Uptake and Toxicity of Copper Oxide Nanoparticles in C6 Glioma Cells

Copper oxide nanoparticles (CuO-NPs) are frequently used for many technical applications, but are also known for their cell toxic potential. In order to investigate a potential use of CuO-NPs as a therapeutic drug for glioma treatment, we have investigated the consequences of an application of CuO-NPs on the cellular copper content and cell viability of C6 glioma cells. CuO-NPs were synthesized by a wet-chemical method and were coated with dimercaptosuccinic acid and bovine serum albumin to improve colloidal stability in physiological media. Application of these protein-coated nanoparticles (pCuO-NPs) to C6 cells caused a strong time-, concentration- and temperature-dependent copper accumulation and severe cell death. The observed loss in cellular MTT-reduction capacity, the loss in cellular LDH activity and the increase in the number of propidium iodide-positive cells correlated well with the specific cellular copper content. C6 glioma cells were less vulnerable to pCuO-NPs compared to primary astrocytes and toxicity of pCuO-NPs to C6 cells was only observed for incubation conditions that increased specific cellular copper contents above 20 nmol copper per mg protein. Both cellular copper accumulation as well as the pCuO-NP-induced toxicity in C6 cells were prevented by application of copper chelators, but not by endocytosis inhibitors, suggesting that liberation of copper ions from the pCuO-NPs is the first step leading to the observed toxicity of pCuO-NP-treated glioma cells.     

Biodistribution and Toxicity Studies of PRINT Hydrogel Nanoparticles in Mosquito Larvae and Cells

Mosquito-borne diseases continue to remain major threats to human and animal health and impediments to socioeconomic development. Increasing mosquito resistance to chemical insecticides is a great public health concern, and new strategies/technologies are necessary to develop the next-generation of vector control tools. We propose to develop a novel method for mosquito control that employs nanoparticles (NPs) as a platform for delivery of mosquitocidal dsRNA molecules to silence mosquito genes and cause vector lethality. Identifying optimal NP chemistry and morphology is imperative for efficient mosquitocide delivery. Toward this end, fluorescently labeled polyethylene glycol NPs of specific sizes, shapes (80 nm x 320 nm, 80 nm x 5000 nm, 200 nm x 200 nm, and 1000 nm x 1000 nm) and charges (negative and positive) were fabricated by Particle Replication in Non-Wetting Templates (PRINT) technology. Biodistribution, persistence, and toxicity of PRINT NPs were evaluated in vitro in mosquito cell culture and in vivo in Anopheles gambiae larvae following parenteral and oral challenge. Following parenteral challenge, the biodistribution of the positively and negatively charged NPs of each size and shape was similar; intense fluorescence was observed in thoracic and abdominal regions of the larval body. Positively charged NPs were more associated with the gastric caeca in the gastrointestinal tract. Negatively charged NPs persisted through metamorphosis and were observed in head, body and ovaries of adults. Following oral challenge, NPs were detected in the larval mid- and hindgut. Positively charged NPs were more efficiently internalized in vitro than negatively charged NPs. Positively charged NPs trafficked to the cytosol, but negatively charged NPs co-localized with lysosomes. Following in vitro and in vivo challenge, none of the NPs tested induced any cytotoxic effects.     

几种免疫效应细胞肿瘤杀伤途径的比较研究

比较了DCIK、CIK、NK与LAK在细胞来源、免疫表型、细胞因子分泌等生物学特性的差异,对效应细胞的肿瘤杀伤途径及其研究现状进行了概述和讨论.     

Stepwise Reconstitution of Interphase Microtubule Dynamics in Permeabilized Cells and Comparison to Dynamic Mechanisms in Intact Cells

Microtubules in permeabilized cells are devoid of dynamic activity and are insensitive to depolymerizing drugs such as nocodazole. Using this model system we have established conditions for stepwise reconstitution of microtubule dynamics in permeabilized interphase cells when supplemented with various cell extracts. When permeabilized cells are supplemented with mammalian cell extracts in the presence of protein phosphatase inhibitors, microtubules become sensitive to nocodazole. Depolymerization induced by nocodazole proceeds from microtubule plus ends, whereas microtubule minus ends remain inactive. Such nocodazole-sensitive microtubules do not exhibit subunit turnover. By contrast, when permeabilized cells are supplemented with Xenopus egg extracts, microtubules actively turn over. This involves continuous creation of free microtubule minus ends through microtubule fragmentation. Newly created minus ends apparently serve as sites of microtubule depolymerization, while net microtubule polymerization occurs at microtubule plus ends. We provide evidence that similar microtubule fragmentation and minus end–directed disassembly occur at the whole-cell level in intact cells. These data suggest that microtubule dynamics resembling dynamics observed in vivo can be reconstituted in permeabilized cells. This model system should provide means for in vitro assays to identify molecules important in regulating microtubule dynamics. Furthermore, our data support recent work suggesting that microtubule treadmilling is an important mechanism of microtubule turnover.     

Uptake of Intact Copper Oxide Nanoparticles Causes Acute Toxicity in Cultured Glial Cells

Copper oxide nanoparticles (CuO-NPs) dispersions are known for their high cell toxic potential but contaminating copper ions in such dispersions are a major hurdle in the investigation of specific nanoparticle-mediated toxicity. In order to distinguish between the adverse effects exhibited by CuO-NPs and/or by contaminating ionic copper, the membrane-impermeable copper chelator bathocuproine disulfonate (BCS) was added in a low molar ratio (20% of the total copper applied) in order to chelate the copper ions that had been released extracellularly from the CuO-NPs before or during the incubation. Physicochemical characterization of synthesized CuO-NPs revealed that the presence of this low concentration of BCS did not alter the size or zeta potential of the CuO-NPs. Application of CuO-NPs to C6 glioma cells and primary astrocytes induced a concentration- and temperature-dependent copper accumulation which was accompanied by a severe loss in cell viability. The adverse consequences of the CuO-NP application were not affected by the presence of 20% BCS, while the copper accumulation and cell toxicity observed after application of ionic copper were significantly lowered in the presence of BCS. These results demonstrate that for the experimental conditions applied the adverse consequences of an exposure of cultured glial cells to dispersions of CuO-NPs are mediated by accumulated NPs and not caused by the uptake of contaminating copper ions.

     

Ionic Mechanisms for Electrical Heterogeneity between Rabbit Purkinje Fiber and Ventricular Cells

The intrinsic heterogeneity of electrical action potential (AP) properties between Purkinje fibers (PFs) and the ventricular wall, as well as within the wall, plays an important role in ensuring successful excitation of the ventricles. It can also be proarrhythmic due to nonuniform repolarization across the Purkinje-ventricular junction. However, the ionic mechanisms that underlie the marked AP differences between PFs and ventricular cells are not fully characterized. We studied such mechanisms by developing a new family of biophysically detailed AP models for rabbit PF cells and three transmural ventricular cell types. The models were based on and validated against experimental data recorded from rabbit at ionic channel, single cell, and tissue levels. They were then used to determine the functional roles of each individual ionic channel current in modulating the AP heterogeneity at the rabbit Purkinje-ventricular junction, and to identify specific currents responsible for the differential response of PFs and ventricular cells to pharmacological interventions.     

In Vitro Toxicity Evaluation of 25-nm Anatase TiO2 Nanoparticles in Immortalized Keratinocyte Cells

Titanium dioxide (TiO(2)) nanoparticles (NPs) are massively fabricated and widely used in daily life, and thus potential risk has been posed to human health. However, the mechanism of the interaction between TiO(2) NPs and cells is still unclear. In this study, the interaction of anatase TiO(2) NPs with HaCaT cells is studied in vitro with multi-techniques. The TiO(2) NPs not only insert into cells through endocytic pathway but also penetrate into the cell. The TiO(2) NPs could produce reactive oxygen species (ROS) after dispersion spontaneously. Furthermore, the interaction between TiO(2) NPs and cellular components might also generate ROS. The ROS generation could lead to cellular toxicity if the level of ROS production overwhelms the antioxidant defense. Cytoskeletal components, particularly the microfilaments and microtubules, cause modifications upon exposure to TiO(2) NPs. With all results, the toxicological effects of TiO(2) NPs on HaCaT cell can be simplified into six events.     

Inbuilt Potential of YEM Medium and Its Constituents to Generate Ag/Ag2O Nanoparticles

We discovered that Yeast Extract Mannitol (YEM) medium possessed immense potential to generate silver nanoparticles from AgNO₃ upon autoclaving, which was evident from (i) alteration in color of the medium; (ii) peak at ∼410 nm in UV-Vis spectrum due to surface plasmon resonance specific to silver nanoparticles; and (iii) TEM investigations. TEM coupled with EDX confirmed that distinct nanoparticles were composed of silver. Yeast extract and mannitol were key components of YEM medium responsible for the formation of nanoparticles. PXRD analysis indicated crystalline geometry and Ag/Ag₂O phases in nanoparticles generated with YEM medium, yeast extract and mannitol. Our investigations also revealed that both mannitol and yeast extract possessed potential to convert ∼80% of silver ions in 0.5 mM AgNO₃ to nanoparticles, on autoclaving for 30 min at 121°C under a pressure of 1.06 kg/cm². Addition of filter sterilized AgNO₃ under ambient conditions to pre-autoclaved YEM medium and yeast extract brought about color change due to the formation of silver nanoparticles, but required prolonged duration. In general, even after 72 h intensity of color was significantly less than that recorded following autoclaving. Silver nanoparticles formed at room temperature were more heterogeneous compared to that obtained upon autoclaving. In summary, our findings demonstrated that (i) YEM medium and its constituents promote synthesis of silver nanoparticles; and (ii) autoclaving enhances rapid synthesis of silver nanoparticles by YEM medium, yeast extract and mannitol.     

Numerical Study of Plasmonic Effects of Ag Nanoparticles Embedded in the Active Layer on Performance Polymer Organic Solar Cells

In this paper, the light absorption in the active layer of polymer solar cells (OPV) by using plasmonic nanocrystals with a hexagonal lattice structure is investigated. To study the relationship between the performance of the OPV solar cell and its active layer, a three-dimensional model of its morphology is utilized. Therefore, the three-dimensional (3D) finite-difference time-domain method and Lumerical software were used to measure the field distribution and light absorption in the active layer in terms of wavelength. OPV solar cells with bilayer and bulk heterojunction structured cells were designed using hexagonal lattice crystals with plasmonic nanoparticles, as well as core–shell geometry to govern a design to optimize light trapping in the active layer. The parameters of shape, material, periodicity, size, and the thickness of the active layer as a function of wavelength in OPV solar cells have been investigated. A very thin active layer and an ultra-thin shell were used to achieve the highest increase in optical absorption. The strong alternating electromagnetic field around the core–shell plasmonic nanoparticles resulting from the localized surface plasmon resonance (LSPR) suggested by the Ag plasmonic nanocrystals increased the intrinsic optical absorption in the active layer poly(3-hexylthiophene):phenyl-C61-butyric acid methyl ester (P3HT:PCBM). Based on the photovoltaic results, the short circuit current ranged from 19.7 to 26.7 mA/cm².

     

High-Quantum Efficiency of the Plasmonic Photodetectors-Based on Coupling Between Ag Nanoparticles and Ag Nanograting Electrodes

The plasmonic effects due to the coupling of silver spherical nanoparticles with silver nanograting electrodes on active medium of the photo detector are investigated. Calculations show that the coupling of nanoparticles and nanograting electrodes at the surface of active medium lead to reflection wave into the active medium, and electrodes can participate in the absorption and quantum efficiency. Calculations show that the addition of nanograting electrodes to the structure can lead to increase the absorption coefficient up to 120% respect to the plasmonic structure containing only nanoparticles and about 4.5 times that of conventional (metal-semiconductor -metal ) MSM photo detectors.

     

Investigation on the Sensitivity and FOM of Ag Nanoparticles and Nanoarrays

The localized surface plasmon resonance (LSPR) spectroscopy of Ag nanoparticles (NPs) is sensitive to the changes of the surrounding medium, which enables the NPs to serve as plasmonic nanosensors. In this paper, the refractive index (RI) sensitivity and figure of merit (FOM) of individual NPs and nanoarrays are investigated by employing the finite difference time domain (FDTD) method. The influence of shape and size are analyzed for individual NPs, and the influence of particle spacing is analyzed for nanoarrays. It is found that the NP with shorter size in incident direction or longer size in polarization direction exhibits better sensing performance. And when the a_eff is between 20 and 60 nm, the larger NP exhibits higher sensitivity but lower FOM. The results of nanoarrays show that when particle spacing is large, the sensitivity of nanoarrays is large, and the sensitivity of nanoarrays decreases first and then increases as particle spacing decreases. In addition, the FOM of nanoarrays exhibits the similar trend.

     

Size-Dependent Toxicity of Silver Nanoparticles to Bacteria,Yeast, Algae,Crustaceans and Mammalian Cells In Vitro

The concept of nanotechnologies is based on size-dependent properties of particles in the 1–100 nm range. However, the relation between the particle size and biological effects is still unclear. The aim of the current paper was to generate and analyse a homogenous set of experimental toxicity data on Ag nanoparticles (Ag NPs) of similar coating (citrate) but of 5 different primary sizes (10, 20, 40, 60 and 80 nm) to different types of organisms/cells commonly used in toxicity assays: bacterial, yeast and algal cells, crustaceans and mammalian cells in vitro. When possible, the assays were conducted in ultrapure water to minimise the effect of medium components on silver speciation. The toxic effects of NPs to different organisms varied about two orders of magnitude, being the lowest (∼0.1 mg Ag/L) for crustaceans and algae and the highest (∼26 mg Ag/L) for mammalian cells. To quantify the role of Ag ions in the toxicity of Ag NPs, we normalized the EC₅₀ values to Ag ions that dissolved from the NPs. The analysis showed that the toxicity of 20–80 nm Ag NPs could fully be explained by released Ag ions whereas 10 nm Ag NPs proved more toxic than predicted. Using E. coli Ag-biosensor, we demonstrated that 10 nm Ag NPs were more bioavailable to E. coli than silver salt (AgNO₃). Thus, one may infer that 10 nm Ag NPs had more efficient cell-particle contact resulting in higher intracellular bioavailability of silver than in case of bigger NPs. Although the latter conclusion is initially based on one test organism, it may lead to an explanation for “size-dependent“ biological effects of silver NPs. This study, for the first time, investigated the size-dependent toxic effects of a well-characterized library of Ag NPs to several microbial species, protozoans, algae, crustaceans and mammalian cells in vitro.     

常用鱼药漂白粉和氯杀宁对背角无齿蚌的毒性影响

试验以洱海流域的背角无齿蚌为研究对象,确定常用鱼药:漂白粉、氯杀宁对湖泊大型底柄动物背角无齿蚌的毒性影响.试验得出漂白粉作用下,背角无齿蚌24 h、48 h、72 h、96 h、120 h、144 h、168 h的半致死率;氯杀宁作用下,背角无齿蚌24 h、48 h、72 h、96 h的半致死率;漂白粉有效氯安全浓度95.25～108.86 mg/L;氯杀宁有效氯安全浓度3.32～3.55 mg/L.通过显微镜观察,漂白粉和氯杀宁的主要作用部位为背角无齿蚌的呼吸系统,在其鳃上可见明显病变.     

Plasmonic and Energy Studies of Ag Nanoparticles in Silica-Titania Hosts

The present work reports an investigation of surface plasmon resonance (SPR) of silver nanoparticles in SiO₂–TiO₂ hosts. The surface plasmon resonance of silver nanoparticles was observed in the wavelength range 300–400 nm. Numerical calculation of SPR of silver nanoparticles with spherical morphology was done on the basis of discrete dipole approximation (DDA) method. The observed fluorescence spectrum fits well with the theoretically calculated one. The luminescence enhancement is attributed to the strong local electric field which increases the exciting and emitting photons coupled to SPR. An effort has been made to study the surface plasmon mediated excitation energy transfer (EET) between two spherical metal nanoparticles. The van der Waals (vdW) energy between plasmonic silver nanoparticles in the present hosts has been estimated.     

Polarization-Dependent Confocal Imaging of Individual Ag Nanorods and Nanoparticles

Polarization-dependent inelastic optical scattering (IOS) of individual Ag nanorods and nanoparticles are studied by confocal imaging. Stronger IOS is observed at two ends of the nanorod with laser polarizing parallel to the rod long axis while the IOS images of Ag nanoparticles elongate along laser polarization direction. The correlation between the far-field IOS image and near-field spatial distributions of the nanostructures′ electric field can be obtained. The IOS imaging is demonstrated to be an effective technique to study the optical properties of metal nanostructures, which also provides information for their applications in surface-enhanced Raman scattering.     

Cellular Mechanisms in Higher Plants Governing Tolerance to Cadmium Toxicity

Cadmium (Cd) is an inorganic mineral in the earth's crust. Cadmium entry into the environment occurs through geogenic and anthropogenic sources. Industrial activities including mining, electroplating, iron and steel plants, and battery production employ Cd during their processes and often release Cd into the environment. When disseminated into soil, Cd can be detrimental to agro-ecosystems because it is relatively mobile and phytotoxic even at low concentrations. Cadmium's phytotoxicity is due to reductions in the rate of transpiration and photosynthesis and chlorophyll concentration resulting in retardation of plant growth, and an alteration in the nutrient concentration in roots and leaves. In response to Cd toxicity, plants have developed protective cellular mechanisms such as synthesis of phytochelatins and metallothioneins, metal compartmentalization in vacuoles, and the increased activity of antioxidant enzymes to neutralize Cd-induced toxicity. While these direct protective mechanisms can help alleviate Cd toxicity, other indirect mechanisms such as microelements (zinc, iron, manganese, and selenium) interfering with Cd uptake may decrease Cd concentration in plants. This comprehensive review encompasses the significance of Cd, portals of contamination and toxicity to plants, and implications for crop production. Various mitigation strategies with the beneficial effects of zinc, iron, manganese, and selenium in activating defence mechanisms against Cd stress are discussed. Furthermore, this review systematically identifies and summarises suitable strategies for mitigating Cd-induced toxicity in plants.     

Apoptosis in Hemocytes Induces a Shift in Effector Mechanisms in the Drosophila Immune System and Leads to a Pro-Inflammatory State

Apart from their role in cellular immunity via phagocytosis and encapsulation, Drosophila hemocytes release soluble factors such as antimicrobial peptides, and cytokines to induce humoral responses. In addition, they participate in coagulation and wounding, and in development. To assess their role during infection with entomopathogenic nematodes, we depleted plasmatocytes and crystal cells, the two classes of hemocytes present in naïve larvae by expressing proapoptotic proteins in order to produce hemocyte-free (Hml-apo, originally called Hemo^less) larvae. Surprisingly, we found that Hml-apo larvae are still resistant to nematode infections. When further elucidating the immune status of Hml-apo larvae, we observe a shift in immune effector pathways including massive lamellocyte differentiation and induction of Toll- as well as repression of imd signaling. This leads to a pro-inflammatory state, characterized by the appearance of melanotic nodules in the hemolymph and to strong developmental defects including pupal lethality and leg defects in escapers. Further analysis suggests that most of the phenotypes we observe in Hml-apo larvae are alleviated by administration of antibiotics and by changing the food source indicating that they are mediated through the microbiota. Biochemical evidence identifies nitric oxide as a key phylogenetically conserved regulator in this process. Finally we show that the nitric oxide donor L-arginine similarly modifies the response against an early stage of tumor development in fly larvae.     

Bioenergetic Mechanisms in Astrocytes May Contribute to Amyloid Plaque Deposition and Toxicity

Alzheimer disease (AD) is characterized neuropathologically by synaptic disruption, neuronal loss, and deposition of amyloid β (Aβ) protein in brain structures that are critical for memory and cognition. There is increasing appreciation, however, that astrocytes, which are the major non-neuronal glial cells, may play an important role in AD pathogenesis. Unlike neurons, astrocytes are resistant to Aβ cytotoxicity, which may, in part, be related to their greater reliance on glycolytic metabolism. Here we show that, in cultures of human fetal astrocytes, pharmacological inhibition or molecular down-regulation of a main enzymatic regulator of glycolysis, 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase (PFKFB3), results in increased accumulation of Aβ within and around astrocytes and greater vulnerability of these cells to Aβ toxicity. We further investigated age-dependent changes in PFKFB3 and astrocytes in AD transgenic mice (TgCRND8) that overexpress human Aβ. Using a combination of Western blotting and immunohistochemistry, we identified an increase in glial fibrillary acidic protein expression in astrocytes that paralleled the escalation of the Aβ plaque burden in TgCRND8 mice in an age-dependent manner. Furthermore, PFKFB3 expression also demonstrated an increase in these mice, although at a later age (9 months) than GFAP and Aβ. Immunohistochemical staining showed significant reactive astrogliosis surrounding Aβ plaques with increased PFKFB3 activity in 12-month-old TgCRND8 mice, an age when AD pathology and behavioral deficits are fully manifested. These studies shed light on the unique bioenergetic mechanisms within astrocytes that may contribute to the development of AD pathology.     

Toxicity of CdSe Nanoparticles in Caco-2 Cell Cultures

Background  

Potential routes of nanomaterial exposure include inhalation, dermal contact, and ingestion. Toxicology of inhalation of ultra-fine particles has been extensively studied; however, risks of nanomaterial exposure via ingestion are currently almost unknown. Using enterocyte-like Caco-2 cells as a small intestine epithelial model, the possible toxicity of CdSe quantum dot (QD) exposure via ingestion was investigated. Effect of simulated gastric fluid treatment on CdSe QD cytotoxicity was also studied.