of ALT activity was caused by a subtle structural change in the active site through the formation of the complex with Ce(3+).

This study was performed with the objective of assessing the antioxidant response of the lung of mice to different rare earths. LaCl(3), CeCl(3), and NdCl(3) at a higher dose of 20 mg/kg body weight were injected into the nasal cavity of ICR mice for consecutive 14 days, respectively. The increase of pulmonary lipids peroxide produced by Ln suggested an oxidative attack that was activated by a reduction of antioxidative defense mechanisms as measured by analyzing the activities of superoxide dismutase, catalase, ascorbate peroxidase, and total antioxidant capacity, as well as antioxidant levels such as glutathione and ascorbic acid, which were greatest in Ce(3+) treatment, medium in Nd(3+), and least in La(3+). It implied that the antioxidative responses of lung may be involved in 4f shell and alternable valence properties of Ln-induced lung toxicity.

Although it is known that nano-TiO2or other nanoparticles can induce liver toxicities, the mechanisms and the molecular pathogenesis are still unclear. In this study, nano-anatase TiO2(5 nm) was injected into the abdominal cavity of ICR mice for consecutive 14 days, and the inflammatory responses of liver of mice was investigated. The results showed the obvious titanium accumulation in liver DNA, histopathological changes and hepatocytes apoptosis of mice liver, and the liver function damaged by higher doses nano-anatase TiO2. The real-time quantitative RT-PCR and ELISA analyses showed that nano-anatase TiO2can significantly alter the mRNA and protein expressions of several inflammatory cytokines, including nucleic factor-κB, macrophage migration inhibitory factor, tumor necrosis factor-α, interleukin-6, interleukin-1β, cross-reaction protein, interleukin-4, and interleukin-10. Our results also implied that the inflammatory responses and liver injury may be involved in nano-anatase TiO2-induced liver toxicity.

Previous research on the biological and toxic effects of nano-TiO(2) particles on animals only limit to a single dose. However, the toxicity caused by single dose nano-TiO(2) does not truly represent ecological and health effects of nano-TiO(2) retained in the environment. In order to further evaluate the toxicity of nano-TiO(2) particles, nano-anatase TiO(2) (5 nm) was injected into the abdominal cavity of ICR mice everyday for 14 days and the coefficients of organs and serum biochemical parameters were investigated. The results showed that, with increasing doses of nano-anatase TiO(2), the coefficients of liver, kidney, and spleen increased gradually, while the coefficients of lung and brain decreased gradually, and the coefficient of heart had little change. The order of the titanium accumulation in the organs was liver > kidneys > spleen > lung > brain > heart. The serum biochemical parameters with lower dose of nano-anatase TiO(2) showed little difference compared with the control mice, while with higher dose of nano-anatase TiO(2), the indicators of liver function, such as alkaline phosphatase, alanine aminotransferase, leucine acid peptide, pseudocholinesterase, total protein, and albumin level, were enhanced significantly; the indicators of kidney function, such as uric acid and blood urea nitrogen, were decreased; the activities of aspartate aminotransferase, creatine kinase, lactate dehydrogenase, and alpha-hydroxybutyrate dehydrogenase, indicator of the myocardium function, were increased. The contents of triglycerides, glucose, and high-density lipoprotein cholesterol were significantly elevated. Taken together, nano-anatase TiO(2) in higher dose caused serious damage to the liver, kidney, and myocardium of mice and disturbed the balance of blood sugar and lipid in mice. The accumulation of titanium in the organs might be closely related to the coefficients of organs and the inflammatory responses of mice.

Characterized by a photo-catalysis property, nano-anatase TiO(2) is closely related to photosynthesis of spinach. It could not only improve light absorbance, transformation from light energy to electron energy and active chemical energy, but also promote the activity of Rubiso activase of spinach. However, the relation between the activity of Rubiso activase and the growth of spinach promoted by nano-anatase TiO(2) treatment remains largely unclear. In this study, we find that the amount and the activity of Rubiso activase are obviously increased by nano-anatase TiO(2 )treatment, which led to the great promotion of Rubsico carboxylation and the high rate of photosynthesis, thus improving of spinach growth. The significant enhancement of Rubiso activase activity of nano-anatase TiO(2 )treated spinach is also accompanied by conformational changes as determined by spectroscopic analysis. But bulk TiO(2) effect is not as significant as nano-anatase TiO(2), as the grain size of nano-anatase TiO(2) (5 nm) is much smaller than that of bulk TiO(2), which entered spinach cell more easily.

The subject of acute injury, apoptosis and canceration of animals induced by heavy metal ions has been one of the hotspots studied worldwide. However, the exact molecular mechanism of Cd-induced carcinogenicity remains largely unclear, and how to relieve the toxicity in vivo has rarely been reported. For this paper, we have investigated the mechanism of Cd2+ on DNA cleavage and Ca2+ on DNA repair in the liver of silver crucian carp (Carassius auratus gibelio) by agarose gel electrophoresis methods and by estimating biochemical indexes. Our results show that Cd2+ induces the classical laddering degradation of DNA in vivo and that DNA cleavage is repaired after injection with Ca2+ under various Cd2+ concentrations. DNA cleavage caused by Cd2+ is due to the activation of deoxyribonuclease (DNase) and the accumulation of reactive oxygen species (ROS). Furthermore, Cd2+ destroys the antioxidant system, which diminishes the activities of superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD), causing an increase of the lipid peroxidation (LPO) level, respectively. However, after the liver is injected with Ca2+ under various Cd2+ concentrations, the DNase activity, the ROS generating rate and the LPO level are obviously reduced, the activities of SOD, CAT, and POD are greatly increased. At the same time, ROS production and removal recoves its balance. The results show that Ca2+ can relieve the toxicity of Cd2+ in silver crucian carp.

The improvement of spinach growth is proved to relate to N2 fixation by nano-anatase TiO2 in this study. The results show that all spinach leaves kept green by nano-anatase TiO2 treatment and all old leaves of control turned yellow white under culture with N-deficient solution. And the fresh weight, dry weight, and contents of total nitrogen, NH4(+), chlorophyll, and protein of spinach by nano-anatase TiO2 treatment presented obvious enhancement compared with control. Whereas the improvements of yield of spinach were not as good as nano-anatase TiO2 treatment under N-deficient condition, confirming that nano-anatase TiO2 on exposure to sunlight could chemisorb N2 directly or reduce N2 to NH3 in the spinach leaves, transforming into organic nitrogen and improving the growth of spinach. Bulk TiO2 effect, however, was not as significant as nano-anatase TiO2. A possible metabolism of the function of nano-anatase TiO2 reducing N2 to NH3 was discussed.

The effects of nano-anatase TiO2 on light absorption, distribution, and conversion, and photoreduction activities of spinach chloroplast were studied by spectroscopy. Several effects of nano-anatase TiO2 were observed: (1) the absorption peak intensity of the chloroplast was obviously increased in red and blue region, the ratio of the Soret band and Q band was higher than that of the control; (2) the great enhancement of fluorescence quantum yield near 680 nm of the chloroplast was observed, the quantum yield under excitation wavelength of 480 nm was higher than the excitation wavelength of 440 nm; (3) the excitation peak intensity near 440 and 480 nm of the chloroplast significantly rose under emission wavelength of 680 nm, and F 480 / F 440 ratio was reduced; (4) when emission wavelength was at 720 nm, the excitation peaks near 650 and 680 nm were obviously raised, and F 650 / F 680 ratio rose; (5) the rate of whole chain electron transport, photochemical activities of PSII DCPIP photoreduction and oxygen evolution were greatly improved, but the photoreduction activities of PSI were a little changed. Together, the studies of the experiments showed that nano-anatase TiO2 could increase absorption of light on spinach chloroplast and promote excitation energy to be absorbed by LHCII and transferred to PSII and improve excitation energy from PSI to be transferred to PSII, thus, promote the conversion from light energy to electron energy and accelerate electron transport, water photolysis, and oxygen evolution.

A direct interaction of lead (Pb) with kidney DNA from silver crucian carp (Carassius auratus gibelio) has been systematically studied in vitro using multi-techniques, including UV-vis absorption, extended X-ray absorption fine structure, vacuum ultraviolet circular dichroism spectral methods and gel electrophoresis. We find that Pb is bound with four oxygen or nitrogen atoms of DNA in its fresh shell at the distances of 2.64 A. Additionally, Pb may bind to the oxygen atom of nucleic acid or nitrogen atom of base pairs of DNA in that obvious changes in the secondary structure of DNA are found after Pb treatment. However, this direct interaction of Pb with DNA do not cause DNA cleavage in vitro.

The mechanism of Cd2+ on the DNA cleavage and Ce3+ on the DNA repair in the kidney of silver crucian carp (Carassius auratus gibelio) is investigated by agarose gel electrophoresis methods and assaying biochemical indexes. It proves that Cd2+ induces the classical laddering degradation of DNA in vivo, but DNA cleavage is repaired after injecting with a low Ce3+ concentration under various Cd2+ concentrations. The DNA cleavage caused by Cd2+ is the result of the activation of deoxyribonuclease (DNase) and accumulation of reactive oxygen species (ROS), and Cd2+ destroys the antioxidant system, which diminishes the activities of superoxide dismutase, catalase, and peroxidase, and the increase of the lipid peroxidation (LPO) level. However, Ce3+ could inhibit activation of Cd2+ on DNase activity, relieve inhibition of Cd2+ on activities of the antioxidant enzyme, and diminish ROS accumulation. The results show that Ce3+ could relieve the toxicity of Cd2+ to silver crucian carp.

Having a photocatalyzed characteristic, our previous research had proved that nano-anatase TiO2 is closely related to the photosynthesis of spinach. It could not only improve the light absorbance and the transformation from light energy to electron energy and to active chemical energy but also promote carbon dioxide (CO2) assimilation of spinach. However, the mechanism of carbon reaction promoted by nano-anatase TiO2 remains largely unclear. By electrophoresis and Western blot methods, the results of the experiments proved that Rubisco from the nano-anatase TiO2-treated spinach during the extraction procedure of Rubisco was found to consist of Rubisco and a heavier molecular-mass protein (about 1,200 kDa) comprising both Rubisco and Rubisco activase. The Rubisco carboxylase activity was 2.67 times that of Rubisco from the control and it could hydrolyze ATP in the same manner as Rubisco activase. The total sulfhydryl groups and available sulfhydryl groups of the Rubisco were 32 -SH and 21 -SH per mole of enzyme more than those of the Rubisco purified from the control, respectively. The circular dichroism spectra showed that the secondary structure of Rubisco from the nano-anatase TiO2-treated spinach was very different from Rubisco of the control. It suggested that the mechanism of nano-anatase TiO2 activating Rubisco of spinach was that the complex of Rubsico and Rubisco activase was induced in spinach, which promoted Rubsico carboxylation and increased the rate of photosynthetic carbon reaction.

Previous research showed that nano-TiO2 could significantly promote photosynthesis and greatly improve growth of spinach, but we also speculated that an increase of spinach growth by nano-TiO2 treatment might be closely related to the change of nitrogen metabolism. The effects of nanoanatase TiO2 on the nitrogen metabolism of growing spinach were studied by treating them with nano-anatase TiO2. The results showed that nano-anatase TiO2 treatment could obviously increase the activities of nitrate reductase, glutamate dehydrogenase, glutamine synthase, and glutamic-pyruvic transaminase during the growing stage. Nano-anatase TiO2 treatment could also promote spinach to absorb nitrate, accelerate inorganic nitrogen (such as NO3--N and NH4+-N) to be translated into organic nitrogen (such as protein and chlorophyll), and enhance the fresh weight and dry weights.

Cadmium is one of the most toxic heavy metals and is known to accumulate in freshwater food chains. The underlying mechanism for its genotoxicity has not been investigated for any freshwater fish. It has, however, been suggested that cadmium-induced carcinogenesis might involve either direct or indirect interaction of Cd2+ with DNA. The interaction between Cd2+ and DNA from the kidney of the silver crucian carp (Carassius auratus gibelio) in vitro and in vivo is investigated by spectrophotometric methods and agarose gel electrophoresis methods. Cd2+ could insert into DNA basepairs, bind to nucleic acid, and result in notable hypochromicities. The analysis of agarose gel electrophoresis proves that Cd2+ at different concentrations does not cause DNA cleavage in vitro; however, kidneys display the classical laddering degradation of DNA in vivo, which is the result of the promotion of deoxyribonuclease activity or inhibition of superoxide dismutase and catalyse activity and the accumulation of reactive oxygen species caused by Cd2+ ions in vivo.

Chloroplasts and chlorophylls were isolated from the leaves of Dicranopteris linearis, a natural perennial fern sampled at rare earth element (REE) mining areas in the South-Jiangxi region (southern China). The inductively coupled plasma-mass spectrometry (ICP-MS) results indicated that REEs were present in the chloroplasts and chlorophylls of D. linearis. The in vivo coordination environment of light REE (lanthanum) or heavy REE (yttrium) ions in D. linearis chlorophyll-a was determined by the extended X-ray absorption fine structure (EXAFS). Results revealed that there were eight nitrogen atoms in the first coordination shell of the lanthanum atom, whereas there were four nitrogen atoms in the first coordination shell of yttrium. It was postulated that the lanthanum-chlorophyll-a complex might have a double-layer sandwich-like structure, but yttrium-binding chlorophyll-a might be in a single-layer form. Because the content of REE-binding chlorophylls in D. linearis chlorophylls was very low, it is impossible to obtain structural characteristics of REE-binding chlorophylls by direct analysis of the Fourier transform infrared (FTIR) and ultraviolet (UV)-visible spectra of D. linearis chlorophylls. In order to acquire more structural information of REE-binding chlorophyll-a in D. linearis, lanthanum - and yttrium-chlorophyll-a complexes were in vitro synthesized in acetone solution. Element analyses and EXAFS results indicated that REE ions (lanthanum or yttrium) of REE-chlorophyll-a possessed the same coordination environment whether in vivo or in vitro. The FTIR spectra of the REE-chlorophyll-a complexes indicated that REEs were bound to the porphyrin rings of chlorophylls. UV-visible results showed that the intensity ratios of Soret to the Q-band of REE-chlorophyll-a complexes were higher than those of standard chlorophyll-a and pheophytin-a, indicating that REE-chlorophyll-a might have a much stronger ability to absorb the ultraviolet light. The MCD spectrum in the Soret band region of lanthanum-chlorophyll-a showed a special peak, but yttrium-chlorophyll-a did not have this special peak, corresponding well to their double-layer and single-layer structure, respectively.

The interactions between Hg2+, Ce3+, and the mixture of Ce3+ and Hg2+, and DNA from fish intestine in vitro were investigated by using absorption spectrum and fluorescence emission spectrum. The ultraviolet absorption spectra indicated that the addition of Hg2+, Ce3+, and the mixture of Ce3+ and Hg2+ to DNA generated an obviously hypochromic effect. Meanwhile, the peak of DNA at 205.2 nm blue-shifted and at 258.2 nm red-shifted. The size of the hypochromic effect and the peak shift of DNA by metal ion treatments was Hg2+>Hg2++Ce3+>Ce3+. The fluorescence emission spectra showed that with the addition of Hg2+, Ce3+, and the mixture of Ce3+ and Hg2+ the emission peak at about 416.2 nm of DNA did not obviously change, but the intensity reduced gradually and the sequence was Hg2+>Hg2++ Ce3+>Ce3+. Hg2+, Ce3+, and the mixture of Ce3+ and Hg2+ had 1.12, 0.19, and 0.41 binding sites to DNA, respectively; the fluorescence quenching of DNA caused by the metal ions all attributed to static quenching. The binding constants (KA) of binding sites were 8.98 x 104 L/mol and 1.02 x 104 L/mol, 5.12 x 104 L/mol and 1.10 x 103 L/mol, 6.66 x 104 L/mol and 2.36 x 103 L/mol, respectively. The results showed that Ce3+ could relieve the destruction of Hg2+ on the DNA structure.

The effects of nano-TiO2 (rutile) on the photochemical reaction of chloroplasts of spinach were studied. The results showed that when spinach was treated with 0.25% nano-TiO2, the Hill reaction, such as the reduction rate of FeCy, and the rate of evolution oxygen of chloroplasts was accelerated and noncyclic photophosphorylation (nc-PSP) activity of chloroplasts was higher than cyclic photophosphorylation (c-PSP) activity, the chloroplast coupling was improved and activities of Mg2+-ATPase and chloroplast coupling factor I (CF1)-ATPase on the thylakoid membranes were obviously activated. It suggested that photosynthesis promoted by nano-TiO2 might be related to activation of photochemical reaction of chloroplasts of spinach.

The effects of nano-TiO2 (rutile) on the chloroplast aging of spinach under light were studied. The results showed that when the chloroplasts were illuminated for 1, 5, and 10 min with 500 micromol/cm2/min light intensity, respectively, the evolution oxygen rate was rapidly increased; when the chloroplasts were treated for 20, 30, and 40 min with 500 micromol/cm2/min light intensity, respectively, the evolution oxygen rate was gradually decreased. While spinach was treated with 0.25% nano-TiO2, the rate of evolution oxygen of chloroplasts in different illumination times (1, 5, 10, 20, 30, and 40 min) was higher than that of control, and when the illumination time was over 10 min, the reduction of the evolution oxygen rate was lower than that of control. It suggested that nano-TiO2 treatment could protect chloroplasts from aging for long-time illumination. The mechanism researches indicated that nano-TiO2 treatment could significantly increase the activities of superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD), decrease accumulation of reactive oxygen free radicals and the level of malondialdehyde (MDA), and maintain stability of membrane structure of chloroplast under light.

The effects of nano-TiO(2) (rutile) and non-nano-TiO(2) on the germination and growth of naturally aged spinach seeds were studied by measuring the germination rate and the germination and vigor indexes of aged spinach seeds. An increase of these factors was observed at 0.25-4% nano-TiO(2) treatment. During the growth stage, the plant dry weight was increased, as was the chlorophyll formation, the ribulosebisphosphate carboxylase/oxygenase activity, and the photosynthetic rate. The best results were found at 2.5% nano-TiO(2). The effects of non-nano-TiO(2) are not significant. It is shown that the physiological effects are related to the nanometer-size particles, but the mechanism by which nano-TiO(2) improves the growth of spinach seeds still needs further study.

The mechanism of La(3+) effect on chlorophyll (chl) of spinach in solution culture has been studied. The results show that La(3+) can obviously promote growth, increase chlorophyll contents and photosynthetic rate of spinach. La(3+) may substitute Mg(2+) for chlorophyll formation of spinach when there is no Mg(2+) in solution. La(3+) improves significantly PSII formation and enhances electron transport rate of PSII. By ICP-MS and atom absorption spectroscopy methods, it has been revealed that rare earth elements (REEs) can enter chloroplasts and increase Mg(2+)-chl contents; and REEs bind to chlorophyll and also form REE-chl. REE-chl is about 72% in total chlorophyll with La(3+) treatment and without Mg(2+)in solution. By UV-Vis, FT-IR and extended X-ray absorption fine structure spectroscopy (EXAFS) methods, it has been found that La(3+) coordinates with nitrogen of porphyrin rings with the average La-N bond length of 0.253 nm.