Quantitation overcoming Matrix effects of Lipophilic toxins in Mytilus galloprovincialis by liquid chromatography-full scan high resolution mass spectrometry analysis (LC-HR-MS)

The analysis of marine lipophilic toxins in shellfish products still represents a challenging task due to the complexity and diversity of the sample matrix. Liquid chromatography coupled with mass spectrometry (LC-MS) is the technique of choice for accurate quantitative measurements in complex samples. By combining unambiguous identification with the high selectivity of tandem MS, it provides the required high sensitivity and specificity. However, LC-MS is prone to matrix effects (ME) that need to be evaluated during the development and validation of methods. Furthermore, the large sample-to-sample variability, even between samples of the same species and geographic origin, needs a procedure to evaluate and control ME continuously. Here, we analyzed the toxins okadaic acid (OA), dinophysistoxins (DTX-1 and DTX-2), pectenotoxin (PTX-2), yessotoxin (YTX) and azaspiracid-1 (AZA-1). Samples were mussels (Mytilus galloprovincialis), both fresh and processed, and a toxin-free mussel reference material. We developed an accurate mass-extracted ion chromatogram (AM-XIC) based quantitation method using an Orbitrap instrument, evaluated the ME for different types and extracts of mussel samples, characterized the main compounds co-eluting with the targeted molecules and quantified toxins in samples by following a standard addition method (SAM). An AM-XIC based quantitation of lipophilic toxins in mussel samples using high resolution and accuracy full scan profiles (LC-HR-MS) is a good alternative to multi reaction monitoring (MRM) for instruments with HR capabilities. ME depend on the starting sample matrix and the sample preparation. ME are particularly strong for OA and related toxins, showing values below 50% for fresh mussel samples. Results for other toxins (AZA-1, YTX and PTX-2) are between 75% and 110%. ME in unknown matrices can be evaluated by comparing their full scan LC-HR-MS profiles with those of known samples with known ME. ME can be corrected by following SAM with AM-XIC quantitation if necessary.info:eu-repo/semantics/publishedVersio

The Emerging Role of microRNAs in Aquaporin Regulation

Aquaporins (AQPs) are membrane channels widely distributed in human tissues. AQPs are essential for water and energy homeostasis being involved in a broad range of pathophysiological processes such as edema, brain injury, glaucoma, nephrogenic diabetes insipidus, salivary and lacrimal gland dysfunction, cancer, obesity and related metabolic complications. Compelling evidence indicates that AQPs are targets for therapeutic intervention with potential broad application. Nevertheless, efficient AQP modulators have been difficult to find due to either lack of selectivity and stability, or associated toxicity that hamper in vivo studies. MicroRNAs (miRNAs) are naturally occurring small non-coding RNAs that regulate post-transcriptional gene expression and are involved in several diseases. Recent identification of miRNAs as endogenous modulators of AQP expression provides an alternative approach to target these proteins and opens new perspectives for therapeutic applications. This mini-review compiles the current knowledge of miRNA interaction with AQPs highlighting miRNA potential for regulation of AQP-based disorders

The aquaporin-3-inhibiting potential of polyoxotungstates

Polyoxometalates (POMs) are of increasing interest due to their proven anticancer activities. Aquaporins (AQPs) were found to be overexpressed in tumors bringing particular attention to their inhibitors as anticancer drugs. Herein, we report for the first time the ability of polyoxotungstates (POTs), such as of Wells-Dawson P2W18, P2W12, and P2W15, and Preyssler P5W30 structures, to affect aquaporin-3 (AQP3) activity and impair melanoma cell migration. The tested POTs were revealed to inhibit AQP3 function with different effects, with P2W18, P2W12, and P5W30 being the most potent (50% inhibitory concentration (IC50) = 0.8, 2.8, and 3.2 µM), and P2W15 being the weakest (IC50 > 100 µM). The selectivity of P2W18 toward AQP3 was confirmed in yeast cells transformed with human aquaglyceroporins. The effect of P2W12 and P2W18 on melanoma cells that highly express AQP3 revealed an impairment of cell migration between 55% and 65% after 24 h, indicating that the anticancer properties of these compounds may in part be due to the blockage of AQP3-mediated permeability. Altogether, our data revealed that P2W18 strongly affects AQP3 activity and cancer cell growth, unveiling its potential as an anticancer drug against tumors where AQP3 is highly expressed.Fundação para a Ciência e a Tecnologia (FCT) (PTDC/BTM-SAL/28977/2017, PTDC/MED-QUI/31721/2017, UID/DTP/04138/2019, UIDB/04326/2020, SFRH/BD/117586/2016. Austrian Science Fund (FWF) P27534, M2203info:eu-repo/semantics/publishedVersio

Influence of mixtures of acenaphthylene and benzo[a] anthracene on their degradation by Pleurotus ostreatus in sandy soil

Purpose Polycyclic aromatic hydrocarbons (PAHs) are a class of organic compounds commonly found as soil contaminants. Fungal degradation is considered as an environmentally friendly and cost-effective approach to remove PAHs from soil. Acenaphthylene (Ace) and Benzo[a]anthracene (BaA) are two PAHs that can coexist in soils; however, the influence of the presence of each other on their biodegradation has not been studied. The biodegradation of Ace and BaA, alone and in mixtures, by the white rot fungus Pleurotus ostreatus was studied in a sandy soil. Materials and methods Experimental microcosms containing soil spiked with different concentrations of Ace and BaAwere inoculated with P. ostreatus. Initial (t 0) and final (after 15 days of incubation) soil concentrations of Ace and BaA were determined after extraction of the PAHs. Results and discussion P. ostreatus was able to degrade 57.7% of the Ace in soil spiked at 30 mg kg−1 dry soil and 65.8% of Ace in soil spiked at 60 mg kg−1 dry soil. The degradation efficiency of BaA by P. ostreatus was 86.7 and 77.4% in soil spiked with Ace at 30 and 60 mg kg−1 dry soil, respectively. After 15 days of incubation, there were no significant differences in Ace concentration between soil spiked with Ace and soil spiked with Ace + BaA, irrespective of the initial soil concentration of both PAHs. There were also no differences in BaA concentration between soil spiked with BaA and soil spiked with BaA + Ace. Conclusions The results indicate that the fungal degradation of Ace and BaA was not influenced by the presence of each other’s PAH in sandy soil. Bioremediation of soils contaminated with Ace and BaA using P. ostreatus is a promising approach to eliminate these PAHs from the environment

Factors associated with self-reported discrimination against men who have sex with men in Brazil

OBJECTIVE: To estimate self-reported discrimination due to sexual orientation among men who have sex with men (MSM) in Brazil and to analyze associated factors. METHODS: A cross-sectional study of 3,859 MSM recruited in 2008–2009 with respondent driven sampling. Data collection conducted in health centers in 10 Brazilian cities. A face-to-face questionnaire was used and rapid HIV and syphilis tests conducted. Aggregated data were weighted and adjusted odds ratio estimated to measure the association between selected factors and self-reported discrimination due to sexual orientation. RESULTS: The sample was predominantly young, eight plus years of schooling, pardo (brown), single, low-income, and identified themselves as gay or homosexual. The prevalence of self-reported discrimination due to sexual orientation was 27.7% (95%CI 26.2–29.1). Discrimination was independently associated with: age < 30 years, more years of schooling,community involvement and support, history of sexual and physical violence, suicidal thoughts, and unprotected receptive anal intercourse. CONCLUSIONS: The prevalence of self-reported discrimination among MSM in Brazil is high. These results challenge the assumptions that MSM-specific prevention and support programs are not required or that health professionals do not need special training to address MSM needs

Glutamine and cystine-enriched diets modulate aquaporins gene expression in the small intestine of piglets

Research Areas: Science & Technology ; Other TopicsABSTRACT - The regulation of glycerol permeability in the gastrointestinal tract is crucial to control fat deposition, lipolysis and gluconeogenesis. Knowing that the amino acid glutamine is a physiological regulator of gluconeogenesis, whereas cystine promotes adiposity, herein we investigated the effects of dietary supplementation with glutamine and cystine on the serum biochemical parameters of piglets fed on amino acid-enriched diets, as well as on the transcriptional profile of membrane water and glycerol channels aquaporins (AQPs) in the ileum portion of the small intestine and its impact on intestinal permeability. Twenty male piglets with an initial body weight of 8.8 ± 0.89 kg were allocated to four dietary treatments (n = 5) and received, during a four week-period, a basal diet without supplementation (control) or supplemented with 8 kg/ton of glutamine (Gln), cystine (Cys) or the combination of the two amino acids in equal proportions (Gln + Cys). Most biochemical parameters were found improved in piglets fed Gln and Cys diet. mRNA levels of AQP3 were found predominant over the others. Both amino acids, individually or combined, were responsible for a consistent downregulation of AQP1, AQP7 and AQP10, without impacting on water permeability. Conversely, Cys enriched diet upregulated AQP3 enhancing basolateral membranes glycerol permeability and downregulating glycerol kinase (GK) of intestinal cells. Altogether, our data reveal that amino acids dietary supplementation can modulate intestinal AQPs expression and unveil AQP3 as a promising target for adipogenesis regulation.info:eu-repo/semantics/publishedVersio

Application of a purge-and-trap system for fixed-NH4+ determination by matrix interference free ion chromatography in oil reservoir rocks

A method for fixed-NH4+ determination using a system immersed into an ultrasonic bath for NH3 extraction and ion chromatography for NH4+ determination is proposed. The method is faster, simpler and more sensible (LOD = 4.4 mg g-1 NH4+) than the classical Kjeldahl method

Montrichardia linifera (Araceae) biological potential, phytochemical prospection and polyphenol content

Montrichardia linifera is commonly known in Brazil as "aninga." In November 2008 and August 2009, we collected plant samples and extracted leaves and stems using hexane or dichloromethane and methanol to evaluate its biological and chemical potential, phytochemical profile and polyphenol content. Differences in the chemical composition of the collections were evident. In the first collection, the apolar extracts were the richest in metabolite composition while the polar extracts were the richest in the second collection. In general, extracts from Aug/2009 had higher antioxidant and cytotoxic activities than extracts from Nov/2008 and the stem and leaf extracts from the Aug/2009 collection showed the highest polyphenol content. As an insecticide against Sitophilus zeamais, the hexane extract of the leaves (Aug/2009) presented concentration-dependent insecticidal activity. Only the methanolic extract of stems and leaves (Aug/2009) presented a small inhibition halo (9 and 7 mm, respectively) against Aeromonas hydrophila when evaluating its antibacterial potential. The study demonstrates that, collection period is an important factor to consider in phytochemical and biological activity studies of M. linifera

An atherogenic diet disturbs aquaporin 5 expression in liver and adipocyte tissues of apolipoprotein e-deficient mice: new insights into an old model of experimental atherosclerosis

The dysfunction of vascular endothelial cells is profoundly implicated in the pathogenesis of atherosclerosis and cardiovascular disease, the global leading cause of death. Aquaporins (AQPs) are membrane channels that facilitate water and glycerol transport across cellular membranes re-cently implicated in the homeostasis of the cardiovascular system. Apolipoprotein-E deficient (apoE−/−) mice are a common model to study the progression of atherosclerosis. Nevertheless, the pattern of expression of AQPs in this atheroprone model is poorly characterized. In this study, apoE−/− mice were fed an atherogenic high-fat (HF) or a control diet. Plasma was collected at multiple time points to assess metabolic disturbances. At the endpoint, the aortic atherosclerotic burden was quantified using high field magnetic resonance imaging. Moreover, the transcriptional levels of several AQP isoforms were evaluated in the liver, white adipocyte tissue (WAT), and brown adipocyte tissue (BAT). The results revealed that HF-fed mice, when compared to controls, presented an ex-acerbated systemic inflammation and atherosclerotic phenotype, with no major differences in systemic methylation status, circulating amino acids, or plasma total glutathione. Moreover, an over-expression of the isoform AQP5 was detected in all studied tissues from HF-fed mice when compared to controls. These results suggest a novel role for AQP5 on diet-induced atherosclerosis that warrants further investigation