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There is a growing concern nowadays over the exposure of nanomaterials and their effects in aquatic life. In spite of reporting the changes in physiology, reproduction and behaviour in fish by different nanoparticles, the molecular events underlying in the aquatic bodies due to the toxicity of zinc oxide nanoparticles (ZnO NPs) are mainly unexplored. Therefore, the present study carried out an ex vivo exposure of ZnO NPs at various concentrations (0.382, 0.573 and 1.146 mg L-1) in freshwater fish Cyprinus carpio to investigate the potential adverse effects. The results revealed that ZnO NPs exposure altered the haematological parameter and induces the reactive oxygen species (ROS) that leads to elevation of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidise (GPx), glutathione S-transferase (GST) and reduced glutathione (GSH) activity in C. carpio. Furthermore, histopathological analysis exhibited that the ZnO NPs caused lamellar fusion, aneurism, cytoplasmic vacuolation, nuclear alteration, necrotic muscle fiber and pyknotic nuclei in the gills, liver and muscles of C. carpio. ZnO NPs exposure significantly up-regulated the overlapping expressions of SOD1, CAT, GPx1a, GST-α, CYP1A, and Nrf-2 genes. A higher level of Zn bioaccumulation was observed in the following order: gill (35.03 ± 2.50 µg g-1), liver (5.33 ± 0.73 µg g-1) and muscle (2.30 ± 0.20 µg g-1) at 1.146 mg L-1 exposure of ZnO NPs. Hence, the current study indicated that the biogenic ZnO NPs generate toxicity in fishes by modifying the antioxidant defense mechanisms, histomorphology, and oxidative stress encoding genes.

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We are standardizing protocols to develop egg white (EW) as a cost-effective platform for culture of three-dimensional (3-D) multicellular tumor spheroids for application in understanding tumor microenvironments and drug screening. In this article, we describe several physical and physiological characteristics of EW to use it as 3-D cell culture platform. Field emission scanning electron microscopy revealed the presence of different microstructures. Hydrodynamic size distribution data indicated nano- and micron-sized particles. Rheological measurements revealed the viscosity and viscoelastic behavior appropriate for maintaining cell viability and supporting 3-D cell growth under high-sheer conditions. It was found that thereis no autofluorescence, a requirement for imparting transparency and for microscopic observations of the spheroids. The EW facilitated the development of 3-D tumor spheroids, with an emphasis of difference in cell proliferation and intercellular cytoskeletal organization between two-dimensional and 3-D spheroid cultures. Put together, EW proves to be a cost-affordable and simple platform for 3-D culture of tumor spheroids.

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The mechanisms underlying cobalt toxicity in aquatic species in general and cnidarians in particular remain poorly understood. Herein we investigated cobalt toxicity in a Hydra model from morphological, histological, developmental, and molecular biological perspectives. Hydra, exposed to cobalt (0-60 mg/L), were altered in morphology, histology, and regeneration. Exposure to standardized sublethal doses of cobalt impaired feeding by affecting nematocytes, which in turn affected reproduction. At the cellular level, excessive ROS generation, as the principal mechanism of action, primarily occurred in the lysosomes, which was accompanied by the upregulation of expression of the antioxidant genes SOD, GST, GPx, and G6PD. The number of Hsp70 and FoxO transcripts also increased. Interestingly, the upregulations were higher in the 24-h than in the 48-h time-point group, indicating that ROS overwhelmed the cellular defense mechanisms at the latter time-point. Comet assay revealed DNA damage. Cell cycle analysis indicated the induction of apoptosis accompanied or not by cell cycle arrest. Immunoblot analyses revealed that cobalt treatment triggered mitochondria-mediated apoptosis as inferred from the modulation of the key proteins Bax, Bcl-2, and caspase-3. From this data, we suggest the use of Hydra as a model organism for the risk assessment of heavy metal pollution in aquatic ecosystems.

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Nanotechnology has emerged as a powerful field of applied research. However, the potential toxicity of nano-materials is a cause of concern. A thorough toxicological investigation is required before a nanomaterial is evaluated for application of any kind. In this context, there is concerted effort to find appropriate test systems to assess the toxicity of nanomaterials. Toxicity of a nanomaterial greatly depends on its physicochemical properties and the biological system with which it interacts. The present research was carried out with a view to generate data on eco-toxicological impacts of copper oxide nanorod (CuO NR) in Hydra magnipapillata 105 at organismal, cellular and molecular levels. Exposure of hydra to CuO NR resulted in severe morphological alterations in a concentration- as well as duration-dependent manner. Impairment of feeding, population growth, and regeneration was also observed. In vivo and in vitro analyses revealed induction of oxidative stress, genotoxicity, and molecular machinery of apoptotic cell death, accompanied by disruption of cell cycle progression. Taken together, CuO nanorod is potentially toxic to the biological systems. Also, hydra offers potential to be used as a convenient model organism for aquatic ecotoxicological risk assessment of nanomaterials.

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Copper, an essential microelement, is known to be toxic to aquatic life at concentrations higher than that could be tolerated. Copper-induced oxidative stress has been documented in vitro, yet the in vivo effects of metal-induced oxidative stress have not been extensively studied in the lower invertebrates. The objective of the present study has been to find the effect of ROS-mediated toxicity of environmentally relevant concentrations of copper at organismal and cellular levels in Hydra magnipapillata. Exposure to copper at sublethal concentrations (0.06 and 0.1mg/L) for 24 or 48h resulted in generation of significant levels of intracellular reactive oxygen species (ROS). We infer that the free radicals here originate predominantly at the lysosomes but partly at the mitochondria also as visualized by H2-DHCFDA staining. Quantitative real-time PCR of RNA extracted from copper-exposed polyps revealed dose-dependent up-regulation of all antioxidant response genes (CAT, SOD, GPx, GST, GR, G6PD). Concurrent increase of Hsp70 and FoxO genes suggests the ability of polyps to respond to stress, which at 48h was not the same as at 24h. Interestingly, the transcript levels of all genes were down-regulated at 48h as compared to 24h incubation period. Comet assay indicated copper as a powerful genotoxicant, and the DNA damage was dose- as well as duration-dependent. Western blotting of proteins (Bax, Bcl-2 and caspase-3) confirmed ROS-mediated mitochondrial cell death in copper-exposed animals. These changes correlated well with changes in morphology, regeneration and aspects of reproduction. Taken together, the results indicate increased production of intracellular ROS in Hydra on copper exposure.

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