Adipocyte proteome and secretome influence inflammatory and hormone pathways in glioma.

Gliomas represent the most common primary malignant brain tumors in adults, with an extremely poor prognosis. Among several risk factors, lifestyle was also recently identified as a major risk factor for the development of primary glioma. In the present study, we explore the relationship between obesity and glioma in a cellular model. Thus, we have study the influence of adipocytes secretome on glioma cell line GL261. Using the 3T3-L1 adipocyte cell line, and its conditioned medium (adipokines-enriched medium), we showed that adipocyte-released factors relate with glioma angiogenic, growth, hormones and metabolic behavior by MALDI-TOF-MS and proteomic array analysis. In a first view, STI1, hnRNPs and PGK1 are under expressed on CGl. Similarly, both carbonic anhydrase and aldose reductase are even suppressed in glioma cells that grown under adipokines-enriched environment. Contrariwise, RFC1, KIF5C, ANXA2, N-RAP and RACK1 are overexpressed in GL261 cell the in the presence of the adipokines-enriched medium. We further identified the factors that are released by adipocyte cells, and revealed that several pro-inflammatory and angiogenic factors, such as IL-6, IL-11, LIF, PAI-1, TNF-α, endocan, HGF, VEGF IGF-I, were secreted to the medium into a high extent, whereas TIMP-1 and SerpinE1 were under expressed on CGl. This study discloses an interesting in vitro model for the study of glioma biology under a "obesity" environment, that can be explored for the understanding of cancer cells biology, for the search of biomarkers, prognostic markers and therapeutic approaches.

17α-Ethinylestradiol and 17ß-estradiol removal from a secondary urban wastewater using an RBC treatment system.

The presence of micropollutants that include endocrine-disrupting compounds (EDC) in aquatic environments is currently one of the most relevant aspects of water quality due to their adverse effects on aquatic organisms and human health. From the several categories of EDC, 17ß-estradiol (E2) is a natural hormone, which is prevalent in vertebrates, associated with the female reproductive system and maintenance of the sexual characters. 17α-Ethinylestradiol (EE2) is a synthetic hormone produced from the natural hormone E2 and is an essential component of oral contraceptives. These compounds are susceptible to bioconcentration and have high potential to bioaccumulation. Wastewater treatment plants are the main point source of E2 and EE2 into aquatic environments, but conventional wastewater treatment systems are not specifically designed for steroid removal. To overcome this problem, biological tertiary treatment may be a solution for the removal of emergent pollutants such as E2 and EE2. The main purpose of the present study is to provide a solution based on the optimization of a rotating biological contactor system to remove estrogens, specifically E2 and EE2, and to quantify their removal efficiency on different matrices, namely real wastewater and different synthetic wastewaters. All assays presented viable removal efficiencies for compound E2 with values always above 50%; real wastewater yielded the highest removal efficiencies. EE2 removal had better removal efficiencies with synthetic wastewater as feed solution, with removals above 15%, whereas the removal efficiency with real wastewater was inexistent.

Screening of priority pesticides in Ulva sp. seaweeds by selective pressurized solvent extraction before gas chromatography with electron capture detector analysis.

This work reports a fast and reliable analytical method for the screening of priority pesticides (PPs) in Ulva sp. seaweeds by gas chromatography with electron capture detection. Extraction and sample clean-up were performed in one single step by selective pressurized liquid extraction (SPLE). Several parameters affecting SPLE performance were optimized. Method performance was compared with standard Soxhlet extraction. Significant decrease of the time of analysis with better recoveries for a greater number of PPs was achieved by SPLE. Average recoveries ranged from 71 to 103% with RSD < 10%. Field application showed the presence of PP in the range of 3-11 ng g(-1) in seaweeds collected in a coastal lagoon after a long period of heavy rains. These results suggest that Ulva sp. seaweeds tend to accumulate PPs and have the potential to be used as early alert signals of aquatic pollution especially after rains and storm events.

Analysis of 65 pharmaceuticals and personal care products in 5 wastewater treatment plants in Portugal using a simplified analytical methodology.

Pharmaceuticals and personal care products (PPCPs) are becoming increasingly recognised as important micropollutants to be monitored in wastewater treatment plants (WWTPs), since WWTP effluents represent an important point source to natural aquatic systems. In this study, the abundance of 65 PPCPs was analysed in 5 Portuguese WWTPs during the spring and autumn. Due to the fact that analytical approaches normally used to quantify the abundance of these compounds are labour intensive and require various specific procedures, this study proposes a set of simplified analytical methods for the quantification of pharmaceutically active compounds (PhACs) and polycyclic musks in liquid and sludge samples. The analytical methods were validated using influent wastewater matrices, showing comparable limits of detection and quantification as literature values for most PPCPs, with the exception of the estrogenic compounds. The PhAC concentrations detected in the WWTP survey were in the range of 0.050-100 µg L(-1) in the influent and up to 50 µg L(-1) in the effluent, where the non-steroidal anti-inflammatory drugs (NSAIDs) were the most abundant and frequently detected group. Some musks were detected up to 11.5 µg L(-1) in the influent and 0.9 µg L(-1) in the effluent, and adsorbed in the sludge up to 22.6 µg g(-1).

Metabolism and ecological niche of Tetrasphaera and Ca. Accumulibacter in enhanced biological phosphorus removal.

Tetrasphaera and Candidatus Accumulibacter are two abundant polyphosphate accumulating organisms in full-scale enhanced biological phosphorus removal (EBPR) systems. However, little is known about the metabolic behaviour and ecological niche that each organism exhibits in mixed communities. In this study, an enriched culture of Tetrasphaera and Ca. Accumulibacter was obtained using casein hydrolysate as sole carbon source. This culture was able to achieve a high phosphorus removal efficiency (>99%), storing polyphosphate while consuming amino acids anaerobically. Microautoradiography and fluorescence in situ hybridisation confirmed that more than 90% Tetrasphaera cells were responsible for amino acid consumption while Ca. Accumulibacter likely survived on fermentation products. Tetrasphaera performed the majority of the P removal (approximately 80%) in this culture, and batch tests showed that the metabolism of some carbon sources could actually lead to anaerobic orthophosphate (Pi) uptake (9.0 ± 2.1 mg-P/L) through energy generated by fermentation of glucose and amino acids. This anaerobic Pi uptake may lead to lower net Pi release to C uptake ratios and reduce the Pi needed to be removed aerobically in WWTPs. Intracellular metabolites such as amino acids, sugars, volatile fatty acids and small amines were observed as potential storage products, which may serve as energy sources in the aerobic phase. Evidence of the urea cycle was found, which could be involved in reducing the intracellular nitrogen content. This study improves our understanding of how phosphorus is removed in EBPR systems and can enable novel process optimisation strategies.

Pathways of priority pesticides in sediments of coastal lagoons: The case study of Óbidos Lagoon, Portugal.

This study reports the concentrations of the priority pesticides (PPs) in 14 surface sediments and 21 layers of a sediment core from Óbidos Lagoon, a shallow Portuguese coastal lagoon. Results show that the PPs are confined to the upper part of the lagoon that receives most of the inputs from surface runoff of the surrounding agricultural fields and from small tributaries. Past and recent applied PPs were registered in sediments, aluminum normalized concentrations varying between 0.05×10(-7) and 6.85×10(-7). The PP risk assessment based on sediment quality guidelines like the "Probable Effect Level" (PEL) shows no biological effects in either sediments or aquatic organisms of Óbidos Lagoon, except for dieldrin, lindane, DDT, heptachlor epoxide and its parent compound heptachlor.

Priority pesticides in sediments of European coastal lagoons: A review.

This review summarizes the legislation applied to pesticides and highlights the physicochemical properties of the past and recently listed PPs under Water Framework Directive (WFD). It reports important information regarding the analysis, occurrence and long-term screening of PPs in sediments of European coastal lagoons. Among the entire list of PPs, those analyzed have been the organochloride insecticides, such as lindane, hexachlorobenzene and DDT. Although flood events pointed to the possible redistribution of pesticides, which may increase under climate change conditions, few studies are available concerning PPs screening in sediments of European coastal lagoons. Data is scarce not only in terms of the number of listed PPs that have been analyzed but also in terms of their long-term screening. This lack of data on PP concentrations is probably a consequence of the fact that less importance is given to the sediments contamination/quality.

Ecotoxicity of ketoprofen, diclofenac, atenolol and their photolysis byproducts in zebrafish (Danio rerio).

The occurrence of pharmaceutical compounds in wastewater treatment plants and surface waters has been detected worldwide, constituting a potential risk for aquatic ecosystems. Adult zebrafish, of both sexes, were exposed to three common pharmaceutical compounds (atenolol, ketoprofen and diclofenac) and their UV photolysis by-products over seven days. The results show that diclofenac was removed to concentrations

Propionate addition enhances the biodegradation of the xenobiotic herbicide propanil and its metabolite.

This study investigated ways of stimulating the biodegradation rates of the commonly applied herbicide, 3,4-dichloropropionanilide (propanil), and its metabolite, 3,4-dichloroaniline (DCA), as well as the growth rate of propanil- and DCA-degrading organisms in a mixed culture. Propionate, the other metabolite of propanil, stimulated the specific degradation rates of both propanil and DCA after a brief acclimation period. A metabolic model developed to characterise the metabolism of propanil and DCA biodegradation showed that the efficiency of oxidative phosphorylation (i.e. P/O ratio), which measures the metabolic efficiency, increased over time by 6- to 10-fold. This increase was accompanied by a 5- to 10-fold increase in the propanil and DCA biodegradation degradation rates. The biodegradation rates of the culture were unaffected when using an irrigation water matrix (Tejo river, Portugal), highlighting the utility of the culture for bioaugmentation purposes.

Photodegradation kinetics and transformation products of ketoprofen, diclofenac and atenolol in pure water and treated wastewater.

Pharmaceutical compounds such as ketoprofen, diclofenac and atenolol are frequently detected at relatively high concentrations in secondary effluents from wastewater treatment plants. Therefore, it is important to assess their transformation kinetics and intermediates in subsequent disinfection processes, such as direct ultraviolet (UV) irradiation. The photodegradation kinetics of these compounds using a medium pressure (MP) lamp was assessed in pure water, as well as in filtered and unfiltered treated wastewater. Ketoprofen had the highest time- and fluence-based rate constants in all experiments, whereas atenolol had the lowest values, which is consistent with the corresponding decadic molar absorption coefficient and quantum yield. The fluence-based rate constants of all compounds were evaluated in filtered and unfiltered wastewater matrices as well as in pure water. Furthermore, transformation products of ketoprofen, diclofenac and atenolol were identified and monitored throughout the irradiation experiments, and photodegradation pathways were proposed for each compound. This enabled the identification of persistent transformation products, which are potentially discharged from WWTP disinfection works employing UV photolysis.

Biodegradation of clofibric acid and identification of its metabolites.

Clofibric acid (CLF) is the pharmaceutically active metabolite of lipid regulators clofibrate, etofibrate and etofyllinclofibrate, and it is considered both environmentally persistent and refractory. This work studied the biotransformation of CLF in aerobic sequencing batch reactors (SBRs) with mixed microbial cultures, monitoring the efficiency of biotransformation of CLF and the production of metabolites. The maximum removal achieved was 51% biodegradation (initial CLF concentration=2 mg L(-1)), where adsorption and abiotic removal mechanisms were shown to be negligible, showing that CLF is indeed biodegradable. Tests showed that the observed CLF biodegradation was mainly carried out by heterotrophic bacteria. Three main metabolites were identified, including α-hydroxyisobutyric acid, lactic acid and 4-chlorophenol. The latter is known to exhibit higher toxicity than the parent compound, but it did not accumulate in the SBRs. α-Hydroxyisobutyric acid and lactic acid accumulated for a period, where nitrite accumulation may have been responsible for inhibiting their degradation. A metabolic pathway for the biodegradation of CLF is proposed in this study.

Assessing the removal of pharmaceuticals and personal care products in a full-scale activated sludge plant.

PURPOSE: This study aimed to investigate the removal mechanisms of pharmaceutical active compounds (PhACs) and musks in a wastewater treatment plant (WWTP). Biological removal and adsorption in the activated sludge tank as well as the effect of UV radiation used for disinfection purposes were considered when performing a mass balance on the WWTP throughout a 2-week sampling campaign. METHODS: Solid-phase extraction (SPE) was carried out to analyse the PhACs in the influent and effluent samples. Ultrasonic solvent extraction was used before SPE for PhACs analysis in sludge samples. PhAC extracts were analysed by LC-MS. Solid-phase microextraction of liquid and sludge samples was used for the analysis of musks, which were detected by GC-MS. The fluxes of the most abundant compounds (13 PhACs and 5 musks) out of 79 compounds studied were used to perform the mass balance on the WWTP. RESULTS: Results show that incomplete removal of diclofenac, the compound that was found in the highest abundance, was observed via biodegradation and adsorption, and that UV photolysis was the main removal mechanism for this compound. The effect of adsorption to the secondary sludge was often negligible for the PhACs, with the exceptions of diclofenac, etofenamate, hydroxyzine and indapamide. However, the musks showed a high level of adsorption to the sludge. UV radiation had an important role in reducing the concentration of some of the target compounds (e.g. diclofenac, ibuprofen, clorazepate, indapamide, enalapril and atenolol) not removed in the activated sludge tank. CONCLUSIONS: The main removal mechanism of PhACs and musks studied in the WWTP was most often biological (45%), followed by adsorption (33%) and by UV radiation (22%). In the majority of the cases, the WWTP achieved >75% removal of the most detected PhACs and musks, with the exception of diclofenac.

Assessing the diurnal variability of pharmaceutical and personal care products in a full-scale activated sludge plant.

An intensive sampling campaign has been carried out in a municipal wastewater treatment plant (WWTP) to assess the dynamics of the influent pharmaceutical active compounds (PhAC) and musks. The mass loadings of these compounds in wastewater influents displayed contrasting diurnal variations depending on the compound. The musks and some groups of PhACs tended to follow a similar diurnal trend as compared to macropollutants, while the majority of PhACs followed either the opposite trend or no repeatable trend. The total musk loading to the WWTP was 0.74 ± 0.25 g d(-1), whereas the total PhAC mass loading was 84.7 ± 63.8 g d(-1). Unlike the PhACs, the musks displayed a high repeatability from one sampling day to the next. The range of PhAC loadings in the influent to WWTPs can vary several orders of magnitude from one day or week to the next, representing a challenge in obtaining data for steady-state modelling purposes.

Flavored waters: influence of ingredients on antioxidant capacity and terpenoid profile by HS-SPME/GC-MS.

The antioxidant profiles of 39 water samples (29 flavored waters based on 10 natural waters) and 6 flavors used in their formulation (furnished by producers) were determined. Total phenol and flavonoid contents, reducing power, and DPPH radical scavenging activity were the optical techniques implemented and included in the referred profile. Flavor extracts were analyzed by HS-SPME/GC-MS to obtain the qualitative and quantitative profiles of the volatile fraction of essential oils. Results pointed out a higher reducing power (0.14-11.8 mg of gallic acid/L) and radical scavenging activity (0.29-211.5 mg Trolox/L) of flavored waters compared with the corresponding natural ones, an interesting fact concerning human health. Bioactive compounds, such as polyphenols, were present in all samples (0.5-359 mg of gallic acid/L), whereas flavonoids were not present either in flavored waters or in flavors. The major components of flavor extracts were monoterpenes, such as citral, α-limonene, carveol, and α-terpineol.

Microcystin-LR detection in water by the Fabry-Pérot interferometer using an optical fibre coated with a sol-gel imprinted sensing membrane.

Cyanobacteria deteriorate the water quality and are responsible for emerging outbreaks and epidemics causing harmful diseases in Humans and animals because of their toxins. Microcystin-LR (MCT) is one of the most relevant cyanotoxin, being the most widely studied hepatotoxin. For safety purposes, the World Health Organization recommends a maximum value of 1 µg L(-1) of MCT in drinking water. Therefore, there is a great demand for remote and real-time sensing techniques to detect and quantify MCT. In this work a Fabry-Pérot sensing probe based on an optical fibre tip coated with a MCT selective thin film is presented. The membranes were developed by imprinting MCT in a sol-gel matrix that was applied over the tip of the fibre by dip coating. The imprinting effect was obtained by curing the sol-gel membrane, prepared with (3-aminopropyl) trimethoxysilane (APTMS), diphenyl-dimethoxysilane (DPDMS), tetraethoxysilane (TEOS), in the presence of MCT. The imprinting effect was tested by preparing a similar membrane without template. In general, the fibre Fabry-Pérot with a Molecular Imprinted Polymer (MIP) sensor showed low thermal effect, thus avoiding the need of temperature control in field applications. It presented a linear response to MCT concentration within 0.3-1.4 µg L(-1) with a sensitivity of -12.4±0.7 nm L µg(-1). The corresponding Non-Imprinted Polymer (NIP) displayed linear behaviour for the same MCT concentration range, but with much less sensitivity, of -5.9±0.2 nm L µg(-1). The method shows excellent selectivity for MCT against other species co-existing with the analyte in environmental waters. It was successfully applied to the determination of MCT in contaminated samples. The main advantages of the proposed optical sensor include high sensitivity and specificity, low-cost, robustness, easy preparation and preservation.

Biological treatment of propanil and 3,4-dichloroaniline: kinetic and microbiological characterisation.

Propanil (3,4-dichloropropionanilide) is a widely used herbicide, applied worldwide in rice paddies. Propanil is primarily transformed in nature to 3,4-dichloroaniline (DCA), which is more slowly biodegradable. Both compounds have adverse health and ecotoxicity effects. This work investigated the microbial ecology and kinetics of propanil-degrading enrichments obtained from soil in a sequencing batch reactor (SBR) operated with different feeding strategies, aiming at the enhanced biological removal of propanil and DCA from contaminated waters. During SBR operation with a dump feeding strategy, a high propanil concentration led to DCA accumulation, which was only fully degraded after 5 days, likely due to DCA inhibition. For this reason, the operational mode was changed to fed-batch operation with lower initial propanil concentrations, which resulted in faster propanil and DCA biodegradation. Thus a fed-batch operation seems more appropriate for the acclimatisation of an effective propanil- and DCA-degrading population. The changes in performance were accompanied by a shift in the microbial population structure, as determined by DGGE of the 16S rRNA gene, particularly after a feed of DCA as the sole carbon source. Isolates obtained from the acclimatised population included members of the genera Enterococcus and Rhodococcus, as well as Brevundimonas, which displayed >90% propanil biodegradation efficiency.