Single gene silencing alters gene expression in both cycles and disrupts folate homeostasis. Folate over-supplementation and deficiency favors TS over MS cycle and causes prophase DNA damage.
Quercetin, a dietary flavonoid found in fruits and vegetables, has been described as a substance with many anti-cancer properties in a variety of preclinical investigations. In the present study, we demonstrate that 2D and 3D melanoma models exhibit not only different sensitivities to quercetin, but also opposite, cancer-promoting effects when metastatic melanoma spheroids are treated with quercetin. Higher concentrations of quercetin reduce melanoma growth in three tested cell lines, whereas low concentrations induce the opposite effect in metastatic melanoma spheroids but not in the non-metastatic cell line. High (>12.5 µM) or low (<6.3 µM) quercetin concentrations decrease or enhance cell viability, spheroid size, and cell proliferation, respectively. Additionally, melanoma cells cultivated in 2D already show significant caspase 3 activity at very low concentrations (>0.4 µM), whereas in 3D spheroids apoptotic cells, caspase 3 activity can only be detected in concentrations ≥12.5 µM. Further, we show that the tumor promoting or repressing effect in the 3D metastatic melanoma spheroids are likely to be elicited by a precisely controlled regulation of Nrf2/ARE-mediated cytoprotective genes, as well as ERK and NF-κB phosphorylation. According to the results obtained here, further studies are needed to better characterize the mechanisms of action underlying the pro- and anti-carcinogenic effects of quercetin on human melanomas.
Celiac disease (CD) is a chronic inflammatory condition caused by the ingestion of gliadin-containing food in genetically susceptible individuals. Undigested peptides of gliadin exert various effects, including increased intestinal permeability and inflammation in the small intestine. Although many therapeutic approaches are in development, a gluten-free diet is the only effective treatment for CD. Affecting at least 1% of the population in industrialized countries, it is important to generate therapeutic options against CD. Here, we describe the establishment of a high-throughput screening (HTS) platform based on AlphaLISA and electrical cell-substrate impedance sensing (ECIS) technology for the identification of anti-inflammatory and barrier-protective compounds in human enterocytes after pepsin-trypsin-digested gliadin (PT-gliadin) treatment. Our results show that the combination of these HTS technologies enables fast, reliable, simple, and label-free screening of IgY antibodies against PT-gliadin. Using this platform, we have identified a new chicken anti-PT-gliadin IgY antibody as a potential anti-CD agent.
Modern society is heavily dependent on critical raw materials, such as rare earth elements (REEs), for use in electronic devices. The increasing demand for these materials has led to the need for environmentally friendly methods of processing non-recycled materials from e-waste and wastewater, as well as waste streams from cleaning and manufacturing facilities. Modern society’s dependence on such materials is growing by the day, and with it, the need for environmentally friendly processing of non-recycled materials from e-waste and wastewater in the form of “end-of-life” products, as well as waste streams from cleaning and manufacturing facilities, also increases. As these are problematic indications for modern isolation methods in the industry, these sources may be more suitable for new techniques as they have low concentration and high throughput for bioaccumulation. Chemical methods using nanomaterials are already being tested for their possibilities but still depend on acids and harsh chemicals. Microorganisms, on the other hand, can adsorb/absorb REEs in a more ecological way. Previous studies could already show that it is possible to accumulate REEs in the precipitates of bacterial cultures spiked with REEs to a value of over 50%. However, the question arose whether rare earths were spun into the pellets by centrifugation, adsorbed, or really incorporated in the cells. Therefore, we established a new easy-to-use experimental design in which the microorganisms were spiked with an REE standard and washed to minimize the falsification of measurements by peripheral binding of ions before being analyzed for REE contents by ICP-OES. The bioaccumulation of rare earths in microorganisms was monitored, yielding an uptake rate of up to 53.12% of the overall present ionic REE concentration. In this manuscript, we present the different concentration measurements that were taken during the process, before and after washing of the cells, to create a full picture of the localization, binding, incorporation, and occurrence of the ions of interest. The setup also showed a correlation between the introduction method of rare earths and the uptake of certain elements that might be correlated with the differentiation between light and heavy rare earth elements, while Y and Sc often seem to represent outliers.
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