A multi-channel microfluidic platform based on human flavin-containing monooxygenase 3 for personalised medicine.

Human flavin-containing monooxygenase 3 (FMO3) is a drug-metabolizing enzyme (DME) which is known to be highly polymorphic. Some of its polymorphic variants are associated with inter-individual differences that contribute to drug response. In order to measure these differences, the implementation of a quick and efficient in vitro assay is highly desirable. To this end, in this work a microfluidic immobilized enzyme reactor (µ-IMER) was developed with four separate serpentines where FMO3 and its two common polymorphic variants (V257M and E158K) were covalently immobilized via glutaraldehyde cross-linking in the presence of a polylysine coating. Computational fluid dynamics simulations were performed to calculate the selected substrate retention time in serpentines with different surface areas at various flow rates. The oxidation of tamoxifen, an anti-breast cancer drug, was used as a model reaction to characterize the new device in terms of available surface area for immobilization, channel coating, and applied flow rate. The highest amount of product was obtained when applying a 10 µL min-1 flow rate on polylysine-coated serpentines with a surface area of 90 mm2 each. Moreover, these conditions were used to test the device as a multi-enzymatic platform by simultaneously assessing the conversion of tamoxifen by FMO3 and its two polymorphic variants immobilized on different serpentines of the same chip. The results obtained demonstrate that the differences observed in the conversion of tamoxifen within the chip are similar to those already published (E158K > WT > V257M). Therefore, this microfluidic platform provides a feasible option for fabricating devices for personalised medicine.

Occurrence, fate, and remediation for per-and polyfluoroalkyl substances (PFAS) in sewage sludge: A comprehensive review.

Addressing per-and polyfluoroalkyl substances (PFAS) contamination is an urgent environmental concern. While most research has focused on PFAS contamination in water matrices, comparatively little attention has been given to sludge, a significant by-product of wastewater treatment. This critical review presents the latest information on emission sources, global distribution, international regulations, analytical methods, and remediation technologies for PFAS in sludge and biosolids from wastewater treatment plants. PFAS concentrations in sludge matrices are typically in hundreds of ng/g dry weight (dw) in developed countries but are rarely reported in developing and least-developed countries due to the limited analytical capability. In comparison to water samples, efficient extraction and cleaning procedures are crucial for PFAS detection in sludge samples. While regulations on PFAS have mainly focused on soil due to biosolids reuse, only two countries have set limits on PFAS in sludge or biosolids with a maximum of 100 ng/g dw for major PFAS. Biological technologies using microbes and enzymes present in sludge are considered as having high potential for PFAS remediation, as they are eco-friendly, low-cost, and promising. By contrast, physical/chemical methods are either energy-intensive or linked to further challenges with PFAS contamination and disposal. The findings of this review deepen our comprehension of PFAS in sludge and have guided future research recommendations.

A review on microalgae-mediated biotechnology for removing pharmaceutical contaminants in aqueous environments: Occurrence, fate, and removal mechanism.

Pharmaceutical compounds in aquatic environments have been considered as emerging contaminants due to their potential risks to living organisms. Microalgae-based technology showed the feasibility of removing pharmaceutical contaminants. This review summarizes the occurrence, classification, possible emission sources, and environmental risk of frequently detected pharmaceutical compounds in aqueous environments. The efficiency, mechanisms, and influencing factors for the removal of pharmaceutical compounds through microalgae-based technology are further discussed. Pharmaceutical compounds frequently detected in aqueous environments include antibiotics, hormones, analgesic and non-steroidal anti-inflammatory drugs (NSAIDs), cardiovascular agents, central nervous system drugs (CNS), antipsychotics, and antidepressants, with a concentration ranging from ng/L to µg/L. Microalgae-based technology majorly remove the pharmaceutical compounds through bioadsorption, bioaccumulation, biodegradation, photodegradation, and co-metabolism. This review identifies the opportunities and challenges for microalgae-based technology and proposed suggestions for future studies to tackle challenges. The findings of this review advance our understanding of the occurrence and fate of pharmaceutical contaminants in aqueous environments, highlighting the potential of microalgae-based technology for pharmaceutical contaminants removal.

Ecotoxicological response of Spirulina platensis to coexisted copper and zinc in anaerobic digestion effluent.

Copper ion (Cu2+) and zinc ion (Zn2+) are widely co-existent in anaerobic digestion effluent as typical contaminants. This work aims to explore how Cu2+-Zn2+ association affects physiological properties of S. platensis using Schlösser medium (SM) and sterilized anaerobic digestion effluent (SADE). Microalgae cells viability, biochemical properties, uptake of Cu2+ and Zn2+, and risk assessment associated with the biomass reuse as additives to pigs were comprehensively assessed. Biomass production ranged from 0.03 to 0.28 g/L in SM and 0.63 to 0.79 g/L in SADE due to the presence of Cu2+ and Zn2+. Peak value of chlorophyll-a and carotenoid content during the experiment decreased by 70-100% and 40-100% in SM, and by 70-77% and 30-55% in SADE. Crude protein level reduced by 4-41% in SM and by 65-75% in SADE. The reduction ratio of these compounds was positively related to the Cu2+ and Zn2+ concentrations. Maximum value of saturated and unsaturated fatty acids was both obtained at 0.3 Cu + 2.0 Zn (50.8% and 22.8%, respectively) and 25% SADE reactors (33.8% and 27.7%, respectively). Uptake of Cu in biomass was facilitated by Zn2+ concentration (> 4.0 mg/L). Risk of S. platensis biomass associated with Cu2+ was higher than Zn2+. S. platensis from SM (Cu2+ ≤ 0.3 mg/L and Zn2+ ≤ 4.0 mg/L) and diluted SADE (25% and 50% SADE) reactors could be used as feed additives without any risk (hazard index <1), which provides sufficient protein and fatty acids for pig consumption. These results revealed the promising application of using S. platensis for bioremediation of Cu2+ and Zn2+ in anaerobic digestion effluent and harvesting biomass for animal feed additives.

Global identification of full-length cassava lncRNAs unveils the role of cold-responsive intergenic lncRNA 1 in cold stress response.

Long noncoding RNAs (lncRNAs) have been considered to be important regulators of gene expression in a range of biological processes in plants. A large number of lncRNAs have been identified in plants. However, most of their biological functions still remain to be determined. Here, we identified a total of 3004 lncRNAs in cassava under normal or cold-treated conditions from Iso-seq data. We further characterized a cold-responsive intergenic lncRNA 1 (CRIR1) as a novel positive regulator of the plant response to cold stress. CRIR1 can be significantly induced by cold treatment. Ectopic expression of CRIR1 in cassava enhanced the cold tolerance of transgenic plants. Transcriptome analysis demonstrated that CRIR1 regulated a range of cold stress-related genes in a CBF-independent pathway. We further found that CRIR1 RNA can interact with cassava cold shock protein 5 (MeCSP5), which acts as an RNA chaperone, indicating that CRIR1 may recruit MeCSP5 to improve the translation efficiency of messenger RNA. In summary, our study extends the repertoire of lncRNAs in plants as well as their role in cold stress responses. Moreover, it reveals a mechanism by which CRIR1 affected cold stress response by modulating the expression of stress-responsive genes and increasing their translational yield.

MeSPL9 attenuates drought resistance by regulating JA signaling and protectant metabolite contents in cassava.

KEY MESSAGE: Analysis of drought-related genes in cassava shows the involvement of MeSPL9 in drought stress tolerance and overexpression of a dominant-negative form of this gene demonstrates its negative roles in drought stress resistance. Drought stress severely impairs crop yield and is considered a primary threat to food security worldwide. Although the SQUAMOSA promoter binding protein-like 9 (SPL9) gene participates extensively in numerous developmental processes and in plant response to abiotic stimuli, its role and regulatory pathway in cassava (Manihot esculenta) response to the drought condition remain elusive. In the current study, we show that cassava SPL9 (MeSPL9) plays negative roles in drought stress resistance. MeSPL9 expression was strongly repressed by drought treatment. Overexpression of a dominant-negative form of miR156-resistant MeSPL9, rMeSPL9-SRDX, in which a 12-amino acid repressor sequence was fused to rMeSPL9 at the C terminus, conferred drought tolerance without penalizing overall growth. rMeSPL9-SRDX-overexpressing lines not only exhibited increased osmoprotectant metabolites including proline and anthocyanin, but also accumulated more endogenous jasmonic acid (JA) and soluble sugars. Transcriptomic and real-time PCR analysis suggested that differentially expressed genes were involved in sugar or JA biosynthesis, signaling, and metabolism in transgenic cassava under drought conditions. Exogenous application of JA further confirmed that JA conferred improved drought resistance and promoted stomatal closure in cassava leaves. Taken together, our findings suggest that MeSPL9 affects drought resistance by modulating protectant metabolite levels and JA signaling, which have substantial implications for engineering drought tolerant crops.