Evaluation of deep eutectic solvents in the synthesis of molecularly imprinted fibers for the solid-phase microextraction of triazines in soil samples.

Nowadays, molecularly imprinted polymers (MIPs) are well established and are considered excellent materials for performing selective extractions. However, with the progressive implementation of the principles of green chemistry, it is necessary to find greener alternatives for both the synthesis and further use of MIPs in sample preparation. Accordingly, in the present work, different deep eutectic solvents (DES, both hydrophilic and hydrophobic), as an alternative to conventional organic solvents (i.e., toluene), were evaluated as porogens for the synthesis of imprinted fibers (monoliths), using fused silica capillaries as molds, for solid-phase microextraction (SPME). From this study, the polymer prepared with propazine (dummy template), methacrylic acid (monomer), ethylene glycol dimethacrylate (cross-linker), and a formic acid:L-menthol (1:1) DES (porogen) showed the best performance for selective rebinding of triazines. After optimization of the different variables involved in SPME, the new imprinted fibers were successfully applied to the extraction of target analytes (desisopropylatrazine, desethylatrazine, simazine, and atrazine) from soil sample extracts, providing relative recoveries ranging from 75.7 to 120.1%, reaching limits of detection within the range of 6.2-15.7 ng g-1, depending upon the analyte.

Sardines at a junction: Seascape genomics reveals ecological and oceanographic drivers of variation in the NW Mediterranean Sea.

By evaluating genetic variation across the entire genome, one can address existing questions in a novel way while raising new ones. The latter includes how different local environments influence adaptive and neutral genomic variation within and among populations, providing insights into local adaptation of natural populations and their responses to global change. Here, under a seascape genomic approach, ddRAD data of 4609 single nucleotide polymorphisms (SNPs) from 398 sardines (Sardina pilchardus) collected in 11 Mediterranean and one Atlantic site were generated. These were used along with oceanographic and ecological information to detect signals of adaptive divergence with gene flow across environmental gradients. The studied sardines constitute two clusters (FST  = 0.07), a pattern attributed to outlier loci, highlighting putative local adaptation. The trend in the number of days with sea surface temperature above 19°C, a critical threshold for successful sardine spawning, was crucial at all levels of population structuring with implications on the species' key biological processes. Outliers link candidate SNPs to the region's environmental heterogeneity. Our findings provide evidence for a dynamic equilibrium in which population structure is maintained by physical and ecological factors under the opposing influences of migration and selection. This dynamic in a natural system warrants continuous monitoring under a seascape genomic approach that might benefit from a temporal and more detailed spatial dimension. Our results may contribute to complementary studies aimed at providing deeper insights into the mechanistic processes underlying population structuring. Those are key to understanding and predicting future changes and responses of this highly exploited species in the face of climate change.

Recent advances and future trends in molecularly imprinted polymers-based sample preparation.

Molecular imprinting technology is a well-established technique for the obtainment of tailor-made polymers, so-called molecularly imprinted polymers, with a predetermined selectivity towards a target analyte or structurally related compounds. Accordingly, molecularly imprinted polymers are considered excellent materials for sample preparation providing unprecedented selectivity to analytical methods. However, the use of molecularly imprinted polymers in sample preparation still presents some shortcomings derived from the synthesis procedure itself limiting its general applicability. In this regard, molecularly imprinted polymers use to display binding sites heterogeneity and slow diffusion mass transfer of analytes to the imprinted sites affecting their overall performance. Besides, the performance of molecularly imprinted polymers in organic solvents is excellent, but their selective binding ability in aqueous media is considerably reduced. Accordingly, the present review pretends to provide an updated overview of the recent advances and trends of molecularly imprinted polymers-based extraction, focusing on those strategies proposed for the improvement of mass transfer and selective recognition in aqueous media. Besides, with the progressive implementation of Green Chemistry principles, the different steps and strategies for the preparation of molecularly imprinted polymers are reviewed from a green perspective.

Pain Behaviors Analyzed by Videorecording in Brain-Injured Patients Admitted to the Intensive Care Unit.

AIM: To describe and classify pain behaviors (facial and body) in brain-injured patients with a low level of consciousness before, during, and after the performance of painful and non-painful care procedures. METHODS: Facial behaviors and body movements in brain-injured patients were videotaped at rest, during the application of three care procedures (two painful and one non-painful), and 15 minutes after completion of these procedures. Each video recording was evaluated by expert evaluators blinded to each other. For each of the behaviors observed, all possible combinations between the three procedures and/or time were compared using the McNemar test. Effect size was measured by the difference in proportions using the Wilson score 95% confidence intervals. RESULTS: Twenty-seven patients were included. The mean (standard deviation) Glasgow Coma Score was 5.4 (1.9). A total of 33 behaviors (29 active, four neutral) were registered. Expression of behaviors was more common during the painful procedures compared with the other time points (non-painful procedures, baseline, and final evaluation). Inter-evaluator agreement was substantial (Kappa index >0.7) in more than 50% of the observed behaviors. CONCLUSIONS: In this study involving brain-injured patients with a low level of consciousness, facial, body, and ventilation-related behaviors were more common during painful procedures. Agreement between evaluators to detect the presence or absence of these behaviors was substantial. These findings underscore the need to develop pain assessment measures specific to this patient population.

Resilience dynamics and productivity-driven shifts in the marine communities of the Western Mediterranean Sea.

Ecological resilience has become a conceptual cornerstone bridging ecological processes to conservation needs. Global change is increasingly associated with local changes in environmental conditions that can cause abrupt ecosystem reorganizations attending to system-specific resilience fluctuations with time (i.e. resilience dynamics). Here we assess resilience dynamics associated with climate-driven ecosystems transitions, expressed as changes in the relevant contribution of species with different life-history strategies, in two benthopelagic systems. We analysed data from 1994 to 2019 coming from a scientific bottom trawl survey in two environmentally contrasting ecosystems in the Western Mediterranean Sea-Northern Spain and Alboran Sea. Benthopelagic species were categorized according to their life-history strategies (opportunistic, periodic and equilibrium), ecosystem functions and habitats. We implemented an Integrated Resilience Assessment (IRA) to elucidate the response mechanism of the studied ecosystems to several candidate environmental stressors and quantify the ecosystems' resilience. We demonstrate that both ecosystems responded discontinuously to changes in chlorophyll-a concentration more than any other stressor. The response in Northern Spain indicated a more overarching regime shift than in the Alboran Sea. Opportunistic fish were unfavoured in both ecosystems in the recent periods, while invertebrate species of short life cycle were generally favoured, particularly benthic species in the Alboran Sea. The study illustrates that the resilience dynamics of the two ecosystems were mostly associated with fluctuating productivity, but subtle and long-term effects from sea warming and fishing reduction were also discernible. Such dynamics are typical of systems with wide environmental gradient such as the Northern Spain, as well as systems with highly hydrodynamic and of biogeographical complexity such as the Alboran Sea. We stress that management should become more adaptive by utilizing the knowledge on the systems' productivity thresholds and underlying shifts to help anticipate both short-term/less predictable events and long-term/expected effects of climate change.

Green molecularly imprinted polymers for sustainable sample preparation.

The use of molecularly imprinted polymers in sample preparation as selective sorbent materials has received great attention during the last years leading to analytical methods with unprecedented selectivity. However, with the progressive implementation of Green Analytical Chemistry principles, it is necessary to critically review the greenness of synthesis and further use of molecularly imprinted polymers in sample preparation. Accordingly, in the present review, the different steps and strategies for the preparation of molecularly imprinted polymers, the used reagents, as well as their incorporation to microextraction techniques are reviewed from a green perspective and recent alternatives to make the use of molecularly imprinted polymers more sustainable are provided.

Evaluation of 2-hydroxyethyl methacrylate as comonomer in the preparation of water-compatible molecularly imprinted polymers for triazinic herbicides.

In this work, the preparation and evaluation of water-compatible molecularly imprinted polymers for triazines using 2-hydroxyethyl methacrylate and methacrylic acid as comonomers is described. Four sets of molecularly imprinted and non-imprinted polymers for propazine were prepared at varying monomer molar ratios (from 4:0 to 1:3), and evaluated for the recognition of several triazines directly in aqueous media. The evaluation was performed by loading 1 mL of an aqueous solution containing 500 ng of each selected triazine, washing with 500 µL of acetonitrile, and eluting with 500 µL of methanol followed by 2 × 500 µL of a solution of methanol containing 10% of acetic acid. Final determinations were performed by high-performance liquid chromatography-ultraviolet detection. Improvement in molecular recognition of triazines in water was obtained on those molecularly imprinted polymers incorporating 2-hydroxyethyl methacrylate in 3:1 or 2:2 molar ratios, being the former selected as optimum providing recoveries for propazine up to 80%. A molecularly imprinted solid-phase extraction protocol was developed to ensure that triazines-selective recognition takes place inside selective binding sites in pure water media. Finally, the developed method was successfully applied to the determination of the selected triazines in environmental waters providing limits of detection from 0.16 and the 0.5 µg/L concentration range.

Surface modified-magnetic nanoparticles by molecular imprinting for the dispersive solid-phase extraction of triazines from environmental waters.

Magnetic nanoparticles have been surface modified by molecular imprinting and evaluated as selective sorbents for the extraction of triazines from environmental waters. The use of propazine as template allowed us to synthesize a selective material able to simultaneously recognize and selective extract not only the template but also several other herbicides of the same family. A magnetic molecularly imprinted-based dispersive solid-phase extraction procedure was developed and fully optimized. Magnetic molecularly imprinted polymer particles can be easily collected and separated from liquid solvents and samples with the help of an external magnetic field, avoiding in that way any centrifugation or filtration steps, which represents a remarkable advantage over traditional procedures. Under optimum conditions, selective extraction of several triazines (cyanazine, simazine, atrazine, propazine, and terbutylazine) from environmental water samples was performed prior to final determination by high-performance liquid chromatography with diode-array detection. Recoveries for the studied triazines were within the range of 75.2-94.1%, with relative standard deviations lower than 11.3% (n = 3). The limits of detection were within 0.16-0.51 µg/L, depending upon the triazine and the type of sample analyzed.

Application of the Behavioural Indicators of Pain Scale in patients with traumatic brain injury.

AIM: To develop and psychometrically test the Behavioural Indicators of Pain Scale (ESCID) in patients with traumatic brain injury (TBI). DESIGN: A prospective observational study to test the psychometric properties of the Behavioural Indicators of Pain Scale in patients with TBI. METHOD: A convenience sample of patients with TBI, who were non-communicative and using invasive mechanical ventilation was selected. Pain was evaluated by two observers who were blinded from each other. Assessments were performed at baseline via the performance of a painful procedure (aspiration of secretions) and a non-painful procedure (rubbing with a gauze). Assessments were repeated after application of procedures on days 1 and 6 of hospitalization in an intensive care unit. Data were collected between January-December 2016. RESULTS: About 134 patients were included in the study. Of these, 76.1% were men. The mean age of participants was 45.2 (SD 17.5) years. The pain score significantly increased during the painful procedure when compared with the baseline measure and non-painful procedure (p < .001). Patients displayed a greater number of pain-indicating behaviours during the painful procedure on day 6, compared with day 1 (p < .05). This finding coincided with a reduced level of sedation and a greater level of consciousness. CONCLUSION: The ESCID scale detects pain behaviours and discriminates among the different types of stimulation in patients with brain injury, who are uncommunicative and with mechanical ventilation, with good reliability. The ability for patients with brain injury to express behaviours is limited because of the low level of consciousness and the deep level of sedation. IMPACT: This research will have an impact on the practice of pain assessment in patients with brain injury, representing a first step to adapt the content of the ESCID.

Molecularly imprinted core-shell magnetic nanoparticles for selective extraction of triazines in soils.

In this work, a propazine-imprinted polymer was synthesized on the surface of modified magnetic nanoparticles to be used in the solid-phase extraction of triazines in soil samples. The effect of different solvents on the selective extraction of target analytes was assessed to establish the optimum rebinding conditions. The obtained magnetic molecularly imprinted particles exhibited high selectivity for triazines and were easily collected and separated by an external magnetic field without additional centrifugation or filtration steps. Under optimum conditions, a magnetic molecularly imprinted solid-phase extraction method was developed allowing the extraction of several triazines (desisopropylatrazine, desethylatrazine, simazine, atrazine, and propazine) from soil samples and their subsequent final determination by high-performance liquid chromatography with diode-array detection. Recoveries for the triazines studied were within the range 5.4% to 40.6%, with relative standard deviations lower than 7.0% (n = 3). The detection limits were within 0.1 to 3 ng g-1 , depending upon the triazine and the type of soil used.

Molecularly imprinted magnetic nanoparticles for the micro solid-phase extraction of thiabendazole from citrus samples.

The preparation of molecularly imprinted core-shell magnetic nanoparticles and their subsequent use in the solid-phase extraction of thiabendazole from citrus sample extracts is described. Molecularly imprinted core-shell magnetic nanoparticles were prepared by the precipitation copolymerization of the imprinting polymerization mixture on the surface of vinyl-modified silica magnetic nanoparticles and were characterized by Fourier transform infrared spectroscopy and scanning electron microscopy. The obtained molecularly imprinted core-shell magnetic nanoparticles exhibited a high selectivity for thiabendazole and were easily collected and separated by an external magnetic field without additional centrifugation or filtration steps. Under optimum conditions, a magnetic molecularly imprinted solid-phase extraction method was developed allowing the extraction of thiabendazole from citrus sample extracts and final determination by high-performance liquid chromatography with fluorescence detection. The detection limit was 0.2 mg/kg, far lower than the maximum residue limit established within the European Union for thiabendazole in citrus samples.

Preparation of molecularly imprinted polymeric fibers using a single bifunctional monomer for the solid-phase microextraction of parabens from environmental solid samples.

In this study, molecularly imprinted polymer fibers for solid-phase microextraction have been prepared with a single bifunctional monomer, N,O-bismethacryloyl ethanolamine using the so-called "one monomer molecularly imprinted polymers" method, replacing the conventional combination of functional monomer and cross-linker to form high fidelity binding sites. For comparison, imprinted fibers were prepared following the conventional approach based on ethylene glycol dimethacrylate as cross-linker and methacrylic acid as monomer. The recognition performance of the new fibers was evaluated in the solid-phase microextraction of parabens, and from this study it was concluded that they provided superior performance over conventionally formulated fibers. Ultimately, real-world environmental testing on spiked solid samples was successful by the molecularly imprinted solid-phase microextraction of samples, and the relative recoveries obtained at enrichment levels of 10 ng/g of parabens were within 78-109% for soil and 83-109% for sediments with a relative standard deviation <15% (n = 3).

Evaluation of electrochemically synthesized sulfadimethoxine-imprinted polymer for solid-phase microextraction of sulfonamides.

Solid-phase microextraction (SPME) is widely used in analytical laboratories for the analysis of organic compounds, thanks to its simplicity and versatility. In the present work, the synthesis and evaluation of imprinted films for SPME by electropolymerisation of pyrrole alone or in the presence of ethylene glycol dimethacrylate is proposed. Sulfadimethoxine (SDM), a sulfonamide antibiotic, was used as template molecule. Initially, a molecularly imprinted polymer film was prepared by electropolymerisation of pyrrole onto a platinum foil, using SDM as template. The SDM template was removed by overoxidation. The behaviour of SDM on imprinted and non-imprinted polymers was investigated by differential pulse voltammetry, and a clear imprinting effect was observed, which was confirmed by rebinding experiments using both conventional and electrochemically enhanced-SPME. However, in general, the extraction efficiency was rather low (<6%) and unspecific interactions are too high. Attempts to increase extraction efficiency were unsuccessful, but the incorporation of ethylene glycol dimethacrylate to the films reduced unspecific interactions to a certain extent.

Spatial patterns of ß-diversity under cumulative pressures in the Western Mediterranean Sea.

Understanding the spatial dynamics of biodiversity is an essential issue in marine ecology and requires combining information at local and regional scales. ß-diversity is an important measure of biodiversity that informs on the differences in community composition between sites and, thus, in the species turnover in the community structure. In this study, we analysed and predicted the spatial patterns of ß-diversity for fishes, invertebrates and the demersal assemblage along the Iberian Mediterranean coast. We used Bayesian Bootstrap Generalized Dissimilarity Models (BBGDMs) to study the effects of environment and human pressures on the ß-diversity of invertebrate, fishes and the entire demersal assemblage from 1994 to 2015 using different time windows to account for temporal variability. Then, we used these relationships to predict the spatial patterns of ß-diversity in the whole Iberian Mediterranean coast. Our results highlighted that the regional spatial patterns of ß-diversity were best described by bathymetry and a cumulative index of coastal impacts. We identified specific regions with the highest ß-diversity in the study area, which were complementary to hotspots of species richness and presented different degree of overlapping with existent marine protected areas. Overall, our study illustrates that by modelling spatial turnover using ß-diversity we can better understand and predict spatial variation of biodiversity and the effects of particular variables, providing relevant information to end-users and policy makers for designing specific spatial conservation and management strategies.

Understanding the response of the Western Mediterranean cephalopods to environment and fishing in a context of alleged winners of change.

Increasing impacts of both fisheries and climate change have resulted in shifts in the structure and functioning of marine communities. One recurrent observation is the rise of cephalopods as fish recede. This is generally attributed to the removal of main predators and competitors by fishing, while mechanistic evidence is still lacking. In addition, climate change may influence cephalopods due to their high environmental sensitivity. We aim to unveil the effects of different anthropogenic and environmental drivers at different scales focusing on the cephalopod community of the Western Mediterranean Sea. We investigate several ecological indicators offering a wide range of information about their ecology, and statistically relating them with environmental, biotic and fisheries drivers. Our results highlight non-linear changes of indicators along with spatial differences in their responses. Overall, the environment drivers have greater effects than biotic and local human impacts with contrasting effects of temperature across the geographic gradient. We conclude that cephalopods may be impacted by climate change in the future while not necessary through positive warming influence, which should make us cautious when referring to them as generalized winners of current changes.

AGREEMIP: The Analytical Greenness Assessment Tool for Molecularly Imprinted Polymers Synthesis.

Molecular imprinting technology is well established in areas where a high selectivity is required, such as catalysis, sensing, and separations/sample preparation. However, according to the Principles of Green Chemistry, it is evident that the various steps required to obtain molecularly imprinted polymers (MIPs) are far from ideal. In this regard, greener alternatives to the synthesis of MIPs have been proposed in recent years. However, although it is intuitively possible to design new green MIPs, it would be desirable to have a quantitative measure of the environmental impact of the changes introduced for their synthesis. In this regard, this work proposes, for the first time, a metric tool and software (termed AGREEMIP) to assess and compare the greenness of MIP synthesis procedures. AGREEMIP is based on 12 assessment criteria that correspond to the greenness of different reaction mixture constituents, energy requirements, and the details of MIP synthesis procedures. The input data of the 12 criteria are transformed into individual scores on a 0-1 scale that in turn produce an overall score through the calculation of the weighted average. The assessment can be performed using user-friendly open-source software, freely downloadable from mostwiedzy.pl/agreemip. The assessment result is an easily interpretable pictogram and visually appealing, showing the performance in each of the criteria, the criteria weights, and overall performance in terms of greenness. The application of AGREEMIP is presented with selected case studies that show good discrimination power in the greenness assessment of MIP synthesis pathways.

The trade-off between condition and growth shapes juveniles' survival of harvested demersal fish of the Mediterranean sea.

Fish body condition and growth are two interrelated traits closely associated with species life history and fitness, whose trade-off can ultimately impact population dynamics albeit seldom empirically demonstrated. They can intricately affect survival rates, which are particularly relevant for species under exploitation. Using individual spatiotemporal information in Northwestern Mediterranean, we document for the first time the existence of a trade-off between condition and growth in regulating survival dynamics in two important fish species for the Mediterranean fisheries that are characterized by contrasting life histories. For the European hake (Merluccius merluccius), a benthopelagic species, juveniles' body condition was detected to be positively linked to survival and negatively associated with the growth of this age group. For the red mullet (Mullus barbatus), the same pattern was observed for young adults. We also show that the observed patterns on a regional level have a clear spatial dependence as we found that observed body condition over a local scale had a broad effect on the population dynamics of the whole region, with the Ebro delta area emerging as the demographic engine of the two species. We discuss our results in the context of fisheries management and underline the importance of improving current stock assessment models and spatially based fishery management towards incorporating body condition and growth due to their influence on important parameters such as survival.

Molecularly imprinted stir bars for selective extraction of thiabendazole in citrus samples.

Stir-bar sorptive extraction is based on the partitioning of target analytes between the sample (mostly aqueous-based liquid samples) and a stationary phase-coated magnetic stir bar. Until now, only PDMS-coated stir bars are commercially available, restricting the range of applications to the non-selective extraction of hydrophobic compounds due to the apolar character of PDMS. In this work, a novel stir bar coated with molecularly imprinted polymer as selective extraction phase for sorptive extraction of thiabendazole (TBZ) was developed. Two different procedures, based on physical or chemical coating, were assessed for the preparation of molecularly imprinted stir bars. Under optimum conditions, recoveries achieved both in imprinted and non-imprinted polymer stir bars obtained by physical coating were very low, whereas TBZ was favourably retained by imprinted over non-imprinted polymer stir bars obtained by chemical coating and thus the latter approach was used in further studies. Different parameters affecting both stir-bars preparation (i.e. cross-linker, porogen, polymerization time) and the subsequent selective extraction of TBZ (i.e. washing, loading and elution solvents, extraction time) were properly optimized. The molecularly imprinted coated stir bars were applied to the extraction of TBZ from citrus samples (orange, lemon and citrus juices) allowing its final determination at concentrations levels according to current regulations.

Organophosphate ester plasticizers in edible fish from the Mediterranean Sea: Marine pollution and human exposure.

Concentrations of organophosphate esters (OPEs) plasticizers were analysed in the present study. Fifty-five fish samples belonging to three highly commercial species, European sardine (Sardina pilchardus), European anchovy (Engraulis encrasicolus), and European hake (Merluccius merluccius), were taken from the Western Mediterranean Sea. OPEs were detected in all individuals, except for two hake samples, with concentrations between 0.38 and 73.4 ng/g wet weight (ww). Sardines presented the highest mean value with 20.5 ± 20.1 ng/g ww, followed by anchovies with 14.1 ± 8.91 ng/g ww and hake with 2.48 ± 1.76 ng/g ww. The lowest OPE concentrations found in hake, which is a partial predator of anchovy and sardine, and the higher δ15N values (as a proxy of trophic position), may indicate the absence of OPEs biomagnification. Eleven out of thirteen tested OPEs compounds were detected, being diphenyl cresyl phosphate (DCP) one of the most frequently detected in all the species. The highest concentration values were obtained for tris(1,3-dichloro-2-propyl) phosphate (TDClPP), trihexyl phosphate (THP), and tris(2-butoxyethyl) phosphate (TBOEP), for sardines, anchovies, and hakes, respectively. The human health risk associated with the consumption of these fish species showing that their individual consumption would not pose a considerable threat to public health regarding OPE intake.

Determination of polypeptide antibiotics in animal tissues using liquid chromatography tandem mass spectrometry based on in-line molecularly imprinted solid-phase extraction.

Effective purification and enrichment of polypeptide antibiotics in animal tissues is always a challenge, due to the co-extraction of other endogenous peptides which usually interfere their final determination. In this study, a molecularly imprinted column was prepared by packing polymyxin E-imprinted particles into a 100 mm × 4.6 mm i.d. HPLC column. The as-prepared imprinted columns were able to tolerate 100% aqueous phase and exhibited good stability and high column efficiency. Polypeptides antibiotics with similar molecular size or spatial structure to polymyxin E were well retained by the imprinted column, suggesting class selectivity. After optimization of mobile phase conditions of imprinted column, polypeptide antibiotics in animal tissue extracts were enriched and cleaned up by in-line molecularly imprinted solid-phase extraction, allowing the screening of target analytes in complex samples at low concentration levels by UV detection. Eluate fraction from the imprinted column was collected, and further dried and re-dissolved with methanol-0.5% formic acid aqueous solution (80:20, v/v) for final LC-MS/MS analysis. Analysis was accomplished using multiple reaction monitoring (MRM) in positive electrospray ionization mode and analytes quantified using the matrix-matched external calibration curves. The results showed high correlation coefficients for target analytes in the linear range of 2 ∼ 200 µg kg-1. At four different concentration levels (limit of quantification, 50, 100 and 200 µg kg-1), recoveries of four polypeptide antibiotics in swine, cattle and chicken muscles ranged from 66.7 to 94.5% with relative standard deviations lower than 16.0%. The limits of detection (LOD) were 2.0 ∼ 4.0 µg/kg, depending upon the analyte and sample. Compared with a conventional pretreatment method, the imprinted column was able to remove more impurities and to significantly reduce matrix effects, allowing the accurate analysis of polypeptide antibiotics.