Long-Term Toxicological Monitoring of a Multibarrier Advanced Wastewater Treatment Plant Comprising Ozonation and Granular Activated Carbon with In Vitro Bioassays

A set of CALUX in vitro bioassays was applied for long-term toxicity monitoring at an advanced wastewater treatment plant comprising ozonation and granular activated carbon filtration for the abatement of contaminants of emerging concern (CEC). During the 13-month monitoring, eight reporter gene assays targeting different modes of action along the cellular toxicity pathway were accessed to evaluate the suitability and robustness of the technologies. Two approaches were followed: on the one hand, signal reduction during advanced treatment was monitored; on the other hand, results were compared to currently available effect-based trigger values (EBTs). A decrease of the corresponding biological equivalent concentrations after the multibarrier system could be observed for all modes of action; while the estrogenic activity decreased below the EBT already during ozonation, the potencies of oxidative stress-like and toxic PAH-like compounds still exceeded the discussed EBT after advanced treatment. Overall, the long-term monitoring confirmed the positive effect of the multibarrier system, commonly evaluated only by CEC abatement based on chemical analysis. It could be demonstrated that advanced WWTPs designed for CEC abatement are suitable to significantly decrease toxicity responses not only in the frame of pilot studies but under real-world conditions as well.

Gui Shao Di Huang Wan promotes angiogenesis and regulates oestrogen receptor α and progesterone receptor β in in vitro bioassays

Background and aim: The formula Gui Shao Di Huang Wan (GSDW) is used frequently to treat female infertility. This study aims to investigate some of the possible mechanisms of action of GSDW using in vitro bioassays of angiogenesis in Human Uterine Microvascular Endothelial Cells (HUVEC) and Human Uterine Microvascular Endothelial Cells (HUtMEC) and ovarian steroid receptor expression in a human endometrial cell line (Ishikawa). Experimental procedure: Aqueous extracts of GSDW and its component herbs were tested for pro-angiogenic activity using both HUVEC and HUtMEC 2D differentiation assays performed on Matrigel and effects on HUVEC proliferation using the MTT assay. Effects on the expression of Estrogen Receptor alpha (ER alpha) and Progesterone Receptor beta (PRbeta) in Ishikawa cells were determined using immunoblotting. Results and Conclusion: ll analysed parameters of differentiation were increased by GSDW in both the HUVEC and HUtMEC mesh. Furthermore, measures of total length, segment number, junction number were affected by some but not all component herbs. The MTT assay showed an increase in proliferation of HUVECs at concentrations of GSDW between 0.68 and 5.47 ug/mL at 48 and 72 hours. In Ishikawa cells downregulation of ER alpha; and upregulation of PR beta; was seen after 48 hours incubation with 4 of the 8 herbs in the formula. The findings in this study demonstrate that GSDW has the potential to affect key parameters (vascular, sex hormone receptor expression) in vitro. This offers a mechanism by which these herbs may enhance fertility through improved endometrial receptivity and pregnancy rates.