Conventional wastewater treatment with primary and secondary treatment processes efficiently remove microplastics (MPs) from the wastewater. Despite the efficient removal, final effluents can act as entrance route of MPs, given the large volumes constantly discharged into the aquatic environments. This study investigated the removal of MPs from effluent in four different municipal wastewater treatment plants utilizing different advanced final-stage treatment technologies. The study included membrane bioreactor treating primary effluent and different tertiary treatment technologies (discfilter, rapid sand filtration and dissolved air flotation) treating secondary effluent. The MBR removed 99.9% of MPs during the treatment (from 6.9 to 0.005 MP L), rapid sand filter 97% (from 0.7 to 0.02 MP L), dissolved air flotation 95% (from 2.0 to 0.1 MP L) and discfilter 40-98.5% (from 0.5 - 2.0 to 0.03-0.3 MP L) of the MPs during the treatment. Our study shows that with advanced final-stage wastewater treatment technologies WWTPs can substantially reduce the MP pollution discharged from wastewater treatment plants into the aquatic environments.

Wastewater treatment plants (WWTPs) can offer a solution to reduce the point source input of microlitter and microplastics into the environment. To evaluate the contributing processes for microlitter removal, the removal of microlitter from wastewater during different treatment steps of mechanical, chemical and biological treatment (activated sludge) and biologically active filter (BAF) in a large (population equivalent 800 000) advanced WWTP was examined. Most of the microlitter was removed already during the pre-treatment and activated sludge treatment further decreased the microlitter concentration. The overall retention capacity of studied WWTP was over 99% and was achieved after secondary treatment. However, despite of the high removal performance, even an advanced WWTP may constitute a considerable source of microlitter and microplastics into the aquatic environment given the large volumes of effluent discharged constantly. The microlitter content of excess sludge, dried sludge and reject water were also examined. According to the balance analyses, approximately 20% of the microlitter removed from the process is recycled back with the reject water, whereas 80% of the microlitter is contained in the dried sludge. The study also looked at easy microlitter sampling protocol with automated composite samplers for possible future monitoring purposes.

Age-related macular degeneration (AMD) is a multi-factorial disease that is the leading cause of irreversible and severe vision loss in the developed countries. It has been suggested that the pathogenesis of dry AMD involves impaired protein degradation in retinal pigment epithelial cells (RPE). RPE cells are constantly exposed to oxidative stress that may lead to the accumulation of damaged cellular proteins, DNA and lipids and evoke tissue deterioration during the aging process. The ubiquitin-proteasome pathway and the lysosomal/autophagosomal pathway are the two major proteolytic systems in eukaryotic cells. NRF-2 (nuclear factor-erythroid 2-related factor-2) and PGC-1α (peroxisome proliferator-activated receptor gamma coactivator-1 alpha) are master transcription factors in the regulation of cellular detoxification. We investigated the role of NRF-2 and PGC-1α in the regulation of RPE cell structure and function by using global double knockout (dKO) mice. The NRF-2/PGC-1α dKO mice exhibited significant age-dependent RPE degeneration, accumulation of the oxidative stress marker, 4-HNE (4-hydroxynonenal), the endoplasmic reticulum stress markers GRP78 (glucose-regulated protein 78) and ATF4 (activating transcription factor 4), and damaged mitochondria. Moreover, levels of protein ubiquitination and autophagy markers p62/SQSTM1 (sequestosome 1), Beclin-1 and LC3B (microtubule associated protein 1 light chain 3 beta) were significantly increased together with the Iba-1 (ionized calcium binding adaptor molecule 1) mononuclear phagocyte marker and an enlargement of RPE size. These histopathological changes of RPE were accompanied by photoreceptor dysmorphology and vision loss as revealed by electroretinography. Consequently, these novel findings suggest that the NRF-2/PGC-1α dKO mouse is a valuable model for investigating the role of proteasomal and autophagy clearance in the RPE and in the development of dry AMD.

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The microRNA (miRNA) cargo contained in plasma extracellular vesicles (EVs) offers a relatively little explored source of biomarkers for brain diseases that can be obtained noninvasively. Methods to isolate EVs from plasma, however, are still being developed. For EV isolation, it is important to ensure the removal of vesicle-free miRNAs, which account for approximately two-thirds of plasma miRNAs. Membrane particle precipitation-based EV isolation is an appealing method because of the simple protocol and high yield. Here, we evaluated the performance of a precipitation-based method to obtain enriched EV-specific miRNAs from a small volume of rat plasma. We performed size-exclusion chromatography (SEC) on precipitation-isolated EV pellets and whole plasma. The SEC fractions were analysed using Nanoparticle Tracking Analysis (NTA), protein and miRNA concentration assays, and droplet digital polymerase chain reaction for four miRNAs (miR-142-3p, miR-124-3p, miR-23a, miR-122). Precipitation-isolated EVs and selected SEC fractions from the plasma were also analysed with transmission electron microscopy (TEM). Precipitation-based EV isolation co-precipitated 9% to 15% of plasma proteins and 21% to 99% of vesicle-free miRNAs, depending on the individual miRNAs. In addition, the amount of miR-142-3p, found mainly in EV fractions, was decreased in the EV fractions, indicating that part of it was lost during precipitation-based isolation. Western blot and TEM revealed both protein and lipoprotein contamination in the precipitation-isolated EV-pellets. Our findings indicate that a precipitation-based method is not sufficient for purifying plasma EV-contained miRNA cargo. The particle number measured by NTA is high, but this is mostly due to the contaminating lipoproteins. Although a part of the vesicle-free miRNA is removed, vesicle-free miRNA still dominates in plasma EV pellets isolated by the precipitation-based method.