ABSTRACT
Single-cell inductively coupled plasma mass spectrometry (SC-ICP-MS) has become a powerful and fast tool to evaluate the elemental composition at a single-cell level. In this study, the cellular bioavailability of arsenite (incubation of 25 and 50 µM for 0-48 h) has been successfully assessed by SC-ICP-MS/MS for the first time directly after re-suspending the cells in water. This procedure avoids the normally arising cell membrane permeabilization caused by cell fixation methods (e.g. methanol fixation). The reliability and feasibility of this SC-ICP-MS/MS approach with a limit of detection of 0.35 fg per cell was validated by conventional bulk ICP-MS/MS analysis after cell digestion and parallel measurement of sulfur and phosphorus.
Subject(s)
Arsenites/metabolism , Single-Cell Analysis/methods , Tandem Mass Spectrometry/methods , A549 Cells , Arsenites/analysis , Biological Availability , Humans , Phosphorus/analysis , Sulfur/analysisABSTRACT
Arsenic-containing hydrocarbons (AsHCs), natural products found in seafood, have recently been shown to exert toxic effects in human neurons. In this study we assessed the toxicity of three AsHCs in cultured human astrocytes. Due to the high cellular accessibility and substantial toxicity observed astrocytes were identified as further potential brain target cells for arsenolipids. Thereby, the AsHCs exerted a 5-19-fold higher cytotoxicity in astrocytes as compared to arsenite. Next we compared the toxicity of the arsenicals in a co-culture model of the respective human astrocytes and neurons. Notably the AsHCs did not show any substantial toxic effects in the co-culture, while arsenite did. The arsenic accessibility studies indicated that in the co-culture astrocytes protect neurons against cellular arsenic accumulation especially after incubation with arsenolipids. In summary, these data underline the importance of the glial-neuron interaction when assessing the in vitro neurotoxicity of new unclassified metal species.
Subject(s)
Arsenic Poisoning/etiology , Arsenicals/chemistry , Astrocytes/drug effects , Lipids/chemistry , Lipids/toxicity , Arsenic Poisoning/metabolism , Arsenicals/metabolism , Astrocytes/cytology , Astrocytes/metabolism , Cell Culture Techniques , Cell Line , Cell Survival/drug effects , Coculture Techniques , Humans , Lipid Metabolism , Neurons/cytology , Neurons/drug effects , Neurons/metabolismSubject(s)
Bacterial Vaccines , Cattle Diseases/prevention & control , Salmonella Infections, Animal/prevention & control , Salmonella typhimurium/immunology , Administration, Oral , Animals , Antibodies, Bacterial/biosynthesis , Bacterial Vaccines/administration & dosage , Cattle , Female , Immunization/veterinary , Male , Vaccines, Attenuated/administration & dosageABSTRACT
The two-dimensional immunoelectrophoresis was used in an attempt to differentiate between the species of the genus Bacillus and reveal their taxonomic relationship. Three strains each of the species B. subtilis, B. licheniformis, B. cereus and one strain each of the species B. laterosporus and B. sphaericus were selected. The substrate investigated was ultrasonic extract. The evaluation of the immunoelectropherogrames was based on the numbers of the precipitates as well as on the arrangement and shape of the precipitation lines. In most cases it was possible to identify the strains belonging to a certain species because of the fact, that in the homologeous system considerable more precipitates occurred than in heterologeous systems and in addition the optical pattern of the precipitation showed up with species-specific characteristics. The extent of the divergence between the homologic and a given heterologic pattern mirrored frequently the degree of affinity of the strain involved.
