Microplastics in human urine: Characterisation using µFTIR and sampling challenges using healthy donors and endometriosis participants.

Microplastics (MPs) are found in all environments, within the human food chain, and have been recently detected in several human tissues. The objective herein was to undertake an analysis of MP contamination in human urine samples, from healthy individuals and participants with endometriosis, with respect to their presence, levels, and the characteristics of any particles identified. A total of 38 human urine samples and 15 procedural blanks were analysed. MPs were characterised using µFTIR spectroscopy (size limitation of 5 µm) and SEM-EDX. In total, 123 MP particles consisting of 22 MP polymer types were identified within 17/29 of the healthy donor (10 mL) urine samples, compared with 232 MP particles of differing 16 MP polymer types in 12/19 urine samples from participants with endometriosis. Healthy donors presented an unadjusted average of 2589 ± 2931 MP/L and participants with endometriosis presented 4724 ± 9710 MP/L. Polyethylene (PE)(27%), polystyrene (PS)(16%), resin and polypropylene (PP)(both 12%) polymer types were most abundant in healthy donor samples, compared with polytetrafluoroethylene (PTFE) (59%), and PE (16%) in samples from endometriosis participants. The MP levels within healthy and endometriosis participant samples were not significantly different. However, the predominant polymer types varied, and the MPs from the metal catheter-derived endometriosis participant samples and healthy donors were significantly smaller than those observed in the procedural blanks. The procedural blank samples comprised 62 MP particles of 10 MP polymer types, mainly PP (27%), PE (21%), and PS (15%) with a mean ± SD of 17 ± 18, highlighting the unavoidable contamination inherent in measurement of MPs from donors. This is the first evidence of MP contamination in human urine with polymer characterisation and accounting for procedural blanks. These results support the phenomenon of transport of MPs within humans, specifically to the bladder, and their characterisation of types, shapes and size ranges identified therein.

Cytotoxic effects of Ridolfia segetum (L.) Moris phytoproducts in cancer cells.

ETHNOPHARMACOLOGICAL RELEVANCE: The past few years have witnessed an increasing interest in essential oils (EOs) as potential therapeutic agents against a wide variety of pathologies, including cancer. EOs extracted from Ridolfia segetum (L.) Moris (R. segetum) are a clear example of a phytoproduct with therapeutic applications, as it is widely used in traditional medicine due to its antioxidant and anti-inflammatory properties, and these properties were already validated by previous studies. Although, it is well established that inflammation is a key hallmark of cancer, with a key role promoting tumorigenesis, and being chronic inflammation often associated with tumorigenic processes, there are no previous studies regarding the assessment of the antitumoural potential of R. segetum EOs. AIM OF THE STUDY: The present study intends to be the first to evaluate the antitumoural proprieties of R. segetum EO phytoproducts in cancer cell models. MATERIALS AND METHODS: For this, R. segetum EOs were extracted from plants collected at either flowering (RS_Fl) or fruiting (RS_Fr) stage. The impact on proliferation and viability of treatment with R. segetum EO extracts was assessed using in vitro 2D and 3D models. RESULTS: Both R. segetum EOs presented effective antiproliferative/viability effects, evidence noted by low IC50 values in 2D models, and significant reduction of spheroid size in 3D in vitro models. Mechanistically, treatment with R. segetum EOs was associated with an altered G1 (associated with p21 stabilisation), and subsequent induction of apoptosis. CONCLUSIONS: Overall, these results indicate that R. segetum EOs have potential as suitable antitumoural therapeutic agents.

Secondary metabolites (essential oils) from sand-dune plants induce cytotoxic effects in cancer cells.

ETHNOPHARMACOLOGICAL RELEVANCE: Despite advances in modern therapeutic strategies, cancer remains the second leading cause of death worldwide. Therefore, there is a constant need to develop more efficient anticancer targeting strategies. The anticancer therapeutic proprieties of medicinal plants and their bioactive compounds have been reported for several years, making natural extracts and/or compounds derived from these a promising source of novel anticancer agents. Sand dune plants are subjected to severe environmental stresses, leading to the development of adaptations, including the production of secondary metabolites with a wide range of bioactivities, such as: anti-inflammatory, analgesic, antiseptic, hypoglycaemic, hypotensive, antinociceptive, antioxidant and anticancer. AIM OF THE STUDY: The anticancer potential of sand dune plants remains under-investigated, so this research describes the characterisation of the composition of bioactive EOs from sand-dune plants of Peniche (Portugal), and assessment of their activity in vitro and potential mechanism of action. MATERIALS AND METHODS: EOs were extracted from six sand-dune species of plants from Peniche sand dunes: Crithmum maritimum L., Seseli tortuosum L., Artemisia campestris subsp. maritima (DC.) Arcang., Juniperus phoenicea var. turbinata (Guss.) Parl., Otanthus maritimus (L.) Hoffmanns. & Link, and Eryngium maritimum L.. EOs composition was fully characterised chemically using Gas Chromatography-Mass Spectrometry (GC-MS). The assessment of anticancer activity and mechanism of action was performed in vitro using breast and colorectal cancer 2D and 3D spheroid cell line models, through cell proliferation assay, western blotting analysis, and cell cycle analysis. RESULTS: EOs from the majority of the species tested (S. tortuosum, A. campestris subsp. maritima, O. maritimus, and E. maritimum) were mainly composed by hydrocarbon compounds (sequisterpenes and monoterpenes), showing antiproliferative activity in both 2D and 3D models. EO extracted from S. tortuosum and O. maritimus were identified as having the lowest IC50 values for both cell lines when compared with the other species tested. Furthermore, this antiproliferative activity was associated with increased p21 expression and induction of apoptosis. CONCLUSIONS: The present study suggests that EOs extracted from S. tortuosum and O. maritimus present promising cytotoxic properties. Further evaluation of the extracts and their key components as potential anticancer agents should therefore be explored.

WSB-1 regulates the metastatic potential of hormone receptor negative breast cancer.

BACKGROUND: Metastatic spread is responsible for the majority of cancer-associated deaths. The tumour microenvironment, including hypoxia, is a major driver of metastasis. The aim of this study was to investigate the role of the E3 ligase WSB-1 in breast cancer biology in the context of the hypoxic tumour microenvironment, particularly regarding metastatic spread. METHODS: In this study, WSB-1 expression was evaluated in breast cancer cell lines and patient samples. In silico analyses were used to determine the impact of WSB-1 expression on distant metastasis-free survival (DMFS) in patients, and correlation between WSB1 expression and hypoxia gene expression signatures. The role of WSB-1 on metastasis promotion was evaluated in vitro and in vivo. RESULTS: High WSB1 expression was associated with decreased DMFS in ER-breast cancer and PR-breast cancer patients. Surprisingly, WSB1 expression was not positively correlated with known hypoxic gene expression signatures in patient samples. Our study is the first to show that WSB-1 knockdown led to decreased metastatic potential in breast cancer hormone receptor-negative models in vitro and in vivo. WSB-1 knockdown was associated with decreased metalloproteinase (MMP) activity, vascular endothelial growth factor (VEGF) secretion, and angiogenic potential. CONCLUSIONS: Our data suggests that WSB-1 may be an important regulator of aggressive metastatic disease in hormone receptor-negative breast cancer. WSB-1 could therefore represent a novel regulator and therapeutic target for secondary breast cancer in these patients.

Low molecular weight heparin suppresses tissue factor-mediated cancer cell invasion and migration in vitro.

Elevated expression of tissue factor (TF) has been associated with an increased risk of thrombosis in the majority of cancers. Moreover, treatment of cancer patients with low molecular weight heparin (LMWH) appears to have beneficial effects that reach beyond controlling the immediate hypercoagulable state. In this study, we investigated the influence of the treatment of cancer cells with LMWH (0-2,000 µg/ml) on cell invasiveness and migration in cancer cell lines from five separate tissues; pancreatic, breast, colocarcinoma, ovarian and melanoma. The rate of cell invasion across collagen IV-coated membranes was suppressed in all cell lines tested on incubation with 2,000 µg/ml LMWH, but BxPC-3 and MDA-MB-231 cells also responded to the lowest concentration of 20 µg/ml LMWH. Furthermore, the rate of cell migration was reduced to varying extents in all of the cell lines tested on incubation with 20 µg/ml or higher concentrations of LMWH. The decrease in the rates of invasion and migration also strongly correlated with the reduction in TF protein expression and TF activity in these cells following incubation with LMWH. Moreover, the LMWH-mediated decreases in cellular invasion in the most affected cell lines (BxPC-3 and MDA-MB-231) were restored by transfection of the cells with the mammalian pCMV-XL5-TF expression vector allowing independent overexpression of TF. In conclusion, LMWH appears to suppress the rate of cancer cell invasion and migration in vitro, through a mechanism that is at least in part dependent on the TF protein expression and activity in cancer cells.