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Background: Developmental Coordination Disorder (DCD) affects school-age children and interferes with the practice of their daily activities. Task-oriented work and motor imagery have shown great efficacy in addressing this problem., Objective: This study presents a protocol that seeks to understand the effectiveness of a combined program of both modalities on the motor competence of children susceptible to the presence of DCD., Desing: Randomised controlled trial., Methods: The participants are children susceptible to the presence of DCD (from 6 to 12 years old) distributed into an intervention group that received a total of 20 protocol sessions and a control group that continued with their usual school routine.

Introduction and Objective: Generative artificial intelligence makes it possible to ask about medical pathologies in dialog boxes. Our objective was to analyze the quality of information about the most common urological pathologies provided by ChatGPT (OpenIA), BARD (Google), and Copilot (Microsoft)., Methods: We analyzed information on the following pathologies and their treatments as provided by AI: prostate cancer, kidney cancer, bladder cancer, urinary lithiasis, and benign prostatic hypertrophy (BPH). Questions in English and Spanish were posed in dialog boxes; the answers were collected and analyzed with DISCERN questionnaires and the overall appropriateness of the response. Surgical procedures were performed with an informed consent questionnaire., Results: The responses from the three chatbots explained the pathology, detailed risk factors, and described treatments. The difference is that BARD and Copilot provide external information citations, which ChatGPT does not. The highest DISCERN scores, in absolute numbers, were obtained in Copilot; however, on the appropriacy scale it was noted that their responses were not the most appropriate. The best surgical treatment scores were obtained by BARD, followed by ChatGPT, and finally Copilot., Conclusions: The answers obtained from generative AI on urological diseases depended on the formulation of the question. The information provided had significant biases, depending on pathology, language, and above all, the dialog box consulted., (Copyright © 2024 AEU. Published by Elsevier España, S.L.U. All rights reserved.)

Curcumin is a natural phenolic compound with important biological functions. Despite its demonstrated efficacy in vitro, curcumin biological activities in vivo are dependent on its bioaccessibility and bioavailability, which have been highlighted as a crucial challenge. Cetyltrimethylammonium bromide-modified cellulose nanocrystals (CNC-CTAB) have been shown to be effective in curcumin encapsulation, as they have the potential to enhance biological outcomes. This study evaluated the biological effects of curcumin encapsulated within CNC-CTAB structures, namely its antioxidant, anti-inflammatory and antimicrobial properties, as well as the release profile under digestion conditions and intestinal permeability. Encapsulated curcumin demonstrated antioxidant and anti-inflammatory properties, effectively reducing reactive oxygen species and cytokine production by intestinal cells. The delivery system exhibited antimicrobial properties against Campylobacter jejuni bacteria, further suggesting its potential in mitigating intestinal inflammation. The system showed the ability to protect curcumin from degradation and facilitate its interaction with the intestinal epithelium, highlighting the potential of CNC-CTAB as carrier to enhance curcumin intestinal biological functions.

Introduction: The vacuum-assisted access sheath is a new device for the treatment of kidney stones with percutaneous nephrolithotomy (PCNL)., Objective: Our aim was to compare the stone-free rate (SFR) and complications between standard mini percutaneous nephrolithotomy (Mini-PCNL) and vacuum-assisted PCNL (Va-PCNL)., Methods: Retrospective study of patients undergoing Mini-PCNL and Va-PCNL from January 2018 to June 2022. Va-PCNL was performed with a disposable sheath (ClearPetra®) with continuous high-flow irrigation and vacuum fluid dynamics for easier stone fragment removal. Baseline patient characteristics, surgical outcomes, perioperative and postoperative data were collected. We compared SFR and complications., Results: A total of 136 patients were identified, 57 (41,9%) underwent Va-PCNL and 79 (58,15%) Mini-PCNL. Mean operative time was significantly shorter in the Va-PCNL group (95 min.) than in Mini-PCNL (146 min; P = ,001) group. The tubeless technique was performed more frequently in Va-PCNL group (61,4% vs. 34,2%; P = ,002). We did not observe any differences in postoperative complications. The mean hospital stay was significantly lower in Va-PCNL with 1,7 ± 1,9 days per patient compared with 2,7 ± 1,5 days in the Mini-PCNL group (P = ,001). There were no differences in SFR at 3 months between Va-PCNL (71,9%) and Mini-PCNL (71,8%; P =v ,848)., Conclusion: Patients treated with Va-PCNL had comparable results to Mini-PCNL, showing equal SFR with similar infectious complications rates. Potential benefits of Va-PCNL include shorter operative time and postoperative stay., (Copyright © 2023 AEU. Published by Elsevier España, S.L.U. All rights reserved.)

Brain-computer interfaces (BCIs) establish a direct communication channel between the human brain and external devices. Among various methods, electroencephalography (EEG) stands out as the most popular choice for BCI design due to its non-invasiveness, ease of use, and cost-effectiveness. This paper aims to present and compare the accuracy and robustness of an EEG system employing one or two channels. We present both hardware and algorithms for the detection of open and closed eyes. Firstly, we utilize a low-cost hardware device to capture EEG activity from one or two channels. Next, we apply the discrete Fourier transform to analyze the signals in the frequency domain, extracting features from each channel. For classification, we test various well-known techniques, including Linear Discriminant Analysis (LDA), Support Vector Machine (SVM), Decision Tree (DT), or Logistic Regression (LR). To evaluate the system, we conduct experiments, acquiring signals associated with open and closed eyes, and compare the performance between one and two channels. The results demonstrate that employing a system with two channels and using SVM, DT, or LR classifiers enhances robustness compared to a single-channel setup and allows us to achieve an accuracy percentage greater than 95% for both eye states.

Introduction: Erectile dysfunction (ED) is one of the most prevalent urological diseases, but there is limited data about the quality of its information in social networks. The aim of our study was to assess the quality of ED information contained in YouTube videos., Material and Methods: Descriptive study of the first 50 Spanish-language videos, published on YouTube, evaluated by three urologists. We used two validated questionnaires: PEMAT (Patient Education Materials Assessment Tool) and DISCERN. Videos were classified according to DISCERN score into poor or moderate-good quality., Results: The median time duration was 2.42minutes (0.15-3.58), 94,197 views (2,313-3,027,890), 682.5 «likes» (0-54,020) and 39 «dislikes» (0-2843). The median of PEMAT score was 29% (9%-95.5%) in understandability and 29% (0-95.5%) in actionability. According to DISCERN score 27 videos (57.4%) had poor quality and 20 (42.6%) moderate-good quality. There were no significant differences between the two groups in time duration, views, «likes» or «dislikes». There were differences in PEMAT score in understandability and actionability. The 86.7% of the moderate-good quality videos were starred by health care provider (P=.001). Also, the 85.7% of videos that describes treatment had moderate-good quality (P=.001). The 84% of the non-medical videos had a poor quality (P=.001)., Conclusion: Most ED videos on YouTube have poor quality. The highest quality videos are those made by professionals, although they are not the most viewed. It would be important to develop measures to prevent the spread of misinformation among social network users., (Copyright © 2023 Asociación Española de Andrología, Medicina Sexual y Reproductiva. Publicado por Elsevier España, S.L.U. All rights reserved.)

The textile industry produces harmful effluents that are discharged into the environment, damaging the aquatic and other ecosystems. A yeast-based solution for decolorization of textile industrial wastewater was produced and evaluated. Three yeast strains, Candida parapsilosis (HOMOGS20B), Yarrowia lipolytica (HOMOGST27AB) and Candida pseudoglaebosa (LIIIS36B), isolated from a textile wastewater treatment plant and previously selected for their dye decolorization capacity, were freeze-dried. Additionally, Yarrowia lipolytica (HOMOGST27AB) was also spray-dried. Skim milk powder and maltodextrin were used as cell protectors, and the freeze-dried products were stored at cold (4 °C) and room temperature for 210 days. The viability of the yeast cells and their decolorization capacity over time were assessed. Dried yeast cells maintained their viability, and decolorization capacity for at least 90 days of storage after spray- and freeze-drying with both cell-protecting agents. The dried yeast-based solution for decolorizing textile industrial wastewater combines stability, efficiency, and convenience of production for application in real industrial facilities., 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 British Mycological Society. Published by Elsevier Ltd. All rights reserved.)

Poor aqueous solubility, stability and bioavailability of interesting bioactive compounds is a challenge in the development of bioactive formulations. Cellulose nanostructures are promising and sustainable carriers with unique features that may be used in enabling delivery strategies. In this work, cellulose nanocrystals (CNC) and cellulose nanofibers were investigated as carriers for the delivery of curcumin, a model liposoluble compound. Nanocellulose modification with the surfactant cetyltrimethylammonium bromide (CTAB), tannic acid and decylamine (TADA), and by TEMPO-mediated oxidation were also tested and compared. The carrier materials were characterized in terms of structural properties and surface charge, while the delivery systems were evaluated for their encapsulation and release properties. The release profile was assessed in conditions that mimic the gastric and intestinal fluids, and cytotoxicity studies were performed in intestinal cells to confirm safe application. Modification with CTAB and TADA resulted in high curcumin encapsulation efficiencies of 90 and 99%, respectively. While no curcumin was released from TADA-modified nanocellulose in simulated gastrointestinal conditions, CNC-CTAB allowed for a curcumin-sustained release of ca. 50% over 8 h. Furthermore, the CNC-CTAB delivery system showed no cytotoxic effects on Caco-2 intestinal cells up to 0.125 g/L, meaning that up to this concentration the system is safe to use. Overall, the use of the delivery systems allowed for the reduction in the cytotoxicity associated with higher curcumin concentrations, highlighting the potential of nanocellulose encapsulation systems.

Dyed effluents from textile industry are toxic and difficult to treat by conventional methods and biotechnological approaches are generally considered more environmentally friendly. In this work, yeast strains Candida parapsilosis, Yarrowia lipolytica and Candida pseudoglaebosa, isolated from wastewater treatment plants, were tested for their ability to decolorize textile dyes. Both commercial textile synthetic dyes (reactive, disperse, direct, acid and basic) and simulated textile effluents (a total of 32 solutions) were added to a Normal Decolorization Medium along with the yeast (single strains and consortia) and the decolorization was evaluated spectrophotometrically for 48-72 h. Yeasts were able to perform decolorization through adsorption and biodegradation for 28 of the dyes and simulated effluents by more than 50%. Y. lipolytica and C. pseudoglaebosa presented the best results with a true decolorization of reactive dyes, above 90% at 100 mg l -1 , and simulated effluents at 5 g l -1 of concentration. Enzyme production was evaluated: oxidoreductase was found in the three yeasts, whereas tyrosinase was only found in Y. lipolytica and C. pseudoglaebosa. Y. lipolytica and C. pseudoglaebosa are a potential biotechnological tool for dye degradation in textile wastewaters, especially those containing reactive dyes and a promising tool to integrate in bioremediation solutions, contributing to circular economy and eco sustainability in the water sector since the treated water could possibly be reused for irrigation., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Human-Machine Interfaces (HMI) allow users to interact with different devices such as computers or home elements. A key part in HMI is the design of simple non-invasive interfaces to capture the signals associated with the user's intentions. In this work, we have designed two different approaches based on Electroencephalography (EEG) and Electrooculography (EOG). For both cases, signal acquisition is performed using only one electrode, which makes placement more comfortable compared to multi-channel systems. We have also developed a Graphical User Interface (GUI) that presents objects to the user using two paradigms-one-by-one objects or rows-columns of objects. Both interfaces and paradigms have been compared for several users considering interactions with home elements.

Grape pomace (GP) is a major by-product from the wine industry, known for its bioactive compounds and their impact upon gastrointestinal (GI) health. However, bioaccessibility is often poor due to their degradation during digestion. This work aimed to encapsulate bioactive GP extract (GPE) into chitosan (CS) and alginate (Alg) nanoparticles (NPs) to mitigate degradation in the GI tract. Alg and CS NPs were optimized using a rotatable central composite design and NPs were characterized for their size, polydispersity, zeta potential and total phenolics (TP) association efficiency. The best formulations showed sizes ranging 523-853 nm, polydispersity indexes of 0.11-0.36, zeta potential of -15.0-14.9 mV and TP association efficiencies of 68 and 65%. FTIR confirmed that there was no formation of new chemical groups after association of the polymers with GPE. Both formulations improved the bioaccessibility of different phenolics following in vitro GI digestion, leading to increased antioxidant and antimicrobial activities. Moreover, the permeability of bioactive compounds through a Caco-2/HT29-MTX co-culture was reduced, suggesting a higher residence time in the intestine. Cy5.5 was used for tracking the CS NPs, which did not affect the metabolic activity of Caco-2 and HT29-MTX cells. Confocal microscopy images confirmed the adsorption of NPs to the cellular layer and suggested a reduction of the tight junction protein occludin when cells were incubated with Cy5.5-CS in solution. This study suggests that encapsulation of GPE can offer protection against along the GI tract and improve its biological activity with significant impact for oral delivery applications, including functional foods., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Background and Purpose: Although transcranial Doppler detects microembolic signals (MES) in numerous settings, the practical significance of such findings remains unclear., Methods: Clinical information from ischemic stroke or transient ischemic attack patients (n = 248) who underwent embolic monitoring from January 2015 to December 2018 was obtained., Results: MES were found in 15% of studies and ischemic recurrence was seen in 11% of patients (over 7 ± 6 days). Patients with MES had more lacunes than those without MES (1 ± 3 vs. 1 ± 2, P = .016), were more likely to have ischemic recurrence (37% vs. 6%, P < .001), undergo a future revascularization procedure (26% vs. 10%, P = .005), have a longer length of stay (9 vs. 4 days, P = .043), and have worse functional disability at discharge (modified Rankin Scale 3-6, 66% vs. 34%, P < .001). After controlling for several relevant cofactors, patients with MES were more likely to have ischemic recurrence (HR 4.90, 95% CI 2.16-11.09, P < .001), worse functional disability (OR 3.31, 95% CI 1.22-8.99, P = .019), and longer length of stays (β = .202, P < .001)., Conclusions: MES may help to risk stratify patients as their presence is associated with ischemic recurrence and worse outcomes., (© 2020 American Society of Neuroimaging.)

In this work, we develop open source hardware and software for eye state classification and integrate it with a protocol for the Internet of Things (IoT). We design and build the hardware using a reduced number of components and with a very low-cost. Moreover, we propose a method for the detection of open eyes (oE) and closed eyes (cE) states based on computing a power ratio between different frequency bands of the acquired signal. We compare several real- and complex-valued transformations combined with two decision strategies: a threshold-based method and a linear discriminant analysis. Simulation results show both classifier accuracies and their corresponding system delays.

The buccal route is considered patient friendly due to its non-invasive nature and ease of administration. Such delivery route has been used as an alternative for the delivery of drugs that undergo first-pass metabolism or are susceptible to pH and enzymatic degradation, such as occurs in the gastrointestinal tract. However, the drug concentration absorbed in the buccal mucosa is often low to obtain an acceptable therapeutic effect, mainly due to the saliva turnover, tongue and masticatory movements, phonation, enzymatic degradation and lack of epithelium permeation. Therefore, the encapsulation of drugs into nanoparticles is an important strategy to avoid such problems and improve their buccal delivery. Different materials from lipids to natural or synthetic polymers and others have been used to protect and deliver drugs in a sustained, controlled or targeted manner, and enhance their uptake through the buccal mucosa improving their bioavailability and therapeutic outcome. Overall, the main aim of this review is to perform an overview about the nanotechnological approaches developed so far to improve the buccal delivery of drugs. Herein, several types of nanoparticles and delivery strategies are addressed, and a special focus on pipeline products is also given., (Copyright © 2020 Elsevier B.V. All rights reserved.)

A method for channel estimation in wideband massive MIMO systems using hybrid digital analog architectures is developed. The proposed method is useful for FDD at either sub-6 GHz or mmWave frequency bands and takes into account the beam squint effect caused by the large bandwidth of the signals. To circumvent the estimation of large channel vectors, the posed algorithm relies on the slow time variation of the channel spatial covariance matrix, thus allowing for the utilization of very short training sequences. This is possibledue to the exploitation of the channel structure. After identifying the channel covariance matrix, the channel is estimated on the basis of the recovered information. To that end, we propose a novel method that relies on estimating the tap delays and the gains as sociated with each path. As a consequence, the proposed channel estimator achieves low computational complexity and significantly reduces the training overhead. Moreover, our numerical simulations show better performance results compared to the minimum mean-squared error solution.

Whey-derived alpha-casozepine bioactive peptide (YLGYLEQLLR) was associated with previously optimized guar-gum film-PLGA nanoparticles, aiming to increase both stability across gastrointestinal tract and permeability across absorptive epithelia. Oral films associated with nanoparticles (FNp) enhance buccal absorption along with protection of carried bioactive molecules that are swallowed, with inherent increase of bioavailability. None of developed formulations induced significant loss of cell viability. Permeability across both buccal and intestinal cell barriers was enhanced when alpha-casozepine was carried by FNp system, when compared with film and nanoparticles alone, in a simulated gastrointestinal tract environment. Moreover, differences in permeability profile across buccal and intestinal epithelia were in accordance with the slower erosion of PLGA nanoparticles in a media of neutral pH, resembling oral cavity conditions, and a faster erosion in acidic conditions, as occurs in stomach, as observed by a continuous analysis of nanoparticle morphology over 980 min by atomic force microscopy. Additionally, apparent permeability of alpha-casozepine across TR146 human buccal carcinoma cells and Caco-2/HT29-MTX co-culture, carried by FNp was indeed superior when compared with peptide loaded in PLGA nanoparticles and in films alone or with free peptide control solution. Both FNp and PLGA nanoparticles alone enhanced the permeability of relaxing peptide compared with guar-gum films alone. An increased tongue adhesion when PLGA nanoparticles were added to the guar-gum films was also observed. Developed formulations improved both buccal an intestinal absorption of carried bioactive molecules without compromising cell viability., (Copyright © 2019. Published by Elsevier B.V.)

Background: Arterial baroreflex (BR) and cerebral autoregulation (CA) are two major regulatory mechanisms that maintain constant cerebral perfusion. Little is known about the interplay between these mechanisms, particularly when considering the effects of ageing or sex., Purpose: We studied the relationship between dynamic CA and BR sensitivity (BRS) in healthy subjects by sex and in different age strata., Methods: 95 healthy adults (52% female), 20-80 years-old, were recruited. Arterial blood pressure (Finometer), 3-lead electrocardiogram and cerebral blood flow velocity in middle cerebral arteries (transcranial Doppler) were monitored. We assessed CA by transfer function analysis and BRS in frequency and time domain., Results: With increasing age, BRS diminished (ANCOVA R 2  = 0.281, p < 0.001) but CA parameters did not change significantly (p > 0.05). Overall, there was an inverse relationship between the efficacy of BRS and CA low-frequency gain [multivariate linear regression β = 0.41 (0.31; 0.61), p < 0.001]. However, this association suffers changes with ageing: in older subjects BRS and CA were not correlated [β = 0.10 (- 0.41; 0.62), p = 0.369]. Instead, decreasing systolic blood pressure correlated with less efficient CA [lower CA low-frequency gain β = - 0.02 (- 0.03; - 0.02), p = 0.003]. Sex did not affect BRS and CA relationship., Conclusions: Cerebral blood supply is governed by a tuned balance between BR and CA which is lost with age as BRS decreases dramatically. Low systolic blood pressure values might be harmful to older subjects as they might reduce the ability to keep cerebral blood flow tightly controlled.

Trees in urban areas face harsh environmental conditions. Ectomycorrhizal fungi (EcM) form a symbiosis with many tree species and provide a range of benefits to their host through their extraradical hyphal network. Although our understanding of the environmental drivers and large scale geographical variation of EcM communities in natural ecosystems is growing, our knowledge of EcM communities within and across urban areas is still limited. Here, we characterized EcM communities using Illumina miseq sequencing on 175 root samples of the urban tree Tilia tomentosa from three European cities, namely Leuven (Belgium), Strasbourg (France) and Porto (Portugal). We found strong differences in EcM richness and community composition between cities. Soil acidity, organic matter and moisture content were significantly associated with EcM community composition. In agreement, the explained variability in EcM communities was mostly attributed to general soil characteristics, whereas very little variation was explained by city and heavy metal pollution. Overall, our results suggest that EcM communities in urban areas are significantly associated with soil characteristics, while heavy metal pollution and biogeography had little or no impact. These findings deliver new insights into EcM distribution patterns in urban areas and contribute to specific inoculation strategies to improve urban tree vitality.

Oral administration of proteins and peptides still is a challenging task to overcome due to low permeability through absorptive epithelia, degradation and metabolism that lead to poor bioavailability. Attempting to overcome such limitations, an antihypertensive peptide derived from whey protein, with KGYGGVSLPEW sequence, was incorporated for the first time into polymeric nanoparticles. An experimental design was followed in order to optimize drug-loading, association efficiency, mean particle size, zeta-potential and polydispersity index of a formulation of poly(lactic-co-glycolic acid) (PLGA) nanoparticles as carriers for bioactive peptides. In sequence, peptide-loaded PLGA nanoparticles were incorporated in a guar-gum film matrix, resulting in a combined delivery system aiming to promote slow release and permeation across buccal epithelium. Neither PLGA nanoparticles, guar-gum films nor the conjugation of PLGA nanoparticles and guar-gum films (GfNp) significantly compromised in vitro TR146 human buccal carcinoma cell line viability after 12 h contact, as assessed by 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide reduction assay (MTT). In vitro release assay for developed formulations allowed to conclude that the combination of orodispersible film and nanoparticles granted a slower release of AhP when compared with PLGA or guar-gum films alone or with control. GfNp offered more effective, synergistic, in vitro permeation of TR146 cell multilayer in comparison with guar-gum films or PLGA nanoparticles alone. The combination of PLGA nanoparticles with guar-gum films represent a suitable alternative to conventional per os delivery systems, leading to an increased buccal permeability of carried antihypertensive peptide., (Copyright © 2018 Elsevier B.V. All rights reserved.)

The association of alginate beads and guar-gum films in a single delivery system was idealized to promote a more effective buccal and oral delivery of bioactive molecules. A response surface method (experimental design approach) was performed to obtain optimal formulations of alginate beads to be incorporated into guar gum oral films as combined buccal and oral delivery systems for caffeine delivery. The combined formulation was further characterized regarding physicochemical properties, drug release, cell viability and buccal permeability. Beads average size, determined by dynamic light scattering (DLS), was of 3.37 ± 6.36 μm. Film thickness was set to 62 μm. Scanning electron microscopy micrographs revealed that beads were evenly distributed onto the film matrix and beads size was in accordance to data obtained from DLS analysis. Evaluation of Fourier-transform infrared spectra did not indicate the formation of new covalent bonds between the matrix of guar-gum films, alginate beads and caffeine. In vitro release assays by dialysis membrane allowed understanding that the combination of guar-gum films and alginate beads assure a slower release of caffeine when compared with the delivery profile of free caffeine from alginate beads or guar-gum films alone. MTT assay, performed on human buccal carcinoma TR146 cell line, allowed concluding that neither guar-gum film, alginate beads nor guar-gum film incorporated into alginate beads significantly compromised cell viability after 12 h of exposure. As demonstrated by in vitro permeability assay using TR146 human buccal carcinoma cell lines, combination of guar-gum films and alginate beads also promoted a slower release and, thus, lower apparent permeability (1.15E-05 ± 3.50E-06) than for caffeine solution (2.68E-05 ± 7.30E-06), guar-gum film (3.12E-05 ± 4.70E-06) or alginate beads (2.01E-05 ± 3.90E-06). The conjugation of alginate beads within an orodispersible film matrix represents an effective oral/buccal delivery system that induces a controlled release along with an enhanced intimate contact with cell layers that may promote higher in vivo bioavailability of carried drugs., (Copyright © 2018. Published by Elsevier Ltd.)

Bioactive proteins and peptides have been used with either prophylactic or therapeutic purposes, presenting inherent advantages as high specificity and biocompatibility. Nanocarriers play an important role in the stabilization of proteins and peptides, offering enhanced buccal permeation and protection while crossing the gastrointestinal tract. Moreover, preparation of nanoparticles as oral delivery systems for proteins/peptides may include tailored formulation along with functionalization aiming bioavailability enhancement of carried proteins or peptides. Oral delivery systems, namely buccal delivery systems, represent an interesting alternative route to parenteric delivery systems to carry proteins and peptides, resulting in higher comfort of administration and, therefore, compliance to treatment. This paper outlines an extensive overview of the existing publications on proteins/peptides oral nanocarriers delivery systems, with special focus on buccal route. Manufacturing aspects of most commonly used nanoparticles for oral delivery (e.g. polymeric nanoparticles using synthetic or natural polymers and lipid nanoparticles) advantages and limitations and potential applications of nanoparticles as proteins/peptides delivery systems will also be thoroughly addressed., (Copyright © 2018 Elsevier Inc. All rights reserved.)

The computation of a set constituted by few vertices to define a virtual backbone supporting information interchange is a problem that arises in many areas when analysing networks of different natures, like wireless, brain, or social networks. Recent papers propose obtaining such a set of vertices by computing the connected dominating set (CDS) of a graph. In recent works, the CDS has been obtained by considering that all vertices exhibit similar characteristics. However, that assumption is not valid for complex networks in which their vertices can play different roles. Therefore, we propose finding the CDS by taking into account several metrics which measure the importance of each network vertex e.g., error probability, entropy, or entropy variation (EV)., Competing Interests: The authors declare no conflict of interest.

Chitosan is a biocompatible, bioactive, non-toxic polymer that due to these characteristics has been widely used as a carrier for targeted delivery of bioactive molecules. In recent years, and considering that chitosan has a strong antimicrobial potential, the scientific community's focus has shifted onto the possible antimicrobial activity of chitosan nanoparticles. With this in mind, the aim of this work was to produce low molecular weight chitosan nanoparticles, through the ionic gelation method and characterize their potential biological activity against three staphylococci (MSSA, MRSA and MRSE) in planktonic and sessile environments. The chitosan nanoparticles produced had an average size of 244±12nm, an average charge of 17.3±1.4mV and had a MIC of 1.25mg/mL for all tested microorganisms. Bactericidal activity was only registered for MSSA and MRSA with the time-inhibition curves showing bactericidal activity within 1h. Assays regarding chitosan nanoparticles' impact upon sessile populations showed that they were effective in preventing MRSE adhesion and highly effective in reducing MRSA and MSSA biofilm formation., (Copyright © 2017. Published by Elsevier B.V.)

Background: Atrial fibrillation (AF) is a major risk factor for ischemic stroke (IS). Patients with AF may undergo preventive therapy. Although the AF impact in the clinical burden of IS has been studied, information is lacking in Southern Europe and there are no studies about the impact in potential years of life lost. Moreover, no nationwide or long-term study analyzed the economic burden of IS stratified by AF., Objective: To study the impact of AF in the clinical and economic burden of IS., Methods: We conducted a retrospective study using nationwide administrative data for all public hospitalizations in mainland Portugal from 2000 to 2014. We considered IS hospitalizations stratified by the presence of AF as secondary diagnosis., Results: Of the total 275,173 IS hospitalizations, 22.6% reported AF. The total number of IS hospitalizations increased from 14,836 in 2000 to 19,561 in 2014 (32% increase), with an increase of 138% in the AF group (from 2,411 to 5,727). In-hospital mortality decreased from 13.6% to 11.5% and was consistently higher in the AF group (17.3% vs. 11.1%). Mean charges were also higher in the AF group (€2297 vs. €2191). Age-adjusted potential years of life lost rate was higher in the group without AF (39.6 vs. 7.5)., Conclusions: AF-associated IS hospitalizations more than doubled in the studied 15-year period. Also, AF was responsible for higher in-hospital mortality and hospitalization charges. These facts highlight the need for early detection of AF and preventive treatment to limit IS occurrence, its associated burden, and poorer health outcomes., (Copyright © 2017 International Society for Pharmacoeconomics and Outcomes Research (ISPOR). Published by Elsevier Inc. All rights reserved.)

An experimental design was established in order to optimize the mechanical properties of two oral film formulations intended for oral delivery of bioactive compounds. Carboxymethylcellulose (CMC) and gelatin type A (GelTA) were selected as polymeric matrix. Scanning electron microscopy revealed that caffeine crystals were homogeneously dispersed onto oral film matrix. Fourier-transform infrared analysis did not indicate formation of new chemical entities. USP modified dissolution assay revealed that GelTA was more effective in controlling caffeine release since maximum caffeine release (97.4%±0.95) after 20min. On the other hand, CMC is better indicated for immediate release since maximum caffeine release (81.1%±2.14) occurred after 4min. Simulation of gastrointestinal tract with ex vivo permeability assay was in accordance with USP dissolution assay (42.0%±7.79 and 15.3%±4.0 of caffeine released from CMC and GelTA oral films (OF), respectively, permeated porcine intestinal mucosa after 120min). CMCOF and GelTAOF optimized formulations represent two suitable oral delivery systems for immediate and controlled release, respectively., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Calcium hydroxyapatite Ca 10 (PO 4 ) 6 (OH) 2 (HAp) is a material widely used in biomedicine, for bone implants manufacture, due to its biocompatibility. HAp has also application for environmental remediation, as it can be employed as metal removal; moreover, it has the capability of effectively adsorbing organic molecules its surface. In recent years, the photocatalytic properties of HAp have been investigated; indeed several studies report of HAp used as photocatalyst, either on its own or combined with other photocatalytic materials. Although in the majority of cases the activity was induced by UV light, some reports of visible light-activated materials were reported. Here we present a critical review of the latest developments for HAp-based photocatalysts; the materials discussed are undoped single phase HAp, doped HAp and HAp-containing composites. For undoped single phase HAp, the possible surface treatment and lattice defects which can lead to a photoactive material are discussed. Considering doped HAp, the use of Ti 4+ (the most common dopant) is described, with particular attention to the effects that this metal have on the characteristics of the material (i.e. crystallinity) and on its photocatalytic behaviour. The use of other dopants is also discussed. For the multiphasic materials, the combination of HAp with other photocatalysts is discussed, mainly but not only with titanium dioxide TiO 2 . Overall, HAp is a compound with high potential as photocatalyst; this property, combined with its capability for heavy metal removal, makes it a multifunctional material for environmental remediation. As future perspectives, further studies, based on the results obtained until present, should be performed, to improve the performance of the materials and/or shift the band gap into the visible. The use of other dopants and/or the combination with other photocatalysts, for instance, are features which is worth exploring., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Objectives: Surgical restoration of pelvic floor anatomy in pelvic organ prolapse (POP) should avoid extensive areas that may injure healthy tissues and lead to scar fibrosis producing dysfunctional rigidity. Laparoscopic sacrocolpopexy corrects POP by lifting pelvic elements with a minimally invasive procedure.Various current strategies and approaches make it a diverse procedure., Methods: We performed a bibliographic review on the published experience about abdominal sacrocolpopexy over the last 20 years. We analyze the philosophy of each technique, advantages, disadvantages, and results., Results: Among current theories that explain how is the anatomy and function of the pelvic floor, the comprehensive theory of the pelvic floor announces that anatomical deviation produces pelvic dysfunction (diagnostic algorithm)(1). Surgical invasion with reconstructive purposes may hide results if the true physiopathology of the defects found is not known. Although current diagnostic tools cannot solve the problem as a whole, results of different techniques are much better than those of previous times., Conclusions: Laparoscopic sacrocolpopexy is an adequate strategy that restores anatomy in POP, mainly apical, and function with minimal invasion. Technical development and better knowledge of the female pelvis functional anatomy is enabling today a more precise understanding of its physiopathology, so it facilitates the design of site-specific operations. Various surgical approaches and techniques are offered depending on the defect, where combined procedures give good solutions. It is necessary to have an appropriate surgical training to obtain the better competence, and a safe and correct repair.

Layered (Bi 1-x In x ) 2 Te 3 -In 2 Te 3 (x = 0.075) composites of pronounced anisotropy in structure and thermoelectric properties were produced by zone melting and subsequent coherent precipitation of In 2 Te 3 from a (Bi 1-x In x ) 2 Te 3 (x > 0.075) matrix. Employing solid state phase transformation, the Bi 2 Te 3 /In 2 Te 3 interface density was tuned by modifying the driving force for In 2 Te 3 precipitation. The structure-property relationship in this strongly anisotropic material is characterized thoroughly and systematically for the first time. Unexpectedly, with increasing Bi 2 Te 3 /In 2 Te 3 interface density, an increase in electrical conductivity and a decrease in the absolute Seebeck coefficient were found. This is likely to be due to electron accumulation layers at the Bi 2 Te 3 /In 2 Te 3 interfaces and the interplay of bipolar transport in Bi 2 Te 3 . Significantly improved thermoelectric properties of Bi 2 Te 3 -In 2 Te 3 composites as compared to the single phase (Bi 1-x In x ) 2 Te 3 solid solution are obtained.

The development of innovative, safe and non-photocatalytic sunscreens is urgently needed, as it is essential to have sunscreen filters offering appropriate UV protection without damaging the environment and/or generating free radicals when in contact with the skin. Hydroxyapatite (Ca 10 (PO 4 ) 6 (OH) 2 , HAp) when substituted with iron has UV protection properties and is not photocatalytic; HAp was used to make a sunscreen filter by treating cod fish bones in an iron-containing solution, and then calcining them at 700°C. Here we present a systematic and advanced study on this material, to obtain a sunscreen with improved UV absorbing properties. Bones were treated with three different iron salts - Fe(II) chloride, Fe(II) lactate and Fe(III) nitrate - under various pH conditions. Results showed that Fe(II) chloride in basic pH led to the most effective iron inclusion. High energy ball milling or ultrasound were investigated to increase surface area and corresponding UV absorption; high energy ball milling treatment led to the best optical properties. The optimum powders were used to formulate UV protection creams, which showed Sun Protection Factor (SPF) values significantly superior to the control cream (up to 4.1). Moreover the critical wavelength (λ crit ) was >370nm (388-389nm) and UVA/UVB ratios were very close to 1. With these properties these sunscreens can be classified as broad UV protectors. Results also showed that combining these powders with other sunscreens (i.e. titanium dioxide), a synergic effect between the different components was also observed. This investigation showed that HAp-based sunscreens of marine origin are a valid alternative to commercial products, safe for the health of the customers and, being non-photocatalytic, do not pose a threat to the environment., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Soil heavy metal contamination resulting from mining activities constitutes a major environmental problem worldwide. The spread of heavy metals is often facilitated by scarce vegetation cover, so there is an urgent need to improve plant survival and establishment in these metalliferous areas. This study is aimed at the isolation and analysis of the phylogenetic relationship of culturable bacteria from the rhizosphere of metallophyte plants growing in the Kettara mine, in Marrakech, in order to select plant growth-promoting rhizobacteria (PGPR), which could be used in assisted-phytoremediation. Bacterial isolates were grouped by random amplified polymorphic DNA analysis and identified by 16S rRNA gene sequencing. Strains were further characterized for the production of plant growth-promoting (PGP) substances, such as NH 3 , siderophores, indol-3-acetic acid (IAA), hydrogen cyanide, and extracellular enzymes, for ACC-deaminase activity, their capacity to solubilize phosphate, and for their tolerance to heavy metals and acidic pH. Rhizosphere soils were highly contaminated with Cu and Zn and presented low fertility. Phylogenetic analysis showed that the rhizobacteria were affiliated to three major groups: γ-Proteobacteria (48 %), β-Proteobacteria (17 %), and Bacilli (17 %). The most represented genera were Pseudomonas (38 %), Bacillus (10 %), Streptomyces (10 %), and Tetrathiobacter (10 %). Overall, rhizobacterial strains showed an ability to produce multiple, important PGP traits, which may be helpful when applied as plant growth promoter agents in contaminated soils. PGPR were also able to withstand high levels of metals (up to 2615.2 mg Zn l -1 , 953.29 mg Cu l -1 , and 1124.6 mg Cd l -1 ) and the order of metal toxicity was Cd > Cu > Zn. The rhizobacterial strains isolated in the present study have the potential to be used as efficient bioinoculants in phytoremediation strategies for the recovery of Kettara mine soils.

5,10,15,20-tetrakis(1-methylpyridinium-4-yl)-porphyrin tetra-iodide (TMPyP), a potent water-soluble photosensitizer (PS) used in antimicrobial applications, was encapsulated into poly(lactic-co-glycolic acid) (PLGA) nanoparticles (TMPyP-PLGA) for topical delivery purposes. Nanoparticles resulted in a mean particle size around 130nm, narrow polydispersity index (PdI), spherical morphology and association efficiency up to 93%. Free TMPyP and TMPyP-PLGA nanoparticles were incorporated into Carbopol(®) hydrogels, resulting in controlled TMPyP release of about 60% and 20% after 4.5h, respectively. Critical properties such as appearance, clarity, viscosity and pH were maintained over time, as hydrogels were stable during 6 months at 4°C, 25°C/60% RH and 40°C/75% RH. For photodynamic applications, the photoproduction of singlet oxygen from these hydrogels was quite efficient being both formulations very photostable after 20min. No TMPyP permeation through pig ear skin was observed after 24h, and histological assays did not show relevant damages in surrounding tissues. All these excellent characteristics make them promising platforms for photodynamic applications through topical clinical use., (Copyright © 2016 Elsevier B.V. All rights reserved.)

This work provides a system capable of obtaining simultaneous inductive signatures of vehicles traveling on a roadway with minimal cost. Based on Time-Division Multiplexing (TDM) with multiple oscillators, one for each inductive loop, the proposed system detects the presence of vehicles by means of a shift in the oscillation period of the selected loop and registers the signature of the detected vehicles by measuring the duration of a fixed number of oscillator pulses. In order to test the system in an actual environment, we implement a prototype that we denote as SiDIVS (Simple Detection of Inductive Vehicle Signatures) and acquire different vehicle inductive signatures under real scenarios. We also test the robustness of the detector by simulating the effect of noise on the signature acquisition.

The use of heavy metals (HM) contaminated soils to grow energy crops can diminish the negative impact of HM in the environment improving land restoration. The effect of two PGPR (B1--Chryseobacterium humi ECP37(T) and B2--Pseudomonas reactans EDP28) and an AMF (F--Rhizophagus irregularis) on growth, Cd and Zn accumulation, and nutritional status of energy maize plants grown in a soil collected from an area adjacent to a Portuguese mine was assessed in a greenhouse experiment. Both bacterial strains, especially when co-inoculated with the AMF, acted as plant growth-promoting inoculants, increasing root and shoot biomass as well as shoot elongation. Cadmium was not detected in the maize tissues and a decrease in Zn accumulation was observed for all microbial treatments in aboveground and belowground tissues--with inoculation of maize with AMF and strain B2 leading to maximum reductions in Zn shoot and root accumulation of up to 48 and 43%, respectively. Although microbial single inoculation generally did not increase N and P levels in maize plants, co-inoculation of the PGPR and the AMF improved substantially P accumulation in roots. The DGGE analysis of the bacterial rhizosphere community showed that the samples inoculated with the AMF clustered apart of those without the AMF and the Shannon-Wiener Index (H') increased over the course of the experiment when both inoculants were present. This work shows the benefits of combined inoculation of AMF and PGPR for the growth energy maize in metal contaminated soils and their potential for the application in phytomanagement strategies.

Fluoroquinolones constitute a group of emerging pollutants and their occurrence in different environmental compartments is becoming object of increasing public concern due to their ecotoxicological effects and the potential to develop resistant bacteria. This study aimed to investigate the biodegradation of moxifloxacin (MOX), for which studies in the literature are very scarce. An activated sludge (AS) consortium and three bacterial strains able to degrade fluoroaromatic compounds - strains F11, FP1 and S2 - were tested. Biodegradation studies were conducted using acetate as a bulk carbon source. Strain F11 showed the highest biodegradation capacity, being able to completely consume and dehalogenate 7.5 μM of the target antibiotic when daily co-supplemented with acetate present as a readily degradable organic substrate in wastewaters. MOX could be used by strain F11 as a sole nitrogen source but the presence of an external nitrogen source in the culture medium was essential for complete biodegradation. Strain F11 was capable of completely consuming MOX in a range between 2 and 11 μM, although stoichiometric fluoride release was not obtained for the highest tested concentration. The antibacterial activity of residual MOX and of the metabolic products potentially resultant from the biodegradation process was investigated by agar diffusion tests, demonstrating that MOX biodegradation is associated with the elimination of the antibacterial properties of the target antibiotic and of the produced metabolites, which is an important result, as the activity of antibiotics and/or their metabolites in the environment, even at low levels, may lead to the development of resistant bacterial strains. Overall, the results obtained in this study suggest that strain F11 is a promising microorganism for the treatment of waters contaminated with MOX, where it could be used for bioaugmentation/bioremediation purposes. To the best of our knowledge, this is the first study reporting complete removal and dehalogenation of MOX by a single microorganism., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Green roofs (GRs) are becoming a trend in urban areas, favouring thermal performance of buildings, promoting removal of atmospheric pollutants, and acting as possible water collection spots. Rainwater harvesting systems in buildings can also contribute to the management of stormwater runoff reducing flood peaks. These technologies should be enhanced in Mediterranean countries where water scarcity is increasing and the occurrence of extreme events is becoming very significant, as a result of climate change. An extensive pilot GR with three aromatic plant species, Satureja montana, Thymus caespititius and Thymus pseudolanuginosus, designed to study several parameters affecting rainwater runoff, has been in operation for 12 months. Physico-chemical analyses of roof water runoff (turbidity, pH, conductivity, NH4(+), NO3(-), PO4(3-), chemical oxygen demand) have shown that water was of sufficient quality for non-potable uses in buildings, such as toilet flushing. An innovative approach allowed for the development of an expression to predict a 'monthly runoff coefficient' of the GR system. This parameter is essential when planning and designing GRs combined with rainwater harvesting systems in a Mediterranean climate. This study is a contribution to improving the basis for the design of rainwater harvesting systems in buildings with extensive GRs under a Mediterranean climate.

In this work we study opinion formation in a population participating in a public debate with two distinct choices. We consider three distinct mechanisms of social interactions and individuals' behavior: conformity, nonconformity, and inflexibility. The conformity is ruled by the majority-rule dynamics, whereas the nonconformity is introduced in the population as an independent behavior, implying the failure of attempted group influence. Finally, the inflexible agents are introduced in the population with a given density. These individuals present a singular behavior, in a way that their stubbornness makes them reluctant to change their opinions. We consider these effects separately and all together, with the aim to analyze the critical behavior of the system. We perform numerical simulations in some lattice structures and for distinct population sizes. Our results suggest that the different formulations of the model undergo order-disorder phase transitions in the same universality class as the Ising model. Some of our results are complemented by analytical calculations.

Introduction: Urinary lithiasis is the main urologic cause of emergency treatment in adult patient. In the past years, the incidence in children population has increased. However, literature about the use of alpha-1 adrenergic blockers in pediatric population with distal ureterolithiasis is still scarce. The drug acts by decreasing ureter contractions, especially in the distal portion, facilitating calculus expulsion., Objective: This review has the objective to evaluate the use of alpha-1 adrenergic blockers as medical expulsive treatment in children with distal ureterolithiasis., Evidence Acquisition: An electronic literature search was performed using the MEDLINE, COCHRANE, and LILACS databases. We further searched manually the references of the primary studies. Searches were concluded on October 4th, 2014. Articles were selected, independently and in pairs, by the respective titles and summaries. Any divergence was resolved by consensus., Evidence Synthesis: Alpha-1 adrenergic antagonists increased the probability of calculus expulsion by 27% (NNT=4). Calculi smaller than 5mm, increased by 33% (NNT=3). Larger than 5mm, increased by 34% (NNT=3)., Conclusion: Alpha-1 adrenergic blocker use is related with a greater incidence of expulsion of ureteral calculi, smaller or greater than 5mm, and fewer episodes of pain when compared to ibuprofen. However it is necessary larger samples to enhance the power analysis of the expulsion of ureteral calculi larger than 5mm and the episodes of pain., Patient Summary: This review analyzed the outcome of alpha adrenergic antagonist in children with ureteral calculi. We conclude that it is the best medicine for use, since it helps the expulsion of the stone.

Inductive Loop Detectors (ILDs) are the most commonly used sensors in traffic management systems. This paper shows that some spectral features extracted from the Fourier Transform (FT) of inductive signatures do not depend on the vehicle speed. Such a property is used to propose a novel method for vehicle classification based on only one signature acquired from a sensor single-loop, in contrast to standard methods using two sensor loops. Our proposal will be evaluated by means of real inductive signatures captured with our hardware prototype.

This study aimed at the isolation and characterization of metal(loid)-tolerant bacteria from the rhizosphere of Phragmites australis and Juncus effusus plants growing in two long-term contaminated sites in Northern Portugal. Site 1 had higher contamination than Site 3. Bacteria were isolated using metal(loid)-supplemented (Cd, Zn, and As) media. Isolates were grouped by random amplified polymorphic DNA and identified by 16S rRNA gene sequencing. Strains were also examined for their metal(loid) tolerance. The counts of metal(loid)-tolerant bacteria were higher in Site 1 and ranged between log 7.17 CFU g(-1) soil in As-containing medium and log 7.57 CFU g(-1) soil in Zn-containing medium, while counts at Site 3 varied between log 5.33 CFU g(-1) soil in Cd-containing medium and log 6.97 CFU g(-1) soil in As-containing medium. The composition of bacterial populations varied between locations. In Site 1, the classes Actinobacteria (36%) and Bacilli (24%) were well represented, while in Site 3 strains were mainly affiliated to classes Actinobacteria (35%), γ-Proteobacteria (35%), and β-Proteobacteria (12%). The order of metal(loid) toxicity for the isolated strains was Cd > As > Zn. Overall, 10 strains grew at 500 mg Cd L(-1) , 1000 mg Zn L(-1) , and 500 mg As L(-1) , being considered the most metal(loid)-tolerant bacteria. These strains belonged to genera Cupriavidus, Burkholderia, Novosphingobium, Sphingobacterium, Castellaniella, Mesorhizobium, Chryseobacterium, and Rhodococcus and were mainly retrieved from Site 1. The multiple metal(loid)-tolerant strains isolated in this study have potential to be used in bioremediation/phytoremediation., (© 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Therapeutic proteins and peptides demonstrate unique, peerless, pharmacological characteristics such as high specificity to receptors and superior biological mimicking of physiological mechanisms, resulting in a better therapeutic index compared to conventional chemical-derived drugs. However, proteins also present inherent bioavailability limitations. Thus, this paper proposes several effective tools to improve protein/peptide drugs stability, permeability and pharmacokinetics with special emphasis on oral polymeric films as oral delivery platforms. Indeed, oral films present inherent characteristics that can greatly enhance biological performance of proteins and peptides and patient compliance along with other advantages that are critically discussed in this review. A rational choice of excipients addressed in and manufacture processes are also focused. In addition, possible toxicity issues to be overtaken and critical analysis regarding current market tendencies respecting oral films and protein/peptides along with future prospects are disclosed., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Single phase hydroxyapatite (HAp) and biphasic material hydroxyapatite/β-tricalcium phosphate (HAp/β-TCP) were obtained from a marine source (Atlantic cod fish bones). Here we report a study on the biological properties of these materials, including cytotoxicity, bioactivity and haemocompatibility. Results showed that the materials are not cytotoxic, neither in their powder nor in pellet form; indeed growth of Saos-2 cells was comparable to that of commercial. The haemolysis rate was lower than 2%; hence the materials can be classified as non-haemolytic. Moreover, when immersed in Simulated Body Fluid (SBF), crystal formation was observed on the surface of both materials. The sintering behaviour of the samples was also studied; both powders showed very high sinterability (density higher than 95% of the theoretical value). Overall, these results confirm the suitability of these materials for biomedical applications., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Fluoxetine (FLX) is a chiral fluorinated pharmaceutical mainly indicated for treatment of depression and is one of the most distributed drugs. There is a clear evidence of environmental contamination with this drug. Aerobic granular sludge sequencing batch reactors constitute a promising technology for wastewater treatment; however the removal of carbon and nutrients can be affected by micropollutants. In this study, the fate and effect of FLX on reactor performance and on microbial population were investigated. FLX adsorption/desorption to the aerobic granules was observed. FLX shock loads (≤4μM) did not show a significant effect on the COD removal. Ammonium removal efficiency decreased in the beginning of first shock load, but after 20 days, ammonia oxidizing bacteria became adapted. The nitrite concentration in the effluent was practically null indicating that nitrite oxidizing bacteria was not inhibited, whereas, nitrate was accumulated in the effluent, indicating that denitrification was affected. Phosphate removal was affected at the beginning showing a gradual adaptation, and the effluent concentration was <0.04mM after 70 days. A shift in microbial community occurred probably due to FLX exposure, which induced adaptation/restructuration of the microbial population. This contributed to the robustness of the reactor, which was able to adapt to the FLX load., (Copyright © 2015 Elsevier B.V. All rights reserved.)

In this study, Pinus pinea seedlings mycorrhized with selected ectomycorrhizal fungi (ECMF), Pisolithus tinctorius and Suillus bellinii, were exposed to the herbicide benfluralin. Non-mycorrhized P. pinea seedlings and seedlings mycorrhized with ECMF were transferred to benfluralin-spiked soils at levels of 0.165, 1.65 and 16.5 mg kg(-1). Plant growth and the fungal role on plant antioxidant response were assessed. In the presence of benfluralin, higher plant growth was observed in mycorrhized plants compared to non-mycorrhized plants, but ECMF colonisation and nutrient uptake were affected by the herbicide. Benfluralin showed no effect on lipid peroxidation in P. pinea seedlings. However, seedlings mycorrhized with S. bellinii showed higher levels of lipid peroxidation when compared to non-mycorrhized ones, both in the presence and absence of benfluralin. The increase of lipid peroxidation could be related to seedling growth induced by the fungus and not to benfluralin toxicity. A similar trend was observed in seedlings mycorrhized with P. tinctorius when exposed to higher benfluralin concentrations, suggesting that the antioxidant response to benfluralin is related not only to fungus species, but also to the level of stress applied in the soil. The higher amount of superoxide dismutase activity in P. pinea seedlings tissues exposed to benfluralin could indicate a plant adaptative response to benfluralin toxicity. Catalase activity showed no increase with benfluralin exposure. Pre-established P. tinctorius mycorrhization conferred root protection and enhanced plant growth in benfluralin spiked soil, inferring that P. tinctorius - P. pinea association could advantageous for plant growth in soils contaminated with pesticides., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

In this study, we evaluated the phylogenetic diversity of culturable bacterial endophytes of Zea mays plants growing in an agricultural soil contaminated with Zn and Cd. Endophytic bacterial counts were determined in roots and shoots, and isolates were grouped by random amplified polymorphic DNA and identified by 16S ribosomal RNA (rRNA) gene sequencing. Endophytes were further characterized for the production of plant growth-promoting (PGP) substances, such as NH3, siderophores, indol-3-acetic acid (IAA), hydrogen cyanide and extracellular enzymes, and for the capacity to solubilize phosphate. The endophytes producing higher amounts of IAA were screened for their tolerance to Zn and Cd and used as bioinoculants for maize seedlings grown in the Zn/Cd-contaminated soil. The counts of endophytes varied between plant tissues, being higher in roots (6.48 log10 g(-1) fresh weight) when compared to shoots (5.77 log10 g(-1) fresh weight). Phylogenetic analysis showed that endophytes belong to three major groups: α-Proteobacteria (31 %), γ-Proteobacteria (26 %) and Actinobacteria (26 %). Pseudomonas, Agrobacterium, Variovorax and Curtobacterium were among the most represented genera. Endophytes were well-adapted to high Zn/Cd concentrations (up to 300 mg Cd l(-1) and 1,000 mg Zn l(-1)) and showed ability to produce several PGP traits. Strains Ochrobactrum haematophilum ZR 3-5, Acidovorax oryzae ZS 1-7, Frigoribacterium faeni ZS 3-5 and Pantoea allii ZS 3-6 increased root elongation and biomass of maize seedlings grown in soil contaminated with Cd and Zn. The endophytes isolated in this study have potential to be used in bioremediation/phytoremediation strategies.

Ectomycorrhizal fungi (ECMF) play an important role in forest ecosystems, often mitigating stress factors and increasing seedling performance. The aim of this study was to investigate the effects of a nursery inoculation on Pinus pinaster growth and on the fungal communities established when reforesting burned areas. Inoculated P. pinaster saplings showed 1.5-fold higher stem height than the non-inoculated controls after a 5 year growth period, suggesting that fungal inoculation could potentiate tree growth in the field. Ordination analysis revealed the presence of different ECMF communities on both plots. Among the nursery-inoculated fungi, Laccaria sp., Rhizopogon sp., Suillus bovinus and Pisolithus sp. were detected on inoculated Pinus saplings on both sampling periods, indicating that they persisted after field establishment. Other fungi were also detected in the inoculated plants. Phialocephala sp. was found on the first assessment, while Terfezia sp. was detected on both sampling periods. Laccaria sp. and Rhizopogon sp. were identified in the control saplings, belonging however to different species than those found in the inoculated plot. Inocybe sp., Thelephora sp. and Paxillus involutus were present on both sampling periods in the non-inoculated plots. The results suggest that ECMF inoculation at nursery stage can benefit plant growth after transplantation to a post-fire site and that the inoculated fungi can persist in the field. This approach has great potential as a biotechnological tool to aid in the reforestation of burned areas.

The interest for environmental fate assessment of chiral pharmaceuticals is increasing and enantioselective analytical methods are mandatory. This study presents an enantioselective analytical method for the quantification of seven pairs of enantiomers of pharmaceuticals and a pair of a metabolite. The selected chiral pharmaceuticals belong to three different therapeutic classes, namely selective serotonin reuptake inhibitors (venlafaxine, fluoxetine and its metabolite norfluoxetine), beta-blockers (alprenolol, bisoprolol, metoprolol, propranolol) and a beta2-adrenergic agonist (salbutamol). The analytical method was based on solid phase extraction followed by liquid chromatography tandem mass spectrometry with a triple quadrupole analyser. Briefly, Oasis MCX cartridges were used to preconcentrate 250 mL of water samples and the reconstituted extracts were analysed with a Chirobiotic V under reversed mode. The effluent of a laboratory-scale aerobic granular sludge sequencing batch reactor (AGS-SBR) was used to validate the method. Linearity (r(2)>0.99), selectivity and sensitivity were achieved in the range of 20-400 ngL(-1) for all enantiomers, except for norfluoxetine enantiomers which range covered 30-400 ngL(-1). The method detection limits were between 0.65 and 11.5 ngL(-1) and the method quantification limits were between 1.98 and 19.7 ngL(-1). The identity of all enantiomers was confirmed using two MS/MS transitions and its ion ratios, according to European Commission Decision 2002/657/EC. This method was successfully applied to evaluate effluents of wastewater treatment plants (WWTP) in Portugal. Venlafaxine and fluoxetine were quantified as non-racemic mixtures (enantiomeric fraction ≠ 0.5). The enantioselective validated method was able to monitor chiral pharmaceuticals in WWTP effluents and has potential to assess the enantioselective biodegradation in bioreactors. Further application in environmental matrices as surface and estuarine waters can be exploited., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Fluoxetine (FLX) is a chiral fluorinated pharmaceutical indicated mainly for the treatment of depression and is one of the most dispensed drugs in the world. There is clear evidence of environmental contamination with this drug and its active metabolite norfluoxetine (NFLX). In this study the enantioselective biodegradation of racemic FLX and of its enantiomers by Labrys portucalensis strain F11 was assessed. When 2μM of racemic FLX was supplemented as sole carbon source, complete removal of both enantiomers, with stoichiometric liberation of fluoride, was achieved in 30d. For racemic FLX concentration of 4 and 9μM, partial degradation of the enantiomers was obtained. In the presence of acetate as an additional carbon source, at 4, 9 and 21μM of racemic FLX and at 25μM of racemic FLX, (S)-FLX or (R)-FLX, complete degradation of the two enantiomers occurred. At higher concentrations of 45 and 89μM of racemic FLX, partial degradation was achieved. Preferential degradation of the (R)-enantiomer was observed in all experiments. To our knowledge, this is the first time that enantioselective biodegradation of FLX by a single bacterium is reported., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

The release of fluorinated organic compounds from fire retardants or agrochemical products may have a significant negative effect on soil ecosystems. In this study, the ability of Pisolithus tinctorius to tolerate and degrade mono-fluorophenols (FP) was assessed. In vitro studies showed fungal growth in the presence of 0.45mM of 2-FP and 3-FP, but not in the presence of 4-FP. P. tinctorius was able to degrade up to 79% and 92% of 1mM 2-FP and 3-FP, respectively, in glucose supplemented liquid medium, suggesting that 2- and 3-FP degradation occurred in co-metabolism with glucose consumption. 3-Fluorocatechol (FC) and 4-FC were identified as metabolic intermediates using HPLC and LC-MS. Liberation of fluoride was not detected suggesting that a fluorinated dead-end product was formed. In extracts of cells collected at the end of cultures supplemented with the mono-FPs, a metabolic intermediate compatible with a mass corresponding to a fluoromuconate compound, according to LC-MS data, was recovered. The results further suggest that ectomycorrhizal fungi may be able to degrade mono-FP in pure culture while using glucose as a carbon source, through a similar pathway as that found in bacteria. To our knowledge, this is the first time that degradation of mono-FPs by an ectomycorrhizal fungus is reported., (Copyright © 2014 Elsevier Ltd. All rights reserved.)