Systemically Achievable Doses of Beer Flavonoids Induce Estrogenicity in Human Endometrial Cells and Cause Synergistic Effects With Selected Pesticides

Some prenylated polyphenols originating from hops, which are thus natural constituents of beer, have been discussed critically for their agonistic potential toward estrogen receptors. So far, little attention has been attributed to the fact that humans are typically not exposed to isolated compounds, but to mixtures which for example might comprise in addition to hop flavonoids further xenoestrogens, e.g., certain pesticides used for plant protection of hops and barley. Thus, we used the alkaline phosphatase assay to assess combinatory estrogenic effects of three signature compounds – xanthohumol, 8-prenylnaringenin and iso-xanthohumol–on Ishikawa cells in a combination that resembled the concentration ratios observable in beer. Moreover, we added this natural flavonoid pattern to a mixture of representative estrogenic pesticides to assess their combined effects. Using state-of-the-art statistical tools, we observed cumulative to slightly synergistic effects between isolated flavonoids as well as the flavonoid and the pesticide mixture. Of potential importance, these effects were found at low nanomolar hop polyphenol concentrations that one can reasonably expect to occur in vivo after the consumption of strongly hopped beer. Taken together, our results imply that cumulative/synergistic estrogenicity should be explored in detail and urgently be incorporated into risk assessment of prenylated chalcones.

An Exonuclease I-Aided Turn-Off Fluorescent Strategy for Alkaline Phosphatase Assay Based on Terminal Protection and Copper Nanoparticles

As an important DNA 3′-phosphatase, alkaline phosphatase can repair damaged DNA caused by replication and recombination. It is essential to measure the level of alkaline phosphatase to indicate some potential diseases, such as cancer, related to alkaline phosphatase. Here, we designed a simple and fast method to detect alkaline phosphatase quantitively. When alkaline phosphatase is present, the resulting poly T-DNA with a 3′-hydroxyl end was cleaved by exonuclease I, prohibiting the formation of fluorescent copper nanoparticles. However, the fluorescent copper nanoparticles can be monitored with the absence of alkaline phosphatase. Hence, we can detect alkaline phosphatase with this turn-off strategy. The proposed method is able to quantify the concentration of alkaline phosphatase with the LOD of 0.0098 U/L. Furthermore, we utilized this method to measure the effects of inhibitor Na3VO4 on alkaline phosphatase. In addition, it was successfully applied to quantify the level of alkaline phosphatase in human serum. The proposed strategy is sensitive, selective, cost effective, and timesaving, having a great potential to detect alkaline phosphatase quantitatively in clinical diagnosis.

Mesoporous Bioactive Glasses Cytocompatibility Assessment: A Review of In Vitro Studies

Thanks to their high porosity and surface area, mesoporous bioactive glasses (MBGs) have gained significant interest in the field of medical applications, in particular, with regards to enhanced bioactive properties which facilitate bone regeneration. The aim of this article is to review the state of the art regarding the biocompatibility evaluation of MBGs and provide a discussion of the various approaches taken. The research was performed using PubMed database and covered articles published in the last five years. From a total of 91 articles, 63 were selected after analyzing them according to our inclusion and exclusion criteria. In vitro methodologies and techniques used for biocompatibility assessment were investigated. Among the biocompatibility assessment techniques, scanning electron microscopy (SEM) has been widely used to study cell morphology and adhesion. Viability and proliferation were assessed using different assays including cell counting and/or cell metabolic activity measurement. Finally, cell differentiation tests relied on the alkaline phosphatase assay; however, these were often complemented by specific bimolecular tests according to the exact application of the mesoporous bioactive glass. The standardization and validation of all tests performed for MBG cytocompatibility is a key aspect and crucial point and should be considered in order to avoid inconsistencies, bias between studies, and unnecessary consumption of time. Therefore, introducing standard tests would serve an important role in the future assessment and development of MBG materials.

A two fluorescent signal indicator-based ratio fluorometric alkaline phosphatase assay based on one signal precursor

A simple ratio fluorescent ALP biosensor based on MnO2 nanosheet-triggered in situ generation of two fluorescent signal indicators.

Target-induced transcription of light-up RNA aptamer to construct a novel method for alkaline phosphatase assay

We herein developed a novel method for alkaline phosphatase (ALP) assay based on the target-induced transcription of light-up RNA aptamer, consequently producing highly enhanced fluorescence signal upon specifically binding to...

Antibodies recognizing the C terminus of PP2A catalytic subunit are unsuitable for evaluating PP2A activity and holoenzyme composition

The methyl-esterification of the C-terminal leucine of the protein phosphatase 2A (PP2A) catalytic (C) subunit is essential for the assembly of specific trimeric PP2A holoenzymes, and this region of the C subunit also contains two threonine and tyrosine phosphorylation sites. Most commercial antibodies—including the monoclonal antibody 1D6 that is part of a frequently used, commercial phosphatase assay kit—are directed toward the C terminus of the C subunit, raising questions as to their ability to recognize methylated and phosphorylated forms of the enzyme. Here, we tested several PP2A C antibodies, including monoclonal antibodies 1D6, 7A6, G-4, and 52F8 and the polyclonal antibody 2038 for their ability to specifically detect PP2A in its various modified forms, as well as to coprecipitate regulatory subunits. The tested antibodies preferentially recognized the nonmethylated form of the enzyme, and they did not coimmunoprecipitate trimeric holoenzymes containing the regulatory subunits B or B′, an issue that precludes their use to monitor PP2A holoenzyme activity. Furthermore, some of the antibodies also recognized the phosphatase PP4, demonstrating a lack of specificity for PP2A. Together, these findings suggest that reinterpretation of the data generated by using these reagents is required.

Anodic electrochemiluminescence from CsPbBr3 perovskite quantum dots for an alkaline phosphatase assay

Fundamental mechanisms of ECL generation in a CsPbBr3 QDs/H2A system.