Increased selenium concentration in the synthesis of CdSe magic-sized quantum dots affects how the brain responds to oxidative stress.

CdSe magic-sized quantum dots (MSQDs) have been widely used as fluorescent probes in biological systems due to their excellent optical properties with a broader fluorescence spectrum and stable luminescence in biological media. However, they can be cytotoxic and alter the redox balance depending on the amounts of Cd2+ adsorbed on their surface. Thus, the present study aimed to evaluate whether increases in selenium concentration in the synthesis of CdSe-MSQDs decrease the oxidative stress caused by Cd2+ -based quantum dots. CdSe-MSQDs synthesized with different concentrations of selenium were investigated against oxidative stress in the brain of chicken embryos by examining total antioxidant capacity, lipid peroxidation, thiol, and glutathione contents, as well as the activities of glutathione peroxidase, superoxide dismutase (SOD), catalase (CAT), and glutathione reductase. In addition, the vascularization of the chorioallantoic membrane (CAM) analysis was performed. Higher selenium concentrations alter the surface defect levels (decrease free Cd2+ ) and controlled the oxidative effects of CdSe-MSQDs by reducing the lipid peroxidation, restoring the glutathione defense system and the antioxidant enzymes SOD and CAT, and maintaining the vascular density of the CAM. The current findings reinforce the study of the effects of the presence of Cd2+ ions on the surface of quantum dots, changing toxicity, and aiming interesting strategies of nanomaterials in biological systems.

Effects of Photobiomodulation on Daphnia magna Straus and their Sensitivity to Toxicant.

This paper deals with the effect of photobiomodulation (PBM) on Daphnia magna S. and their sensitivity to cadmium sulfate, a known high toxic pollutant. In a first series of experiments, the effect of different He-Ne laser fluences irradiation (range 0.9-4300 mJ cm-2 ) on the fertility of both parent and filial generations (F1-F3) of the crustacean was studied. It was found that PBM in some cases significantly influenced the fertility of both irradiated crustaceans and their nonirradiated offspring. By selecting two fluences (9 ± 2 mJ cm-2 reducing fertility and 4.3 ± 0.9 J cm-2 increasing it), the effect of these on toxicity of cadmium sulfate was evaluated. These experiments have shown that prior irradiation with low-intensity light of a helium-neon laser with 632.8 nm wavelength can change the sensitivity of aquatic organisms to toxin cadmium sulfate. The degree and direction of changes depend on the toxicant concentration and the irradiation dose.

Inhibition of biofilm formation by Cd2+ on Bacillus subtilis 1JN2 depressed its biocontrol efficiency against Ralstonia wilt on tomato.

Bacillus subtilis 1JN2 can serve as an effective biocontrol agent against Ralstonia wilt on tomato, but the efficiency of control depends on the levels of heavy metals in the rhizosphere soil. Here, we investigated how the heavy metal Cd2+ affects the biocontrol efficacy of B.subtilis 1JN2 on Ralstonia wilt. We found that low Cd2+ content of 2 mM or lower had no effects on the biofilm formation of 1JN2, while media containing 3 mM or higher Cd2+ levels inhibited biofilm formation. Interestingly, high concentration of Cd2+ (5 mM) showed inhibition of B.subtilis 1JN2 cell growth. We next tested the effects of Cd2+ on the colonization of 1JN2 by supplementing artificial Cd2+ in the tomato rhizosphere in a greenhouse setting. We found that 3 mM Cd2+ in the tomato rhizosphere inhibited the colonization of B.subtilis 1JN2, Only 103 CFU/mL 1JN2 was detected one week post treated with 107 CFU/mL but 105 CFU/mL could be detected without Cd2+ in the soil. The presence of Cd2+ had no effect on the colonization of Ralstonia solanacearum on tomato, but the biocontrol efficacy against Ralstonia wilt by 1JN2 decreased 54.2% when the soil contained 3 mM Cd2+ compared to the control without Cd2+. Taken together, we found that the failure of biofilm formation of Bacillus subtilis 1JN2 that affected by Cd2+ lead to the decrease of its biocontrol efficacy against Ralstonia wilt on tomato.

Down-top nanofabrication of binary (CdO)x (ZnO)1-x nanoparticles and their antibacterial activity.

In the present study, binary oxide (cadmium oxide [CdO])x (zinc oxide [ZnO])1-x nanoparticles (NPs) at different concentrations of precursor in calcination temperature were prepared using thermal treatment technique. Cadmium and zinc nitrates (source of cadmium and zinc) with polyvinylpyrrolidone (capping agent) have been used to prepare (CdO)x (ZnO)1-x NPs samples. The sample was characterized by X-ray diffraction (XRD), scanning electron microscopy, energy-dispersive X-ray (EDX), transmission electron microscopy (TEM), and Fourier transform infrared (FTIR) spectroscopy. XRD patterns analysis revealed that NPs were formed after calcination, which showed a cubic and hexagonal crystalline structure of (CdO)x (ZnO)1-x NPs. The phase analysis using EDX spectroscopy and FTIR spectroscopy confirmed the presence of Cd and Zn as the original compounds of prepared (CdO)x (ZnO)1-x NP samples. The average particle size of the samples increased from 14 to 33 nm as the concentration of precursor increased from x=0.20 to x=0.80, as observed by TEM results. The surface composition and valance state of the prepared product NPs were determined by X-ray photoelectron spectroscopy (XPS) analyses. Diffuse UV-visible reflectance spectra were used to determine the optical band gap through the Kubelka-Munk equation; the energy band gap was found to decrease for CdO from 2.92 to 2.82 eV and for ZnO from 3.22 to 3.11 eV with increasing x value. Additionally, photoluminescence (PL) spectra revealed that the intensity in PL increased with an increase in particle size. In addition, the antibacterial activity of binary oxide NP was carried out in vitro against Escherichia coli ATCC 25922 Gram (-ve), Salmonella choleraesuis ATCC 10708, and Bacillus subtilis UPMC 1175 Gram (+ve). This study indicated that the zone of inhibition of 21 mm has good antibacterial activity toward the Gram-positive B. subtilis UPMC 1175.

Effect of cadmium stress on inductive enzymatic and nonenzymatic responses of ROS and sugar metabolism in multiple shoot cultures of Ashwagandha (Withania somnifera Dunal).

Withania somnifera is one of the most important medicinal plant and is credited with various pharmacological activities. In this study, in vitro multiple shoot cultures were exposed to different concentrations (5-300 µM) of cadmium (Cd) as cadmium sulphate to explore its ability to accumulate the heavy metal ion and its impact on the metabolic status and adaptive responses. The results showed that supplemental exposure to Cd interfered with N, P, and K uptake creating N, P, and K deficiency at higher doses of Cd that also caused stunting of growth, chlorosis, and necrosis. The study showed that in vitro shoots could markedly accumulate Cd in a concentration-dependent manner. Enzymatic activities and isozymic pattern of catalase, ascorbate peroxidase, guaiacol peroxidase, peroxidase, glutathione-S-transferase, glutathione peroxidase, monodehydroascorbate reductase, and dehydroascorbate reductase were altered substantially under Cd exposure. Sugar metabolism was also markedly modulated under Cd stress. Various other parameters including contents of photosynthetic pigments, phenolics, tocopherol, flavonoids, reduced glutathione, nonprotein thiol, ascorbate, and proline displayed major inductive responses reflecting their protective role. The results showed that interplay of enzymatic as well as nonenzymatic responses constituted a system endeavor of tolerance of Cd accumulation and an efficient scavenging strategy of its stress implications.

Cadmium triggers kidney cell apoptosis of purse red common carp (Cyprinus carpio) without caspase-8 activation.

Caspase-8, the essential initiator caspase, is believed to play a pivotal role in death receptor-mediated apoptotic pathway. It also participates in mitochondria-mediated apoptosis via cleavage of proapoptotic Bid in mammals. However, its role in fish remains elusive in Cadmium-induced apoptotic pathway. In this study, we isolated the caspase-8 gene from common carp, one of the most important industrial aquatic animals in China using rapid amplification of cDNA ends (RACE). The deduced amino acid sequence of caspase-8 comprised 475 amino acids, which showed approximately 64.1% identity and 79.8% similarity to zebrafish (Danio rerio) caspase-8, possessed two conserved death effector domains, a large subunit and a small subunit. Phylogenetic analysis demonstrated that caspase-8 formed a clade with zebrafish caspase-8. In kidney, cadmium (Cd) exposure triggered apoptosis and increased caspase-3 and -9 activities, whereas it did not affect caspase-8 activity. Real-time quantitative PCR analysis revealed that caspase-8 transcriptional level was not significantly increased in kidney after exposure to Cd. Using Western blot analysis, no caspase-8 cleaved fragment was detected and no significant alteration of procaspase-8 level was found with the same Cd-treated condition. Moreover, the immunopositive staining was predominantly limited to the cytoplasm of renal tubular epithelial cells and no remarkable changes of immunoreactivities were observed using immunohistochemical detection after Cd treatment. The results reveal that Cd can trigger apoptosis, while it cannot activate caspase-8 in purse red common carp.

Linseed oil based nanocapsules as delivery system for hydrophobic quantum dots.

In the present work, the CdSe/ZnS hydrophobic quantum dots were embedded within the polyelectrolyte nanocapsules. The core of the capsules, which consists of a mixture of the linseed oil with chloroform, was prepared using the spontaneous emulsification technique. The obtained emulsions were stabilized with lecithin and encapsulated using the layer-by-layer (LbL) adsorption of polyelectrolytes. The pair of biocompatible polyelectrolytes was used: the cationic poly-l-lysine hydrobromide (PLL) together with the anionic poly-d-glutamic acid sodium salt. The saturation LbL method, which is based on the stepwise formation of consecutive layers on the initial emulsion without the intermediate rinsing step, was applied to form the capsule shells. Their growth was evidenced by the capsule size and electrophoretic mobility measurements. The emulsion and the capsules were deposited on a mica surface and the deposit topology was examined by the means of atomic force microscopy (AFM). The presence of quantum dots within the oil cores was confirmed by recording the fluorescent spectra of the samples containing CdSe/ZnS. In order to evaluate cytotoxicity of the capsules, their influence on the viability of mouse embryonic fibroblasts was examined using the MTT test, followed by optical-microscope observation of morphology of the cells after hematoxylin-eosin staining.

In vivo monitoring of distributional transport kinetics and extravasation of quantum dots in living rat liver.

Although the unique optical properties of surface-modified quantum dots (QDs) have attracted wide interest in molecular biology and bioengineering, there are very few reports of their in vivo biodistribution, due to a lack of analytical techniques for characterizing the dynamic variation of QDs in living animals. In this study, we used an in vivo online monitoring system and a batch-wise elemental analytical method to investigate the biodistribution/extravasation of various surface-modified CdTeSe/ZnS (QDs) in rat liver. It is found that the surface modification dictated not only the blood retention profile but also the degree of extravasation and the clearance of extracellular QDs, making it an important variable for regulating the transfer and exchange process of QDs among three physiological compartments-bloodstream, extracellular space and Kupffer cells/hepatocytes.

Indigenous soil bacteria with the combined potential for hydrocarbon consumption and heavy metal resistance.

INTRODUCTION: Transconjugant bacteria with combined potential for hydrocarbon utilization and heavy metal resistance were suggested by earlier investigators for bioremediation of soils co-contaminated with hydrocarbons and heavy metals. The purpose of this study was to offer evidence that such microorganisms are already part of the indigenous soil microflora. METHODS: Microorganisms in pristine and oily soils were counted on nutrient agar and a mineral medium with oil as a sole carbon source, in the absence and presence of either sodium arsenate (As V), sodium arsenite (As III) or cadmium sulfate, and characterized via 16S rRNA gene sequencing. The hydrocarbon-consumption potential of individual strains in the presence and absence of heavy metal salts was measured. RESULTS: Pristine and oil-contaminated soil samples harbored indigenous bacteria with the combined potential for hydrocarbon utilization and As and Cd resistance in numbers up to 4 × 105 CFU g⁻¹. Unicellular bacteria were affiliated to the following species arranged in decreasing order of predominance: Bacillus subtilis, Corynebacterium pseudotuberculosis, Brevibacterium linens, Alcaligenes faecalis, Enterobacter aerogenes, and Chromobacterium orangum. Filamentous forms were affiliated to Nocardia corallina, Streptomyces flavovirens, Micromonospora chalcea, and Nocardia paraffinea. All these isolates could grow on a wide range of pure aliphatic and aromatic hydrocarbons, as sole sources of carbon and energy, and could consume oil and pure hydrocarbons in batch cultures. Low As concentrations, and to a lesser extent Cd concentrations, enhanced the hydrocarbon-consumption potential by the individual isolates. CONCLUSION: There is no need for molecularly designing microorganisms with the combined potential for hydrocarbon utilization and heavy metal resistance, because they are already a part of the indigenous soil microflora.

Abortiporus biennis tolerance to insoluble metal oxides: oxalate secretion, oxalate oxidase activity, and mycelial morphology.

The ability of Abortiporus biennis to tolerate and solubilize toxic metal oxides (Cu(2)O, Al(2)O(3), ZnO, CuFe(2)O(4)Zn, CdO, and MnO(2)) incorporated into agar media was investigated and the growth rate, oxalic acid secretion, and mycelial morphology were monitored. Among the tested metal oxides, formation of clear zones underneath the mycelium growing on Cu(2)O- and ZnO-amended plates was observed. ZnO, CdO and Cu(2)O caused the highest rate of fungal growth inhibition. An increased level of oxalic acid concentration was detected as a response of A. biennis to the presence of Cu(2)O, MnO(2), ZnO and CuFe(2)O(4)Zn in growth medium. The oxalate oxidase (OXO) was found to be responsible for oxalic acid degradation in A. biennis cultivated in metal-amended media. An increased level of OXO was observed in media amended with Cu(2)O, ZnO and MnO(2). Confocal microscopy used in this study revealed changes in mycelial morphology which appeared as increased hyphal branching, increased septation and increased spore number.

Effects of particulate and soluble cadmium species on biochemical and functional parameters in cultured murine macrophages.

Cultured murine macrophages (RAW 264.7) were used to evaluate the temporal relationships between cytotoxicity, phagocytosis, tumor necrosis factor-alpha (TNF-alpha), and nitric oxide (NO) production, and alterations in expression of stress proteins after exposure to cadmium oxide (CdO) or cadmium chloride (CdCl(2)), particulate and soluble forms of cadmium, respectively. Macrophages were exposed in vitro to CdO (25 or 50 microg) or CdCl(2) (30 or 40 microM) for 2 to 72 h. Cytotoxicity was not evident until 18 h when exposed to 30 microM CdCl(2) or 25 microg CdO, but occurred as early as 12 h after exposure to 40 microM CdCl(2) or 50 microg CdO. Relative to untreated controls, phagocytic activity decreased progressively from 2 to 24 h after exposure to both forms of cadmium. TNF-alpha levels increased to 2- to 3-fold after 4 h and remained elevated until 24 h after exposure to 25 and 50 microg CdO and 30 microM CdCl(2), but decreased by 18-24 h at 40 microM CdCl(2). CdCl(2) or CdO alone did not induce NO; however, both cadmium species reduced lipopolysaccharide (LPS)-stimulated NO production in a dose-dependent manner. Enhanced de novo synthesis of 70- and 90-kD heat shock, or stress, proteins was observed 2 to 8 h after exposure to both CdCl(2) and CdO; however, synthesis of these proteins returned to control levels by 24 h. Stress protein synthesis was enhanced by CdCl(2) or CdO prior to cytotoxicity, but coincided with a decrease in phagocytic capacity and an increase in TNF-a levels. The data suggest that cultured macrophages respond similarly in vitro to a particulate form and a soluble form of cadmium in a cell type that plays a pivotal role in inflammatory and immune responses.

Azuki bean cells are hypersensitive to cadmium and do not synthesize phytochelatins.

Suspension-cultured cells of azuki bean (Vigna angularis) as well as the original root tissues were hypersensitive to Cd (<10 microM). Repeated subculturings with a sublethal level of Cd (1-10 microM) did not affect the subsequent response of cells to inhibitory levels of Cd (10-100 microM). The azuki bean cells challenged to Cd did not contain phytochelatin (PC) peptides, unlike tomato (Lycopersicon esculentum) cells that have a substantial tolerance to Cd (>100 microM). Both of the cell suspensions contained a similar level of reduced glutathione (GSH) when grown in the absence of Cd. Externally applied GSH to azuki bean cells recovered neither Cd tolerance nor PC synthesis of the cells. Furthermore, enzyme assays in vitro revealed that the protein extracts of azuki bean cells had no activity converting GSH to PCs, unlike tomato. These results suggest that azuki bean cells are lacking in the PC synthase activity per se, hence being Cd hypersensitive. We concluded that the PC synthase has an important role in Cd tolerance of suspension-cultured cells.

Stabilization of adhesion plaques by the expression of drebrin A in fibroblasts.

The expression of drebrin A was induced in mouse fibroblasts (L cells) after transformation of cells with a vector that carried cDNA for rat drebrin A (developmentally regulated brain protein A) under the control of the promoter of the gene for metallothionein-I. When drebrin was expressed in the transformed cells (MTI-5 cells), the organization of actin filaments changed such that stress fibers were converted to a mesh-like structure. After subsequent treatment with 5 micrograms/ml cytochalasin D (a reagent that depolymerizes actin filaments), MTI-5 cells maintained their shape, while cells of a drebrin-negative cell line, MTI-11, formed retraction processes. Simultaneously, actin filaments changed into patchy dot-like aggregates in the cytoplasm of both MTI-5 and MTI-11 cells. These aggregates are known as cytoplasmic pools. In MTI-5 cells, adhesion plaques that were resistant to treatment with cytochalasin D appeared upon expression of drebrin. These adhesion plaques were immunostained with vinculin-specific antibodies, while those in MTI-11 cells were hardly immunostained. The amount of vinculin in MTI-5 cells increased in parallel with increase in the level of drebrin. These results suggest that expression of drebrin A induces changes in the assembly of actin filaments and adhesion plaques, with resultant modulation of cellular adhesion to the substratum.