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Consumers are increasingly concerned about cosmetic ingredients' origin, looking more than ever to sustainable and greener formulations. The Natural Park of Montesinho, located in Portugal, is characterized by an enormous fauna and flora diversity. Among them, beeswax and strawberry trees ( Arbutus unedo ) have attracted the cosmetic researchers' interest due to their bioactive compounds' richness, particularly fatty acids and phenolic compounds. The main goal of this study was to develop an innovative cosmetic product with antioxidant properties composed by both matrices. Briefly, samples were obtained in the Natural Park of Montesinho in October 2022. Beeswax was analysed for lipid profile and contaminants, while extracts were obtained from fruits by ultrasound-assisted extraction (UAE) using water as a solvent. The effect of extraction time (15-90 min) was studied on the total phenolic content (TPC), in vitro antioxidant/antiradical activity, and reactive oxygen species (ROS) scavenging capacity. The beeswax lipid profile presented a high incidence of palmitic, oleic, and linoleic acids. The extract obtained at 60 min presented the highest TPC (30.27 mg GAE/g dw) and antioxidant/antiradical activities (ABTS = 30.36 mg AAE/g dw; DPPH = 43.83 mg TE/g dw; FRAP = 415.61 µmol FSE/g dw). An IC 50 of 19.78 µg/mL was achieved for the hypochlorous acid, while for superoxide radical and peroxyl radical the IC 50 were, respectively, 90.51 µg/mL and 0.19 µmol TE/mg dw. The phytochemical profile revealed a high content of gallic acid, and catechin and its derivatives. The hydrophilic cream developed revealed ideal technological parameters, particularly its stability.

Sargassum muticum (SM) poses a serious environmental issue since it is a fast-expanding invasive species occupying key areas of the European shoreline, disrupting the autochthonous algae species, and disturbing the ecosystem. This problem has concerned the general population and the scientific community. Nevertheless, as macroalgae are recognized as a source of bioactive molecules, the abundance of SM presents an opportunity as a raw material. In this work, response surface methodology (RSM) was applied as a tool for the optimization of the extraction of bioactive compounds from SM by microwave-assisted extraction (MAE). Five different parameters were used as target functions: yield, total phenolic content (TPC); and the antioxidant measurements of 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity (DPPH), 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS), and β-carotene bleaching (BC). After the optimal extraction conditions were determined (time = 14.00 min; pressure = 11.03 bar; ethanol = 33.31%), the chemical composition and bioactivity of the optimum extract was evaluated to appraise its antioxidant capability to scavenge reactive species and as a potential antibacterial, antidiabetic, antiproliferation, and neuroprotective agent. The results lead to the conclusion that MAE crude extract has bioactive properties, being especially active as an antiproliferation agent and as a nitric oxide and superoxide radical scavenger.

Polycyclic aromatic hydrocarbons (PAHs) are widespread across the environment and humans are unavoidably and constantly exposed to them. As lipophilic contaminants, these substances tend to accumulate in fatty tissues as adipose tissue and exposure to these endocrine disruptors has been associated with severe health hazards including prevalence and incidence of obesity. Previous studies have shown significantly higher concentrations of PAHs in adipose tissue compared to other human samples, such as urine and plasma, which are typically used for PAHs assessment. Therefore, conducting biomonitoring studies in adipose tissue is essential, although such studies are currently limited. In this study, the concentrations of 18 PAHs were measured in subcutaneous (scAT) and visceral adipose tissue (vAT) of 188 Portuguese obese females by high performance liquid chromatography (HPLC). The obtained results were then associated with the patient's data namely: 13 clinical, 4 social, and 42 biochemical parameters. Seventeen PAHs were present, at least, in one sample of both scAT and vAT, most of them with detection frequencies higher than 80%. Indeno [1,2,3-cd]pyrene (InP) was the only PAH never detected. Overall higher concentrations of PAHs were observed in scAT. Median concentrations of ∑PAHs were 32.2 ± 10.0 ng/g in scAT and 24.6 ± 10.0 ng/g in vAT. Thirty-six significant associations (7 with social, 18 with clinical, and 11 with biochemical parameters), including 21 Spearman's correlations were identified (12 positive and 9 negative correlations). Indicating the potential effects of PAHs on various parameters such as obesity evolution, body fat, number of adipocytes, total cholesterol, alkaline phosphatase, macrominerals, uric acid, sedimentation velocity, and luteinizing hormone. This study underscores the significance of biomonitoring PAH levels in adipose tissue and their potential effects on metabolic health. Further research is essential to fully comprehend the metabolic implications of PAHs in the human body and to develop strategies for obesity prevention and treatment., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Olive leaves are consumed as an extract or as a whole herbal powder with several potential therapeutic benefits attributed to polyphenols, tocopherol's isomers, and flavonoids, among others. This study assessed the potential variance in the functional features presented by olive leaves from three different Portuguese cultivars-Cobrançosa, Madural, and Verdeal-randomly mix-cultivated in the geographical area of Vale de Salgueiros. Inorganic analysis determined their mineral profiles while an organic analysis measured their total phenolic and flavonoid content, and scanned their phenolic and tocopherol and fatty acid composition. The extracts' biological activity was tested by determining their antimicrobial and antioxidant power as well as their ability to inhibit acetylcholinesterase, butyrylcholinesterase, MAO-A/B, and angiotensin-I-converting enzyme. The inorganic profiles showed them to be an inexpensive source able to address different mineral deficiencies. All cultivars appear to have potential for use as possible antioxidants and future alternative antibiotics against some multidrug-resistant microorganisms, with caution regarding the arsenic content in the Verdeal cultivar. Madural's extract displayed properties to be considered a natural multitarget treatment for Alzheimer's and Parkinson's diseases, depression, and cardiometabolic and dual activity for blood pressure modulation. This work indicates that randomly cultivating different cultivars significantly modifies the leaves' composition while keeping their multifaceted therapeutic value.

Cardiovascular diseases are among the major causes of mortality and morbidity. Warfarin is often prescribed for these disorders, an anticoagulant with inter and intra-dosage variability dose required to achieve the target international normalized ratio. Warfarin presents a narrow therapeutic index, and due to its variability, it can often be associated with the risk of hemorrhage, or in other patients, thromboembolism. Single-nucleotide polymorphisms are included in the causes that contribute to this variability. The Cytochrome P450 (CYP) 2C9*3 genetic polymorphism modifies its enzymatic activity, and hence warfarin's plasmatic concentration. Thus, the need for a selective, rapid, low-cost, and real-time detection device is crucial before prescribing warfarin. In this work, a disposable electrochemical DNA-based biosensor capable of detecting CYP2C9*3 polymorphism was developed. By analyzing genomic databases, two specific 78 base pairs DNA probes; one with the wild-type adenine (Target-A) and another with the cytosine (Target-C) single-nucleotide genetic variation were designed. The biosensor implied the immobilization on screen-printed gold electrodes of a self-assembled monolayer composed by mercaptohexanol and a linear CYP2C9*3 DNA-capture probe. To improve the selectivity and avoid secondary structures a sandwich format of the CYP2C9*3 allele was designed using complementary fluorescein isothiocyanate-labeled signaling DNA probe and enzymatic amplification of the electrochemical signal. Chronoamperometric measurements were performed at a range of 0.015-1.00 nM for both DNA targets achieving limit of detection of 42 p.m. The developed DNA-based biosensor was able to discriminate between the two synthetic target DNA targets, as well as the targeted denatured genomic DNA, extracted from volunteers genotyped as non-variant homozygous (A/A) and heterozygous (A/C) of the CYP2C9*3 polymorphism., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Exposure to environmental chemicals during developmental stages can result in several adverse outcomes. In this study, the exposure of Portuguese children to Cu, Co, I, Mo, Mn, Ni, As, Sb, Cd, Pb, Sn and Tl was evaluated through the analysis of first morning urine through ICP-MS. Furthermore, we attempted to determine possible exposure predictors. The study sample consisted of 54% girls and 46% boys, with a median age of 10 years; 61% were overweight/obese and were put on a nutritionally oriented diet. For I, half of the population was probably in deficiency status. The median urinary concentrations (μg/L) were Cu 21.9, Mo 54.6, Co 0.76, Mn 2.1, Ni 4.74, As 37.9, Sb 0.09, Cd 0.29, Pb 0.94, Sn 0.45, Tl 0.39 and I 125.5. The region was a significant predictor for Cu, Co, Ni, As and Tl. Children living in an urban area had higher urinary levels, except for Co and Ni. Age was a significant predictor for Cu, I, Mo, Mn, Ni, Sb, Cd and Sn with urinary levels of these elements decreasing with age. No sex-related differences were observed. Diet and weight group were predictors for urinary Cu, Mn, Ni, Sb and As. Significant differences were observed between the diet/weight groups for Cu, Ni, Sb and As, with the healthy diet group presenting higher values.

Polycyclic aromatic hydrocarbons (PAHs) are environmentally persistent organic pollutants formed during incomplete combustion and pyrolysis processes. Humans are continuously exposed to PAHs which are linked to severe health effects such as diabetes, cancer, infertility, and poor foetal development, amongst others. PAHs are lipophilic compounds prone to accumulating in adipose tissue. Even though adipose tissue is the ideal matrix to assess over time accumulation of lipophilic pollutants, only a few analytical methods have been developed for this matrix. Aiming to reduce the existent gap, a method for the extraction of PAHs from adipose tissue samples using ultrasound-assisted extraction (UAE) was developed. The behaviour of PAHs (retention, adsorption, and volatilization) over several steps of the analytical procedure was studied. Validation tests were performed on the optimized method. PAHs were quantified using a high performance liquid chromatography (HPLC) system equipped with a photodiode array (PDA) and fluorescence (FLD) detector inline. The method achieved a low matrix effect and presents low method detection (MDL) and quantification (MQL) limits, showing suitability for a selective and sensitive determination of PAHs in adipose tissue. The extraction is performed with 0.4 g of adipose tissue and 6 mL of n -hexane and it does not require clean-up afterwards. Additionally, an Eco-Scale score of 74 and an Analytical GREEnness score of 0.66 were obtained. The method achieved is effective, simpler, greener, and easy to perform, being an alternative to conventional extraction methods. Furthermore, this method can be used as a multi-analyte methodology since it has been previously validated by the authors for the analysis of other lipophilic compounds. Naphthalene (Naph), acenaphthene (Ace), fluorene (Flu), phenanthrene (Phe), anthracene (Ant), fluoranthene (Fln), pyrene (Pyr) and benzo[ k ]fluoranthene (B[ k ]Ft) were found in all the tested adipose tissue samples.

A new application of graphene-type materials as an alternative cleanup sorbent in a quick, easy, cheap, effective, rugged, and safe (QuEChERS) procedure combined with GC-ECD/GC-MS/GC-MS/MS detection was successfully used for the simultaneous analysis of 12 brominated flame retardants in Capsicum cultivar samples. The chemical, structural, and morphological properties of the graphene-type materials were evaluated. The materials exhibited good adsorption capability of matrix interferents without compromising the extraction efficiency of target analytes when compared with other cleanups using commercial sorbents. Under optimal conditions, excellent recoveries were obtained, ranging from 90 to 108% with relative standard deviations of <14%. The developed method showed good linearity with a correlation coefficient above 0.9927, and the limits of quantification were in the range of 0.35-0.82 μg/kg. The developed QuEChERS procedure using reduced graphite oxide (rGO) combined with GC/MS was successfully applied in 20 samples, and the pentabromotoluene residues were quantified in two samples.

The production and consumption of shrimp species create massive amounts of shrimp bio-waste. In this study, shrimp shell waste from Palaemon serratus and Palaemon varians from the Portuguese coast was characterized. Regarding the antioxidant capacity, the obtained values were between 4.7 and 10.4 mg gallic acid equivalents (GAE)/g dry weight (dw) for Total phenolic content (TPC); 3 and 7 mg ascorbic acid equivalents (AAE)/g dw for Ferric reducing antioxidant power assay (FRAP); 0.4 and 1.2 mg Trolox equivalent (TE)/g dw for 2,2-diphenyl-1-picryl-hydrazyl-hydrate free radical scavenging (DPPH • ); 4 and 11 mg TE/g dw for 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) radical scavenging activity (ABTS •+ ); and 72 and 130 mg TE/g dw for Oxygen radical absorbance capacity (ORAC). For the antimicrobial activity, shrimp shell waste from P. varians formed inhibition zones between 14 and 23 mm. Total carotenoid content values were in the range of 28 and 134 μg/g dw, and according to their HPLC-PAD profile, β-carotene and astaxanthin contents were between 0.3 and 7.6 μg/g dw and 1.1 and 26.1 μg/g dw, respectively. These studies are critical to recognizing the potential added value of shrimp shell waste as possible colorants and preservatives with antioxidant protection capacity to be used in the food industry.

The aims of this study were to characterize the exposure of pregnant women living in Portugal to 3-phenoxybenzoic acid (3-PBA) and to evaluate the association of this exposure with maternal outcomes and newborn anthropometric measures. We also aimed to compare exposure in summer with exposure in winter. Pregnant women attending ultrasound scans from April 2018 to April 2019 at a central hospital in Porto, Portugal, were invited to participate. Inclusion criteria were: gestational week between 10 and 13, confirmed fetal vitality, and a signature of informed consent. 3-PBA was measured in spot urine samples by gas chromatography with mass spectrometry (GC-MS). The median 3-PBA concentration was 0.263 (0.167; 0.458) µg/g creatinine ( n = 145). 3-PBA excretion was negatively associated with maternal pre-pregnancy body mass index (BMI) ( p = 0.049), and it was higher during the summer when compared to winter ( p < 0.001). The frequency of fish or yogurt consumption was associated positively with 3-PBA excretion, particularly during the winter ( p = 0.002 and p = 0.015, respectively), when environmental exposure is low. Moreover, 3-PBA was associated with levothyroxine use ( p = 0.01), a proxy for hypothyroidism, which could be due to a putative 3-PBA-thyroid hormone antagonistic effect. 3-PBA levels were not associated with the anthropometric measures of the newborn. In conclusion, pregnant women living in Portugal are exposed to 3-PBA, particularly during summer, and this exposure may be associated with maternal clinical features.

Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous global pollutants that are lipophilic, bioaccumulative and can be both endocrine disruptors and carcinogens. In this study, we measured 18 PAHs among decapod shrimp samples (Palaemon spp.) collected during the spring and autumn from multiple locations along the Portuguese coastal, including local markets and aquaculture farms. ΣPAH concentrations in samples ranged from 3.14 to 320.48 ng/g (ww) with the highest value in this range detected in a sample provided from an aquaculture facility. Fifteen of the 18 scanned PAHs were present in shrimp samples. However, detectable concentrations (0.07 ng/g ww) of the carcinogenic benzo-a-pyrene occurred for a single sample only and approached method detection limits. No significant differences in shrimp ΣPAH concentrations were evident between the spring and autumn seasons. Target hazard quotient estimation suggested low probability of adverse health effects to consumers through consumption of shrimp from the study collection locations., (© 2022. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Brominated flame retardants (BFRs) are chemicals employed to lower the flammability of several objects. These endocrine disruptor chemicals are lipophilic and persistent in the environment. Due to these characteristics some have been restricted or banned by the European Union, and replaced by several new chemicals, the novel BFRs (NBFRs). BFRs are widely detected in human samples, such as adipose tissue and some were linked with altered thyroid hormone levels, liver toxicity, diabetes and metabolic syndrome in humans. However, the disturbance in lipid metabolism caused by BFRs with emphases to NBFRs remains poorly understood. In this study, we used a pre-adipocyte (3T3-L1) cell line and a hepatocyte (HepG2) cell line to investigate the possible lipid metabolism disruption caused by four BFRs: hexabromobenzene (HBB), pentabromotoluene (PBT), 2-ethylhexyl-2,3,4,5-tetrabromobenzoate (TBB) and hexabromocyclododecane (HBCD). For that purpose, proliferation and Oil Red O assays, as well as, medium fatty acids profile evaluation using Gas chromatography and RNA extraction for quantitative RT-PCR assays were performed. We detected a significant reduction in the proliferation of preadipocytes and an increased lipid accumulation during differentiation caused by HBB. This BFR also lead to a significant increased expression of IL-1β and decreased expression of PGC-1α and adiponectin. Nevertheless, PBT, TBB and HBCD show to increase lipid accumulation in hepatocytes. PBT also display a significant increase of PPARγ gene expression. Lipid accumulation in the cells can occur by diverse mechanisms depending on the BFR. These results highlight the importance of endocrine disruptor compounds in obesity etiopathogeny., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Humans are exposed every day to assorted environmental pollutants namely, polychlorinated biphenyls (PCB), organochlorine pesticides (OCP), brominated flame-retardants (BFR), polycyclic aromatic hydrocarbons (PAH), synthetic musks, heavy metals (e.g. cadmium) and plastic additives (e.g. bisphenol A, BPA). Besides environmental persistence, biomagnification and bioaccumulative properties, these pollutants are classified as endocrine disruptors (EDs), metabolic disruptors, neurologic disruptors. These compounds affect the normal function of several mechanisms in the human body being linked to human health issues as cancer development (e.g. breast, uterine and endometrial), miscarriage, birth defects, premature delivery and infertility. In order to prevent future health issue of women and possible progeny, the assessment of EDs accumulated is essential, particularly in adipose tissue. These samples have been referred as the ideal matrix to establish over time accumulation and long-term exposure of persistent and non-persistent lipophilic EDs. However, the invasive sample collection procedure and methodology processing discourages the usage of this matrix for biomonitoring studies. In this review, a Web of Science search without any publishing year restriction on the analysis in adipose tissue of PCB, OCP, BPA, cadmium, BFR and synthetic musks was performed. A total of 313 studies were found, 158 were European studies from which the studies with data on women EDs accumulation were selected for detailed analysis (n = 90). The results were structured and presented in accordance with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. The present paper is an overview on the existent EDs analytical methods and levels accumulated in women adipose tissue, with the correspondent health implications across Europe. The limits of detection and quantification were compared and a discussion with results obtained about the presence of ED was performed., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Efforts have been made to find natural, highly nutritious alternatives to replace fish meal (FM) and fish oil (FO), which can simultaneously promote fish health and improve the nutritional quality of filets for human consumption. This study evaluated the impact of biofortified diets containing microalgae (as replacement for FM and FO), macroalgae (as natural source of iodine) and selenised yeast (organic source of selenium) on gilthead seabream growth, nutrient utilization, tissue composition and gene expression. A control diet (CTRL) with 15% FM and 5.5% FO was compared with three experimental diets (AD1, AD2, and AD3), where a microalgae blend ( Chlorella sp., Tetraselmis sp., and DHA-rich Schizochytrium sp.) replaced 33% of FM. Diet AD1 contained 20% less FO. Diets were supplemented with Laminaria digitata (0.4% AD1 and AD2; 0.8% AD3) and selenised yeast (0.02% AD1 and AD2; 0.04% AD3). After feeding the experimental diets for 12 weeks, growth was similar in fish fed AD1, AD2, and CTRL, indicating that microalgae meal can partially replace both FM and FO in diets for seabream. But AD3 suppressed fish growth, suggesting that L. digitata and selenised yeast supplementation should be kept under 0.8 and 0.04%, respectively. Despite lower lipid intake and decreased PUFAs bioavailability in fish fed AD3, compared to CTRL, hepatic elovl5 was upregulated resulting in a significant increase of muscle EPA + DHA. Indeed, filets of fish fed AD2 and AD3 provided the highest EPA + DHA contents (0.7 g 100 g -1 ), that are well above the minimum recommended values for human consumption. Fish consuming the AD diets had a higher retention and gain of selenium, while iodine gain remained similar among diets. Upregulation of selenoproteins ( gpx1 , selk , and dio2 ) was observed in liver of fish fed AD1, but diets had limited impact on fish antioxidant status. Overall, results indicate that the tested microalgae are good sources of protein and lipids, with their LC-PUFAs being effectively accumulated in seabream muscle. Selenised yeast is a good fortification vehicle to increase selenium levels in fish, but efforts should be placed to find new strategies to fortify fish in iodine., Competing Interests: JD was employed by the company Sparos Lda. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Ferreira, Ribeiro, Ribeiro, Barata, Domingues, Sousa, Soares, Marques, Pousão-Ferreira, Dias, Castro, Marques, Nunes and Valente.)

Three sampling campaigns were performed in the Lis River (Leiria, Portugal) in February of 2018, November of 2018, and May of 2019. River water and wastewater (influent and effluent) samples of two wastewater treatment plants were target of the study. A total of 25 samples were collected and 50 pesticides were monitored, including organochlorines, triazines, pyrethroids, and organophosphorus, among others. Most of the detected pesticides were insecticides and mainly organochlorines. Concentrations between 1.29 and 2134 ng/L were found. Aldrin, γ-HCH, and cypermethrin were detected in some samples in μg/L, being γ-HCH the pesticide most frequently detected with concentration in μg/L level. The pesticides with the highest detection frequency were (i) cypermethrin, HCB, methoxychlor, and ζ-HCH in river waters; (ii) isoproturon, cypermethrin, methoxychlor, pyrimethanil, γ-HCH, dieldrin, diuron, α-HCH, and α-endosulfan in effluents; and (iii) diuron and isoproturon in influents. The detection of the organochlorides and their degradation products is a consequence of their persistence in the environment, as their usage has long been prohibited in the European Union. Pesticides were grouped by their types in herbicides, insecticides, or fungicides and the detection and concentration for each type were discussed with the climatic conditions. Pesticide toxicity index was determined in the samples collected in the river., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

The aim of this study was to evaluate the influence of the fishing location on yellowfin tuna's (YFT; Thunnus albacares ) white muscle total lipid (TL) content and fatty acid profile. The comparison included 45 YFT loins, equally divided between the Atlantic, Indian and Pacific Oceans. The ocean had no significant influence on YFT TL content, total PUFA and total n-3 PUFA ( p > 0.05), averaging 1.09 g/100 g of muscle, 229.2 and 192.8 mg/100 g of muscle, respectively. On the other hand, the YFT harvested on the Indian Ocean displayed significantly higher contents of both SFA and MUFA totals than the Atlantic Ocean counterparts ( p < 0.05), while the YFT harvested in the Pacific Ocean presented intermediate values, not differing significantly from the other two origins. The YFT from the Indian and Pacific oceans held twice the n-6 PUFA content recorded in the Atlantic YFT (44.1 versus 21.1 mg/100 g of muscle). Considering the recommended daily intake of EPA plus DHA is 250 mg, the consumption of 100 g of YFT from the Atlantic, Indian and Pacific Oceans would provide 149.2 mg, 191.8 mg or 229.6 mg of EPA plus DHA, representing 59.7%, 76.7% or 91.8% of the recommended daily intake, respectively.

The development of coastal regions has contributed to the intensification of environmental contamination, which can accumulate in aquatic biota, such as shrimps. These crustaceans, besides being delicious and being a good source of nutrients, can also accumulate environmental pollutants. Amongst others, these include organochlorine pesticides (OCPs), organophosphorus pesticides (OPPs), brominated flame retardants (BFRs), polychlorinated biphenyls (PCBs) and synthetic musks (SMs). These pollutants, classified as endocrine disruptors, are related to adverse effects in humans and since one of the major routes of exposition is ingestion, this is a cause for concern regarding their presence in food. The aim of the present study was to quantify the presence of environmental pollutants in shrimp samples and in the water from their habitat along the northwest Portuguese coast. In seawater samples, only two OCPs (lindane and DDD) and one BFR (BTBPE) were detected, and in shrimp samples, one OCP (DDD) and three SMs (HHCB, AHTN and ketone) were found. Bioaccumulation and the risk assessment of dietary exposure of SMs in shrimp samples were investigated. It was observed that all shrimp samples analyzed significantly presented bioaccumulation of the three SMs found. Concentrations of SMs detected in shrimp samples do not present a health risk for the adult Portuguese population.

This study aimed to devise innovative, tailor-made, appealing, tasty and semi-industrialized dishes, using sustainable and under-utilized seafood species (bib, common dab, common carp, blue mussel and blue whiting), that can meet the specific nutritional and functional needs of children (8-10-years), pregnant women (20-40-years) and seniors (≥60-years). Hence, contests were organised among cooking schools from 6 European countries and the best recipes/dishes were reformulated, semi-industrially produced and chemically and microbiologically evaluated. The dishes intended for: (i) children and pregnant women had EPA + DHA and I levels that reached the target quantities, supporting the claim as "high in I"; and (ii) seniors were "high in protein" (24.8%-Soup&#95;S and 34.0%-Balls&#95;S of the energy was provided by proteins), "high in vitamin B12", and had Na contents (≤0.4%) below the defined limit. All dishes reached the vitamin D target value. Sausages&#95;C, Roulade&#95;P, Fillet&#95;P and Balls&#95;S had a well-balanced protein/fat ratio. Roulade&#95;P presented the highest n-3 PUFA/n-6 PUFA ratio (3.3), while Sausages&#95;C the lowest SFA/UNS ratio (0.2). Dishes were considered safe based on different parameters (e.g. Hg-T, PBDEs, Escherichia coli). All represent dietary sources contributing to meet the reference intakes of target nutrients (33->100%), providing valuable options to overcome nutritional and functional imbalances of the three groups., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Algae are an underexploited source of natural bioactive compounds in Western countries, so an increasing interest in the valorization of these marine organisms has emerged in recent years. In this work, the effect of extracting solvent on the extraction yield, phenolic content, antioxidant capacity, and antimicrobial activity of nine brown macroalgae species ( Ascophyllum nodosum , Himanthalia elongata , Undaria pinnatifida , Pelvetia canaliculata , Saccharina latissima , Bifurcaria bifurcata , Laminaria ochroleuca , Sargassum muticum , and Fucus spiralis ) was assessed. Total phenolic content (TPC) and the antioxidant properties of extracts by different assays: radical scavenging activity (DPPH-RSA) and ferric reducing antioxidant power (FRAP) were performed. The antimicrobial activity of extracts was studied against six different foodborne microorganisms: Staphylococcus aureus , Staphylococcus epidermidis , Bacillus cereus , Escherichia coli , Salmonella enteritidis , and Pseudomonas aeruginosa. The highest extraction yield was achieved in ethanolic extracts. However, the highest TPC and FRAP values were obtained on the ethyl acetate extracts, especially from A. nodosum . Concerning algal species, the highest TPC and FRAP values were found in A. nodosum , while the highest DPPH-RSA values were achieved in the hexane extracts of B. bifurcata . The antimicrobial activity of algal extracts varied according to the solvent and alga selected, suggesting the species- and solvent-dependent behavior of this property, with B. bifurcata extracts showing the highest results for a wide range of bacteria. Our results provide insight on the characterization of widespread brown algae in the coasts of the North-Western region of the Iberian Peninsula, reflecting multiple health-enhancing properties which may lead to their exploitation in food, pharmacological, and cosmetic industries.

The total lipid content and lipidic profile of seaweeds harvested in the North Coast and purchased in Portugal were determined in this paper. The amount of total lipids in the different species of seaweeds varied between 0.7 ± 0.1% ( Chondrus crispus ) and 3.8 ± 0.6% ( Ulva spp.). Regarding the fatty acid content, polyunsaturated fatty acids (PUFA) ranged between 0-35%, with Ulva spp. presenting the highest amount; monounsaturated fatty acids (MUFA) varied between 19 and 67%; and saturated fatty acids (SFA) were predominant in C. crispus (45-78%) and Gracilaria spp. (36-79%). Concerning the nutritional indices, the atherogenicity index (AI) was between 0.4-3.2, the thrombogenicity index (TI) ranged from 0.04 to 1.95, except for Gracilaria spp., which had a TI of 7.6, and the hypocholesterolemic/hypercholesterolemic ratio (HH) values ranged between 0.88-4.21, except for Gracilaria spp., which exhibited values between 0.22-9.26. The n6/n3 ratio was below 1 for most of the species evaluated, except for Ascophyllum nodosum , which presented a higher value, although below 2. Considering the PUFA/SFA ratio, seaweeds presented values between 0.11-1.02. The polycyclic aromatic hydrocarbons (PAHs) and aliphatic hydrocarbons (AHCs) contamination of seaweeds under study was also quantified, the values found being much lower than the maximum levels recommended for foodstuff.

Synthetic musks and organophosphorus pesticides represent a potential risk to the human health since exposure can lead to distinct types of carcinogenesis and endocrine disorders. These are lipophilic compounds as such, prone to deposit and persist in fat tissues, mainly in adipose tissue. Very few studies have reported on the occurrence and accumulation profile of these contaminants in human adipose tissue. Analytical methods for the detection and quantification of synthetic musks and organophosphorus pesticides in adipose tissue are lacking. In this study, the efficacy of different extraction with ultrasonic homogenizer and dispersive solid-phase extraction (d-SPE) clean-up methods were evaluated in human adipose tissue. The relative sample clean-up was assessed by measurement of total lipid content. The quantification of four synthetic musks and six organophosphorus pesticides were performed by gas chromatography (GC) mass spectrometry (MS) and flame photometric detection (FPD), respectively. The d-SPE clean-up with 50 mg PSA, 150 mg MgSO 4 , 100 mg C18EC and 50 mg Z-Sep provided the most effective clean-up, removing the greatest amount of interfering substances including lipids and simultaneously ensuring good chromatographic separation and recoveries. Method detection limits were between 4 to 9 ng/g for synthetic musk and 1 to 7 ng/g for organophosphorus pesticides in adipose tissue. The proposed method was applied to adipose tissue of obese patients and positive samples were confirmed with GC tandem mass spectrometry. Galaxolide was found in all the samples tested with concentrations ranging from 0.08 to 0.5 μg/g of adipose tissue. No other synthetic musk studied was detected. Organophosphorus pesticides were not found in the analysed samples. The developed analytical procedures were successful and can easily be applied to biomonitoring these compounds in human adipose tissue., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Bone graft infections are serious complications in orthopaedics and the growing resistance to antibiotics is increasing the need for antibacterial strategies. The use of magnesium oxide (MgO) is an interesting alternative since it possesses broad-spectrum antibacterial activity. Additionally, magnesium ions also play a role in bone regeneration, which makes MgO more appealing than other metal oxides. Therefore, a bone substitute composed of hydroxyapatite and MgO (HAp/MgO) spherical granules was developed using different sintering heat-treatment cycles to optimize its features. Depending on the sintering temperature, HAp/MgO spherical granules exhibited distinct surface topographies, mechanical strength and degradation profiles, that influenced the in vitro antibacterial activity and cytocompatibility. A proper balance between antibacterial activity and cytocompatibility was achieved with HAp/MgO spherical granules sintered at 1100 ºC. The presence of MgO in these granules was able to significantly reduce bacterial proliferation and simultaneously provide a suitable environment for osteoblasts growth. The angiogenic and inflammation potentials were also assessed using the in vivo chicken embryo chorioallantoic membrane (CAM) model and the spherical granules containing MgO stimulated angiogenesis without increasing inflammation. The outcomes of this study evidence a dual effect of MgO for bone regenerative applications making this material a promising antibacterial bone substitute.

The search for food resources is a constant in human history. Nowadays, the search for natural and safe food supplies is of foremost importance. Accordingly, there is a renewed interest in eco-friendly and natural products for substitution of synthetic additives. In addition, microbial contamination of food products during their obtaining and distribution processes is still a sanitary issue, and an important target for the food industry is to avoid food contamination and its related foodborne illnesses. These diseases are fundamentally caused by certain microorganisms listed in this review and classified according to their Gram negative or positive character. Algae have proven to possess high nutritional value and a wide variety of biological properties due to their content in active compounds. Among these capabilities, macroalgae are recognized for having antimicrobial properties. Thus, the present paper revises the actual knowledge of microbial contaminants in the food industry and proposes antimicrobial algal compounds against those pathogenic bacteria responsible for food contamination as valuable molecules for its growth inhibition. The capacity of algae extracts to inhibit some major food pathogen growth was assessed. Moreover, the main applications of these compounds in the food industry were discussed while considering their favorable effects in terms of food safety and quality control.

Shrimps are widely distributed in coastal areas, estuaries and rivers. Although this shellfish is a good source of nutrients, it can also accumulate environmental contaminants, such as organochlorine pesticides (OCPs), brominated flame retardants (BFRs), synthetic musks (SMs) and polycyclic aromatic hydrocarbons (PAHs). Due to their bioaccumulative properties, these pollutants are endocrine disruptors. In this review, an overview of the world's shrimp market, pollutants legislation and values found in shrimp samples will be discussed. Shrimps analysed from all continents showed the presence of contaminants, Asia being the continent with the highest values reported. The concentration values reached a maximum of 26100 ng/g wet weight (ww) for OCPs, of 226.45 ng/g ww for BFRs, of 12.1 ng/g ww for SMs and of 50650 ng/g ww for PAHs. Exposure data and risk, taken from different studies, are very variable and indicate that shrimp's consumption may represent a risk especially in certain geographic areas., (© 2020 Published by Elsevier Ltd.)

One of the main challenges for analytical laboratories and food safety authorities is the control of food contaminants hazardous to human health. For the first time, a simple, fast, and cost-effective sample preparation method is proposed as an extraction technique to determine 12 brominated flame retardants (BFRs) (seven polybrominated diphenyl ether (PBDE) congeners and five novel BFRs) in Capsicum cultivars. Different QuEChERS and dispersive solid-phase extract (d-SPE) sorbent compositions were evaluated in terms of recovery and matrix effects. The best results were obtained with citrate-buffered version QuEChERS and a cleanup step, with 150 mg of MgSO 4 , 50 mg of primary secondary amine (PSA), 50 mg of C18, and 5 mg of carbon. The limit of detection (LOD) was between 1.4 and 9.3 μg/kg and R 2 > 0.99. Recoveries and matrix effects were between 66 and 104% and 0.58 and 2.18, respectively. The relative standard deviations from repeatability and reproducibility studies and estimation of measurement uncertainty were lower than 20%. Gas chromatography coupled with a mass spectrometer was used to confirm the presence of BFRs in the samples. Novel BFRs were detected lower than the LOD.

This study aimed to develop new canned chub mackerel products incorporating edible seaweeds ( A scophyllum nodosum , F ucus spiralis , S accorhiza polyschides , C hondrus crispus , Porphyra sp. and Ulva sp.) harvested in the Portuguese North-Central coast, with simultaneous sensory improvement and minerals enrichment. Two processes were compared, namely the addition of seaweeds in i) the canning step and ii) in the brining step (as the replacement for salt). The concentrations of four macrominerals (Na, K, Ca and Mg), chloride, and twelve trace elements (Co, Cu, Fe, I, Li, Mn, Mo, Rb, Se, Sr, V and Zn) were determined by high-resolution continuum source flame atomic absorption spectrometry (HR-CS-FAAS) and inductively coupled plasma mass spectrometry (ICP-MS), respectively. Results showed that canned chub mackerel incorporating C. crispus and F. spiralis was found to be the preferred sensory option, also exhibiting contents enriched with Cl, Co, Cu, Fe, I, Li, Mg, Mn, Mo, Na, Rb, Se, and Sr. This effect was more pronounced when both seaweed species were added to replace the salt added in the brining step., Competing Interests: The authors declare no conflict of interest.

Synthetic pyrethroids (SPs) are one of the most common pesticides used worldwide. Their use has greatly increased in the last decades and its' continuous application lead to added pesticides concentration in soil. Consequently, SPs may enter the food chain, affecting the environment and human health. The degradation over time of the pyrethroid pesticide deltamethrin applied to cabbages was monitored. The evolution was followed both on cabbages and the surrounding soils, and the soil microbial community characterized by next-generation sequencing of the 16S rRNA gene. The main shift in the microbial community structure was observed during the first 30 days after pesticides' application. The modification in the microbial community composition, where an increased abundance of Nocardioides sp. and Sphingomonas sp. were observed, was correlated respectively with the conversions of deltamethrin and its metabolite, 3-phenoxybenzoic acid (3-PBA). Although deltamethrin was not found in any of the tested samples (soil and cabbage) after 180 days, it caused an environmental impact much further than the 7 days security interval. These findings suggest that deltamethrin application can disturb soil microbial community and that natural biodegradation can have an important part in pesticides soil decontamination., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Small, single-celled planktonic cyanobacteria are ubiquitous in the world's oceans yet tend not to be perceived as secondary metabolite-rich organisms. Here we report the isolation and structure elucidation of hierridin C, a minor metabolite obtained from the cultured picocyanobacterium Cyanobium sp. LEGE 06113. We describe a simple, straightforward synthetic route to the scarcely produced hierridins that relies on a key regioselective halogenation step. In addition, we show that these compounds originate from a type III PKS pathway and that similar biosynthetic gene clusters are found in a variety of bacterial genomes, most notably those of the globally distributed picocyanobacteria genera Prochlorococcus, Cyanobium and Synechococcus.

Persistent organic pollutants (POPs) are man-made compounds with metabolic disruption impact. We investigated the effect of POP exposure in the cardiometabolic and inflammatory profile in a population of women with obesity and hypertension. In 43 premenopausal women (22 treated vs. 21 nontreated) undergoing bariatric surgery, blood and adipose tissue samples (visceral (vAT) and abdominal subcutaneous (scAT)) were collected. Median concentrations of ∑HCH and ∑POPs in vAT were significantly higher in treated women. VAT ∑HCH and scAT ∑HCH and ∑POPs concentrations were positively correlated with systolic blood pressure in the non-treated group. Our findings suggest that exposure to POPs and its accumulation in vAT and circulating in plasma may be associated to a higher cardiovascular risk in women with obesity and hypertension, with or without antihypertensive treatment.

3-Phenoxybenzoic acid (3-PBA) is a shared metabolite of several synthetic pyrethroid pesticides (SPs) resulting from environmental degradation of parent compounds and thus occurs frequently as a residue in samples. Hence, the importance of 3-PBA evaluation after pyrethroid application. There is a gap of analytical methods to determine 3-PBA in soil samples. Therefore, an analytical method that combines the solid-phase extraction (SPE) and gas chromatography-mass spectrometry (GC/MS) detection has been developed for the determination of 3-PBA in soil samples. The analytical method was validated in terms of linearity, sensitivity, intra- and inter-day batch precisions, recoveries, and quantification limits. An SPE method using a Strata X cartridge allows obtaining limits of detection and quantification equal to 4.0 and 13.3 ng g -1 , respectively. Under optimized conditions, the method average recovery levels ranged from 70.3 to 93.5% with a relative standard deviation below 3.4%. Method intra- and inter-day precision was under 5.0 and 4.8%, respectively. The developed method was applied to 11 agricultural soil samples in the north of Portugal. The developed methodology allowed for the determination of the pyrethroid metabolite, 3-PBA, in agricultural soil samples at levels of few ng g -1 . Graphical abstract ᅟ.

The total protein content and the (total and free) amino acid composition of nine edible species of red, brown and green seaweeds collected in the Portuguese North-Central coast were quantified to assess their potential contribution to the recommended dietary intake. Whenever possible, the protein and amino acid composition was compared with that of commercial European seaweeds. The protein content was the highest (P < 0.05) in red species (19.1-28.2 g/100 g dw), followed by the green seaweed Ulva spp. (20.5-23.3 g/100 g dw), with the lowest content found in brown seaweeds (6.90-19.5 g/100 g dw). Brown seaweeds presented the lowest mean contents of essential amino acids (EAAs) (41.0% protein) but significantly (P < 0.05) higher concentrations of non-essential amino acids (36.1% protein) and free amino acids (6.47-24.0% protein). Tryptophan, methionine and leucine were the limiting EAAs in all species. In contrast, lysine was found in high concentrations, especially in red (2.71-3.85% protein) and green (2.84-4.24% protein) seaweeds., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Magnetic nanoparticles (MNPs) with different sizes and characteristics were synthesized to be used as a QuEChERS sorbents for the determination of seven organophosphorus pesticides (OPPs) in strawberries by gas chromatography analysis with flame photometric and mass spectrometry detection. To achieve the optimum conditions of modified QuEChERS procedure several parameters affecting the cleanup efficiency including the amount of the sorbents and cleanup time were investigated. The results were compared with classical QuEChERS methodologies and the modified QuEChERS procedure using MNPs showed the better performance. Under the optimum conditions of the new methodology, three spiking levels (25, 50 and 100 μg kg -1 ) were evaluated in a strawberry sample. The results showed that the average recovery was 93% and the relative standard deviation was less than 12%. The enrichment factor ranged from 111 to 145%. The good linearity with coefficients of determination of 0.9904-0.9991 was obtained over the range of 25-250 μg kg -1 for 7 OPPs. It was determined that the MNPs have an excellent function as sorbent when purified even using less amount of sorbents and the magnetic properties allowed non-use of the centrifugation in cleanup step. The new methodology was applied in strawberry samples from conventional and organic farming. The new sorbents were successfully applied for extraction and determination of OPPs in strawberries., (Copyright © 2018 Elsevier B.V. All rights reserved.)

In this study we analyzed one of the most comprehensive sets of 21 urinary phthalate metabolites representing exposure to 11 parent phthalates (DEP, DMP, DiBP, DnBP, BBzP, DEHP, DiNP, DiDP, DCHP, DnPeP, DnOP) in first morning urine samples of 112 Portuguese children (4-18 years) sampled in 2014/15. The study population consisted of two groups: group 1 with normal weight/underweight children (N = 43) following their regular diet and group 2 with obese/overweight children (N = 69) following a healthy diet (with nutritional counselling). Most of the metabolites were above the limits quantification (81-100%) except for MCHP, MnPEP and MnOP. Metabolite levels were generally comparable to other recent child and general populations sampled worldwide, confirming the steady decline in exposures to most phthalates. Compared to Portuguese children sampled in 2011/2012, median urinary metabolite levels decreased by approximately 50% for DEHP, DnBP, DiBP and BBzP. Risk assessments for individual phthalates and the sum of the anti-androgenic phthalates did not indicate to attributable health risks, also at the upper percentiles of exposure. In the healthy diet group the median concentration of the DEHP metabolites was significant lower, while all phthalate metabolites except MEP tended to be lower compared to the regular diet group. Multiple log-linear regression analyses revealed significantly lower daily intakes (DIs) for all phthalates in the healthy diet group compared to the regular diet group (geometric mean ratios (gMR) between 0.510-0.618; p ≤ 0.05), except for DEP (gMR: 0.811; p = 0.273). The same analyses with the continuous variable body mass index instead of the diet groups also showed effects on the DIs (gMRs between 0.926-0.951; p ≤ 0.05), however much smaller than the effects of the diet. The results indicate that obese children following a healthy diet composed of fresh and less packaged/processed food can considerably reduce their intake for most phthalates and can have lower phthalate intakes than regular weight/regular diet children., (Copyright © 2018 Elsevier GmbH. All rights reserved.)

Pyrethroid pesticides residues have been frequently detected in soils and have been recognized to contribute to soil toxicity. The phytotoxic impact of pesticides was evaluated in Cucumis sativus (C. sativus) seeds. Percentage of seed germination, root elongation, shoot length and leaf length were considered as endpoints to assess the possible acute phytotoxicity of soil by the exposure to pyrethroid pesticides (cypermethrin, deltamethrin and cyhalothrin) and its metabolite phenoxybenzoic acid (3-PBA), in a concentration range between 50 and 500μgkg -1 . For germination percentage, it was only observed a negative impact when seeds were exposed to the metabolite. Cypermethrin showed impact in the three studied endpoints of seed development, while deltamethrin merely affected the root length. Concerning pigments content, it can be said that chlorophylls and total carotenoids median values increased for cypermethrin and deltamethrin. This increase was more pronounced to deltamethrin in joint effect with the organic solvent dimethyl sulphoxide (DMSO). When exposed to cyhalothrin and 3-PBA, no statistically significant differences were observed for C. sativus seeds to all the assessed endpoints of seed development and the investigated pigments content. This research brings new data concerning the relative sensitivity of C. sativus seeds to pyrethroids pesticides commonly found in agricultural facilities, as well as critical understanding and development of using C. sativus for phytotoxicity assessments efforts for pesticide exposures., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Bisphenol A (BPA) is considered an endocrine disruptor and public concern over BPA exposure has been raised. Several studies have assessed human exposure to this plasticizer, confirming its ubiquitous presence and highlighting children as a public of special concern. A simple, efficient, cheap and green analytical procedure is reported within this paper. This paper reports, for the first time, the development of a modified Micro-QuEChERS (Quick, Easy, Cheap, Effective, Rugged and Safe) method coupled to gas chromatography-mass spectrometry (GC-MS) as a new strategy for the efficient extraction and determination of Bisphenol A in human urine samples. Several parameters that are known to influence extraction were optimized. Good linearity was achieved at the studied concentration range (1-50μg/L), with a correlation coefficient (R 2 ) of 0.998. The optimized method proved to be accurate (≥74% recovery), reproducible (<11% relative standard deviation) and sensitive for BPA determination (detection limit of 0.13μg/L and quantification limit of 0.43μg/L). The analytical procedure was applied to the analyses of 12 urine samples collected from children living in the North/Center region of Portugal. BPA was detected in all the analyzed samples in concentrations ranging from 1.5μg/L to 48.9μg/L. The proposed methodology is suitable for the determination of BPA in urine samples in the framework of biomonitoring studies and bioanalytical analyses, applying GC-MS detection., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Exposure to bisphenol A (BPA) is known to be widespread and available data suggests that BPA can act as an endocrine disruptor. Diet is generally regarded as the dominant BPA exposure source, namely through leaching to food from packaging materials. The aim of this study was to evaluate the exposure of 110 Portuguese children (4-18 years old), divided in two groups: the regular diet group (n = 43) comprised healthy normal weight/underweight children with no dietary control; the healthy diet group (n = 67) comprised children diagnosed for obesity/overweight (without other known associated diseases) that were set on a healthy diet for weight control. First morning urine samples were collected and total urinary BPA was analyzed after enzymatic hydrolysis via on-line HPLC-MS/MS with isotope dilution quantification. Virtually, all the children were exposed to BPA, with 91% of the samples above the LOQ (limit of quantification) of 0.1 μg/L. The median (95th percentile) urinary BPA levels for non-normalized and creatinine-corrected values were 1.89 μg/L (16.0) and 1.92 μg/g creatinine (14.4), respectively. BPA levels in the regular diet group were higher than in the healthy diet group, but differences were not significant. Calculated daily BPA intakes, however, were significantly higher in children of the regular diet group than in children of healthy diet group. Median (95th percentile) daily intakes amounted to 41.6 (467) ng/kg body weight/day in the regular diet group, and 23.2 (197) ng/kg body weight/day in the healthy diet group. Multiple logistic regression analysis revealed that children in the healthy diet group had 33% lower intakes than children in the regular diet group (OR 0.67; 95% CI 0.51-0.89). For both groups, however, urinary BPA levels and daily BPA intakes were within the range reported for other children's populations and were well below health guidance values such as the European Food Safety Authority (EFSA) temporary tolerable daily intake (t-TDI) of 4 μg/kg body weight/day. In addition, lower daily BPA intakes were more likely linked with the inherent dietary approach rather than with high BMI or obesity.

Classical ortho-phthalate plasticizers are, due to their endocrine disrupting potency and reproductive toxicity, increasingly replaced by alternative plasticizers. Di(2-ethylhexyl) terephthalate (DEHTP) is one of these substitutes. In this study, we investigated DEHTP exposure in 107 Portuguese children (4-17years old) by analyzing specific DEHTP metabolites in their urine using a newly developed LC-MS/MS method. We could detect the major, specific DEHTP metabolite mono(2-ethyl-5-carboxypentyl) terephthalate (5cx-MEPTP) in 100% of the samples with levels above the limit of quantification in 96% of the samples (median concentration 4.19μg/L; 95th percentile 26.4μg/L; maximum 3400μg/L). Other minor DEHTP metabolites (5OH-MEHTP, 5oxo-MEHTP and 2cx-MMHTP) were detected at lower rates and levels. Daily DEHTP intakes calculated from urinary 5cx-MEPTP levels were generally far below the tolerable daily intake (TDI) of 1000μg/kgbw/d (median 0.67μg/kgbw/d; 95th percentile 6.25μg/kgbw/d; maximum 690μg/kgbw/d). However, for one child the biomarker-derived health-based guidance value (HBM-I value) for 5cx-MEPTP of 1800μg/L was exceeded by about a factor of two. Levels of 5cx-MEPTP and calculated daily DEHTP intakes were higher in normal/under-weight children who nourished on their usual diet compared to overweight/obese children who received nutritional guidance with fresh and unprocessed food (p=0.043 and p<0.001 respectively). This indicates to processed and fatty foodstuff as a major source of DEHTP exposure. Additionally, we found children of lower age having higher DEHTP intakes (p=0.045). Again, foodstuff as a major DEHTP source, together with other child specific DEHTP sources such as mouthing of toys or ingestion of dust might be contributing factors. With the present study, we provide a first data set on the omnipresent DEHTP exposure in children. So far, general levels of DEHTP exposure seem no cause for concern. However, due to the increasing use of DEHTP as an ortho-phthalate substitute, possible increasing exposures in the future should be followed closely., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Endocrine-disrupting chemicals such as p,p'-dichlorodiphenyldichloroethylene (p,p'-DDE), are bioaccumulated in the adipose tissue (AT) and have been implicated in the obesity and diabetes epidemic. Thus, it is hypothesized that p,p'-DDE exposure could aggravate the harm of an obesogenic context. We explored the effects of 12 weeks exposure in male Wistar rats' metabolism and AT biology, assessing a range of metabolic, biochemical and histological parameters. p,p'-DDE -treatment exacerbated several of the metabolic syndrome-accompanying features induced by high-fat diet (HF), such as dyslipidaemia, glucose intolerance and hypertension. A transcriptome analysis comparing mesenteric visceral AT (vAT) of HF and HF/DDE groups revealed a decrease in expression of nervous system and tissue development-related genes, with special relevance for the neuropeptide galanin that also revealed DNA methylation changes at its promoter region. Additionally, we observed an increase in transcription of dipeptidylpeptidase 4, as well as a plasmatic increase of the pro-inflammatory cytokine IL-1β. Our results suggest that p,p'-DDE impairs vAT normal function and effectively decreases the dynamic response to energy surplus. We conclude that p,p'-DDE does not merely accumulate in fat, but may contribute significantly to the development of metabolic dysfunction and inflammation. Our findings reinforce their recognition as metabolism disrupting chemicals, even in non-obesogenic contexts.

Escherichia coli is one of the most common etiological agents of diarrhea in developing countries. The appearance of resistant E. coli prevents treatment of these infections. Biotechnological products incorporating antimicrobial peptides are currently being considered in applications to prevent intestinal infections by these bacteria. The aim of this study was to evaluate the antibacterial activity of the peptide PcL342-354C, which is derived from the toxin Cry1Ab16 from Bacillus thuringiensis, against E. coli strains. We also report the preparation, characterization and evaluation of the antibacterial activity of LbL films containing PcL342-354C. The results showed that the PcL342-354C peptide inhibited the growth of different strains of E. coli with minimal inhibitory concentration ranging from 15.62-31.25μg/mL and minimal bactericidal concentration was 250μg/mL, indicating a potential antibacterial activity. The morphology of an ITO/Cashew gum/PcL342-354C film was analysed using atomic force microscopy which showed an increase of roughness due to the increase in the number of layers. The LbL films showed significant antibacterial activity against E. coli NCTC 9001 in both conditions tested (10 and 20 bilayers). Our results indicate that the peptide exhibits an antibacterial potential that can be tapped to develop biomaterials with antibacterial activity for use against foodborne pathogens., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Di-(iso-nonyl)-cyclohexane-1,2-dicarboxylate (DINCH) is used as substitute for high molecular weight phthalate plasticizers such as di-(2-ethylhexyl) phthalate (DEHP) and di-(iso-nonyl) phthalate (DINP). Due to a rapid substitution process we have to assume omnipresent and increasing DINCH exposures. The aim of this study was to evaluate DINCH exposure in 112 children (4-18years old) from Portugal, divided in two groups: 1) normal-/underweight following the usual diet; and 2) obese/overweight but under strict nutritional guidance. First morning urine samples were collected during the years 2014 and 2015. Oxidized DINCH metabolites (OH-MINCH, oxo-MINCH, cx-MINCH) were analyzed after enzymatic hydrolysis via on-line HPLC-MS/MS with isotope dilution quantification. We detected DINCH metabolites in all analyzed samples. Urinary median (95th percentile) concentrations were 2.14μg/L (15.91) for OH-MINCH, followed by 1.10μg/L (7.54) for oxo-MINCH and 1.08μg/L (7.33) for cx-MINCH. We observed no significant differences between the two child-groups; only after creatinine adjustment, we found higher metabolite concentrations in the younger compared to the older children. Median (95th percentile) daily DINCH intakes were in the range of 0.37 to 0.76 (2.52 to 5.61) μg/kg body weight/day depending on calculation model and subpopulation. Body weight related daily intakes were somewhat higher in Group 1 compared to Group 2, irrespective of the calculation model. However, in terms of absolute amounts (μg/day), DINCH intakes were higher in Group 2 compared to Group 1. In regard to age, we calculated higher intakes for the younger children compared to older children, but only with the creatinine-based model. This new data for southern European, Portuguese children adds information to the scarce knowledge on DINCH, confirming omnipresent exposure and suggesting higher exposures in children than adults. Significant sources and routes of exposure have yet to be unveiled. For now, all calculated daily intakes are far below established health benchmark levels (TDI, RfD). However, rapidly increasing exposures have to be expected over the next years., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

The present work was designed to study the mechanisms associated with Nurr1 modulation following early life permethrin (PERM) treatment during rat's life span. Here we demonstrate that PERM exposure in rats, at a dose close to No Observed Adverse Effect Level (NOAEL) for 15days during neonatal brain development leads to its accumulation long after exposure. In striatum from adolescent rats we detected an increase in DNA methyltransferases (DNMTs) such as DNMT1, DNMT3a, Tyrosine hydroxylase, monomeric and aggregated α-synuclein protein levels. Adult rats showed enhanced DNMT3b and α-synuclein aggregation compared to the control group, while with aging a significant decrease in all biomarkers studied was observed. No changes in Nurr1 promoter methylation in adolescent, adult and old rats were found. In silico studies showed clear evidence of a strong binding interaction between PERM and its metabolite 3-phenoxybenzoic acid with the nuclear orphan receptor Nurr1. These findings suggest that an additional interference with the dopaminergic neuron pathway could occur in situ during PERM accumulation in brain. Therefore, Nurr1 modulation in early life PERM-treated rats, depends on age-related adaptive responses in animals., (Copyright © 2016 IBRO. Published by Elsevier Ltd. All rights reserved.)

The Quick, Easy, Cheap, Effective, Rugged, and Safe (QuEChERS) method was applied to the extraction of 14 organochlorine pesticides (OCPs) residues from commercial fruit pulps available in supermarkets in Fortaleza, Northeastern Brazil. The analyses were carried out by gas chromatography (GC), coupled to an electron-capture detector (ECD), and were confirmed by GC-tandem mass spectrometry (MS). The parameters of the analytical method, such as accuracy, precision, linear range, limits of detection and quantification, were determined for each pesticide. The results showed good linearity (R 2 ≥ 0.9916) and the overall average recoveries were considered satisfactory obtaining values between 69 and 110%, RSD of 2-15 %, except for hexachlorobenzene (HCB) in açai, acerola and guava pulp samples. The OCPs were detected in guava (α-HCH; lindane) and soursop (α, β-HCH isomers) samples. The QuEChERS method and GC-ECD were successfully used to analyze OCPs in commercially available Brazilian fruit pulps and can be applied in routine analytical laboratories.

Context: Several studies have shown a link between proinflammatory activity and the presence or deficit of some fatty acids. Inflammation is associated with several diseases including diabetes., Objective: To characterize and compare the fatty acids profiles in children with inaugural type 1 diabetes, diabetic children (at least 1 year after diagnosis), and healthy children., Design: Plasma fatty acids profiles in children with inaugural diabetes, children with noninaugural diabetes, and controls, all of whom were prepubescent with a BMI < 85th percentile, were evaluated., Results: Omega-3 fatty acid levels were higher in recently diagnosed subjects with diabetes than in controls. The ratio of omega-6/omega-3 fatty acids was higher in the control population. Omega-6 fatty acid levels were higher in the nonrecent diabetic subjects than in the children with recently diagnosed diabetes, and the levels were higher in the nonrecent diabetes group compared to the control group., Conclusion: Our findings showed higher levels of alpha-linolenic acid, EPA, and DHA, as well as mono- and polyunsaturated fatty acids, in diabetic children. These findings reinforce the importance of precocious nutritional attention and intervention in the treatment of diabetic children.

The number of children affected by Autism Spectrum Disorders (ASD) is dramatically increasing as well as the studies aimed at understanding the risk factors associated with the development of ASD. Since the etiology of ASD is partly genetic and partly environmental, factors (i.e., heavy metals, pesticides) as well as lifestyle seem to have a key role in the development of the disease. ASD and Control (CTR) children, aged 5-12 years, were compared. Gas chromatography coupled with trap mass detector was used to measure the level of 3-PBA, the main pyrethroid metabolite in urine in a group of ASD patients, while optical emission spectrometry analysis was employed to estimate the level of metals and microelements in hair in a different group of ASD children. The presence of 3-PBA in urine seems to be independent of age in ASD children, while a positive correlation between 3-PBA and age was observed in the control group of the same age range. Urine concentration of 3-BPA in ASD children had higher values than in the control group, which were marginally significant (p = 0.054). Mg results were significantly decreased in ASD with respect to controls, while V, S, Zn, and Ca/Mg were marginally increased, without reaching statistical significance. Results of Principal Component (PC) analysis of metals and microelements in hair were not associated with either age or health status. In conclusion, 3-PBA in urine and Mg in hair were changed in ASD children relative to control ones.

Background: Erectile dysfunction (ED) is a prevalent complication of diabetes, and oxidative stress is an important feature of diabetic ED. Oxidative stress-induced damage plays a pivotal role in the development of tissue alterations. However, the deleterious effects of oxidative stress in the corpus cavernosum with the progression of diabetes remain unclear. The aim of this study was to evaluate systemic and penile oxidative stress status in the early and late stages of diabetes., Methods: Male Wistar streptozotocin-diabetic rats (and age-matched controls) were examined 2 (early) and 8 weeks (late) after the induction of diabetes. Systemic oxidative stress was evaluated by urinary H2 O2 and the ratio of circulating reduced/oxidized glutathione (GSH/GSSG). Penile oxidative status was assessed by H2 O2 production and 3-nitrotyrosine (3-NT) formation. Cavernosal endothelial nitric oxide synthase (eNOS) was analyzed by quantitative immunohistochemistry. Dual immunofluorescence was also performed for 3-NT and α-smooth muscle actin (α-SMA) and eNOS-α-SMA., Results: There was a significant increase in urinary H2 O2 levels in both diabetic groups. The plasma GSH/GSSG ratio was significantly augmented in late diabetes. In cavernosal tissue, H2 O2 production was significantly increased in late diabetes. Reactivity for 3-NT was located predominantly in cavernosal smooth muscle (SM) and was significantly reduced in late diabetes. Quantitative immunohistochemistry revealed a significant decrease in eNOS levels in cavernosal SM and endothelium in late diabetes., Conclusions: The findings indicate that the noxious effects of oxidative stress are more prominent in late diabetes. Increased penile protein oxidative modifications and decreased eNOS expression may be responsible for structural and/or functional deregulation, contributing to the progression of diabetes-associated ED., (© 2014 Ruijin Hospital, Shanghai Jiaotong University School of Medicine and Wiley Publishing Asia Pty Ltd.)

Eight tropical fruit pulps from Brazil were simultaneously characterised in terms of their antioxidant and antimicrobial properties. Antioxidant activity was screened by DPPH radical scavenging activity (126-3987 mg TE/100g DW) and ferric reduction activity power (368-20819 mg AAE/100g DW), and complemented with total phenolic content (329-12466 mg GAE/100g DW) and total flavonoid content measurements (46-672 mg EE /100g DW), whereas antimicrobial activity was tested against the most frequently found food pathogens. Acerola and açaí presented the highest values for the antioxidant-related measurements. Direct correlations between these measurements could be observed for some of the fruits. Tamarind exhibited the broadest antimicrobial potential, having revealed growth inhibition of Pseudomonas aeruginosa. Escherichia coli, Listeria monocytogenes, Salmonella sp. and Staphylococcus aureus. Açaí and tamarind extracts presented an inverse relationship between antibacterial and antioxidant activities, and therefore, the antibacterial activity cannot be attributed (only) to phenolic compounds., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Eruca sativa (rocket salad) has been intensely consumed all over the world, insomuch as, this work was undertaken to evaluate the antioxidant status and the environmental contamination (positive and negative nutritional contribution) of leaves and stems from this vegetable. Antioxidant capacity of rocket salad was assessed by mean of optical methods, such as the total phenolic content (TPC), reducing power assay and DPPH radical scavenging activity. The extent of the environmental contamination was reached through the quantification of thirteen organochlorine pesticides (OCP) by using gas chromatography coupled with electron-capture detector (GC-ECD) and compound confirmations employing gas chromatography tandem mass-spectrometry (GC-MS/MS). The OCP residues were extracted by using Quick, Easy, Cheap, Effective, Rugged and Safe (QuEChERS) methodology.The extent of the environmental contamination was reached through the quantification of thirteen OCP by using gas chromatography coupled with electron-capture detector (GC-ECD) and compound confirmations employing GC-MS/MS. The OCP residues were extracted by using Quick, Easy, Cheap, Effective, Rugged and Safe (QuEChERS) methodology. This demonstrated that leaves presented more antioxidant activity than stems, emphasizing that leaves contained six times more polyphenolic compounds than stems. In what concerns the OCP occurrence, the average recoveries obtained at the three levels tested (40, 60 and 80 µg kg(-1)) ranged from 55% to 149% with a relative standard deviation of 11%, (except hexachrorobenzene). Three vegetables samples were collected from supermarkets and analysed following this study. According to data, only one sample achieved 16.21 of β-hexachlorocyclohexane, confirmed by GC-MS/MS. About OCP quantification, the data indicated that only one sample achieved 16.21 µg kg(-1) of β-hexachlorocyclohexane, confirmed by GC-MS/MS, being the QuEChERS a good choice for the of OCPs extraction. Furthermore, the leaves consumption guaranty higher levels of antioxidants than stems.

In this study, we sought to assess the applicability of GC-MS/MS for the identification and quantification of 36 pesticides in strawberry from integrated pest management (IPM) and organic farming (OF). Citrate versions of QuEChERS (quick, easy, cheap, effective, rugged and safe) using dispersive solid-phase extraction (d-SPE) and disposable pipette extraction (DPX) for cleanup were compared for pesticide extraction. For cleanup, a combination of MgSO4, primary secondary amine and C18 was used for both the versions. Significant differences were observed in recovery results between the two sample preparation versions (DPX and d-SPE). Overall, 86% of the pesticides achieved recoveries (three spiking levels 10, 50 and 200 µg/kg) in the range of 70-120%, with <13% RSD. The matrix effects were also evaluated in both the versions and in strawberries from different crop types. Although not evidencing significant differences between the two methodologies were observed, however, the DPX cleanup proved to be a faster technique and easy to execute. The results indicate that QuEChERS with d-SPE and DPX and GC-MS/MS analysis achieved reliable quantification and identification of 36 pesticide residues in strawberries from OF and IPM., (© The Author [2014]. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)