Assessing the Effect of Surface Coating on the Stability, Degradation, Toxicity and Cell Endocytosis/Exocytosis of Upconverting Nanoparticles.

Lanthanide-doped up-converting nanoparticles (UCNPs) have emerged as promising biomedical tools in recent years. Most research efforts were devoted to the synthesis of inorganic cores with the optimal physicochemical properties. However, the careful design of UCNPs with the adequate surface coating to optimize their biological performance still remains a significant challenge. Here, we propose the functionalization of UCNPs with four distinct types of surface coatings, which were compared in terms of the provided colloidal stability and resistance to degradation in different biological-relevant media, including commonly avoided analysis in acidic lysosomal-mimicking fluids. Moreover, the influence of the type of particle surface coating on cell cytotoxicity and endocytosis/exocytosis was also evaluated. The obtained results demonstrated that the functionalization of UCNPs with poly(isobutylene-alt-maleic anhydride) grafted with dodecylamine (PMA-g-dodecyl) constitutes an outstanding strategy for their subsequent biomedical application, whereas poly(ethylene glycol) (PEG) coating, although suitable for colloidal stability purposes, hinders extensive cell internalization. Conversely, surface coating with small ligand were found not to be suitable, leading to large degradation degrees of UCNPs. The analysis of particle' behavior in different biological media and in vitro conditions here performed pretends to help researchers to improve the design and implementation of UCNPs as theranostic nanotools.

Evaluation of an Oral Fluid Collection Device and a Solid-Phase Extraction Method for the Determination of Coca Leaf Alkaloids by Gas Chromatography-Mass Spectrometry.

Some South American countries have ancient traditions that may pose legal problems, such as the consumption of coca leaves, as this can provide positive results for cocaine use after the analysis of biological samples. For this reason, it is necessary to find specific markers that help differentiate legal from illegal consumption, such as tropacocaine, cinnamoylcocaine, and especially hygrine and cuscohygrine. In this work, two techniques for collecting biological samples are compared: the Quantisal® Oral Fluid collection device and passive drooling. Once the samples were collected, they were subjected to solid-phase extraction for subsequent injection into GC-MS. Different validation parameters included in international guides have been studied to evaluate whether the proposed method is valid for the defined purpose, placing special emphasis on the study of the matrix effect and little value on GC-MS analyses. With respect to this parameter, an increase in the signal was found for CUS and t-CIN, but it was not significant for the rest of the substances studied. The recoveries have varied significantly depending on the way of working, being higher when working with standardized areas. After carrying out work with the oral fluid samples collected from laboratory volunteers, the method was applied to two real samples. The results obtained support the need for further research to overcome certain limitations presented by the device.

A new series of acylhydrazones derived from metribuzin with modulated herbicidal activity.

This paper reports the preparation and herbicidal evaluation of a small library of acylhydrazones based on the synthetic herbicide metribuzin. The hydrazone linkage easily obtained by reaction of metribuzin with aliphatic and aromatic aldehydes, masks efficiently the exocyclic amino group, thereby altering significantly H-bonding with the receptor and increasing the lipophilicity relative to the parent herbicide. The structures of all compounds, including key stereochemical issues on conformation and E/Z configuration around the C[bond, double bond]N bond were thoroughly elucidated by spectroscopic methods, and unambiguously corroborated by X-ray diffraction analysis. The herbicidal assays using an aliphatic and an aromatic acylhydrazone were performed on tomato and rapeseed plants grown in greenhouse. Our results demonstrate, regardless of rate application, that such acylhydrazone formulations do not alter the selectivity of metribuzin. Moreover, the herbicide activity was even higher in the alkyl derivative than that achieved by commercial metribuzin, thus suggesting that this substance can be applied with no need of combination with chemical coadjuvants, unlike most formulations of commercially available herbicides. Therefore, the study shows the promising effect of chemical derivatization of a common herbicide as metribuzin, to improve the herbicide activity without compromising selectivity, and allowing the farmers its use in crop protection safely and effectively.

Ultrasound assisted membrane-assisted solvent extraction for the simultaneous assessment of some drugs involved in drug-facilitated sexual assaults by liquid chromatography-tandem mass spectrometry.

A simple and highly efficient ultrasound assisted membrane-assisted solvent extraction (MASE) pre-treatment method for urine has been developed and validated for the simultaneous determination of twenty-two drugs involved in drug-facilitated sexual assaults (DFSAs) by liquid chromatography-tandem mass spectrometry. MASE was performed with 4.0 mL of urine (pH adjusted at 12), 400 µL of hexane as an organic solvent inside the polypropylene membrane, and ultrasonication (45 kHz, 120 W) for 10 min. A pre-concentration factor of 40 was achieved after evaporation (N2 stream) and re-dissolution in 100 µL of methanol. Analytes were separated using a Zorbax Eclipse Plus C18 column under gradient elution with aqueous 10 mM NH4HCO3 (pH 8.0) and methanol as mobile phases. Matrix-matched calibrations allowed the assessment of DFSA drugs of quite different octanol-water partition coefficients (Ko/w), from 1.32 101 for pregabalin to 2.45 105 for clomipramine (Log P values from 1.12 (pregabalin) to 5.39 (clomipramine)). The limit of detection (LOD) was between 0.0075 to 0.37 µg L-1, with analytical recoveries ranging from 73 to 103%, and relative standard deviations (RSDs) within the 2-20% range. The applicability of the method was demonstrated after analysing urine samples under forensic investigation.

Acute Aquatic Toxicity to Zebrafish and Bioaccumulation in Marine Mussels of Antimony Tin Oxide Nanoparticles.

Antimony tin oxide (Sb2O5/SnO2) is effective in the absorption of infrared radiation for applications, such as skylights. As a nanoparticle (NP), it can be incorporated into films or sheets providing infrared radiation attenuation while allowing for a transparent final product. The acute toxicity exerted by commercial Sb2O5/SnO2 (ATO) NPs was studied in adults and embryos of zebrafish (Danio rerio). Our results suggest that these NPs do not induce an acute toxicity in zebrafish, either adults or embryos. However, some sub-lethal parameters were altered: heart rate and spontaneous movements. Finally, the possible bioaccumulation of these NPs in the aquacultured marine mussel Mytilus sp. was studied. A quantitative analysis was performed using single particle inductively coupled plasma mass spectrometry (sp-ICP-MS). The results indicated that, despite being scarce (2.31 × 106 ± 9.05 × 105 NPs/g), there is some accumulation of the ATO NPs in the mussel. In conclusion, commercial ATO NPs seem to be quite innocuous to aquatic organisms; however, the fact that some of the developmental parameters in zebrafish embryos are altered should be considered for further investigation. More in-depth analysis of these NPs transformations in the digestive tract of humans is needed to assess whether their accumulation in mussels presents an actual risk to humans.

Bioaccumulation of titanium dioxide nanoparticles in green (Ulva sp.) and red (Palmaria palmata) seaweed.

A bioaccumulation study in red (Palmaria palmata) and green (Ulva sp.) seaweed has been carried out after exposure to different concentrations of citrate-coated titanium dioxide nanoparticles (5 and 25 nm) for 28 days. The concentration of total titanium and the number and size of accumulated nanoparticles in the seaweeds has been determined throughout the study by inductively coupled plasma mass spectrometry (ICP-MS) and single particle-ICP-MS (SP-ICP-MS), respectively. Ammonia was used as a reaction gas to minimize the effect of the interferences in the 48Ti determination by ICP-MS. Titanium concentrations measured in Ulva sp. were higher than those found in Palmaria palmata for the same exposure conditions. The maximum concentration of titanium (61.96 ± 15.49 µg g-1) was found in Ulva sp. after 28 days of exposure to 1.0 mg L-1 of 5 nm TiO2NPs. The concentration and sizes of TiO2NPs determined by SP-ICP-MS in alkaline seaweed extracts were similar for both seaweeds exposed to 5 and 25 nm TiO2NPs, which indicates that probably the element is accumulated in Ulva sp. mainly as ionic titanium or nanoparticles smaller than the limit of detection in size (27 nm). The implementation of TiO2NPs in Ulva sp. was confirmed by electron microscopy (TEM/STEM) in combination with energy dispersive X-Ray analysis (EDX).

Quantitative titanium imaging in fish tissues exposed to titanium dioxide nanoparticles by laser ablation-inductively coupled plasma-mass spectrometry.

Imaging studies by laser ablation-inductively coupled plasma mass spectrometry have been successfully developed to obtain qualitative and quantitative information on the presence/distribution of titanium (ionic titanium and/or titanium dioxide nanoparticles) in sea bream tissues (kidney, liver, and muscle) after exposure assays with 45-nm citrate-coated titanium dioxide nanoparticles. Laboratory-produced gelatine standards containing ionic titanium were used as a calibration strategy for obtaining laser ablation-based images using quantitative (titanium concentrations) data. The best calibration strategy consisted of using gelatine-based titanium standards (from 0.1 to 2.0 µg g-1) by placing 5.0-µL drops of the liquid gelatine standards onto microscope glass sample holders. After air drying at room temperature good homogeneity of the placed drops was obtained, which led to good repeatability of measurements (calibration slope of 4.21 × 104 ± 0.39 × 104, n = 3) and good linearity (coefficient of determination higher than 0.990). Under the optimised conditions, a limit of detection of 0.087 µg g-1 titanium was assessed. This strategy allowed to locate prominent areas of titanium in the tissues as well as to quantify the bioaccumulated titanium and a better understanding of titanium dioxide nanoparticle spatial distribution in sea bream tissues.

Effects of fresh and field-aged holm-oak biochar on As, Cd and Pb bioaccumulation in different rice growing environments.

Arsenic, Cd, and Pb environmental fate is influenced when the traditional permanent flooding rice production systems are replaced by water-saving and soil conservation practices, urging for additional strategies that avoid their bioaccumulation in rice grain. The aim of this two-years field study was to evaluate the effects of fresh and field-aged biochar on As, Cd, and Pb bioaccumulation, and on As speciation, in rice grain produced in different growing environments (flooding versus sprinkler and conventional tillage versus direct seeding). Biochar produced from holm-oak pruning residues (pyrolysis at 550 °C, 48 h), in a single application (28 Mg ha-1), reduced As bioaccumulation in rice grain in the permanent flooding system to non-quantifiable concentrations (e.g., from 0.178 mg kg-1 to <0.04 mg kg-1, for inorganic-As, respectively), an effect which remained under field-aging conditions, increasing rice commercial value. When adopting sprinkler irrigation, the undesirable increase in Cd bioaccumulation in rice, relatively to the anaerobic system, was counteracted by biochar application, reducing its bioaccumulation in kernels between 32 and 80 %, allowing a simultaneous control of metals and metalloids bioaccumulation in rice. The bioaccumulation of Pb was also prevented with biochar application, with a reduction in its concentration four- to 13-times, in all the management systems, relatively to the non-amended plots, under fresh biochar effects. However, Pb immobilization decreased with biochar field-aging, indicating that the biochar application may have to be repeated to maintain the same beneficial effect. Therefore, the present study shows that the implementation of sprinkler irrigation with holm-oak biochar could reduce the risk of heavy metals(loids) bioaccumulation in rice grains and, thereby, ensuring food safety aspects, particularly under fresh biochar effects.

Single-cell ICP-MS for studying the association of inorganic nanoparticles with cell lines derived from aquaculture species.

The current research deals with the use of single-cell inductively coupled plasma-mass spectrometry (scICP-MS) for the assessment of titanium dioxide nanoparticle (TiO2 NP) and silver nanoparticle (Ag NP) associations in cell lines derived from aquaculture species (sea bass, sea bream, and clams). The optimization studies have considered the avoidance of high dissolved background, multi-cell peak coincidence, and possible spectral interferences. Optimum operating conditions were found when using a dwell time of 50 µs for silver and 100 µs for titanium. The assessment of associated TiO2 NPs by scICP-MS required the use of ammonia as a reaction gas (flow rate at 0.75 mL min-1) for interference-free titanium determinations (measurements at an m/z ratio of 131 from the 48Ti(NH)(NH3)4 adduct). The influence of other parameters such as the number of washing cycles and the cell concentration on accurate determinations by scICP-MS was also fully investigated. Cell exposure trials were performed using PVP-Ag NPs (15 and 100 nm, nominal diameter) and citrate-TiO2 NPs (5, 25, and 45 nm, nominal diameter) at nominal concentrations of 10 and 50 µg mL-1 for citrate-TiO2 NPs and 5.0 and 50 µg mL-1 for PVP-Ag NPs. Results have shown that citrate-TiO2 NPs interact with the outer cell membranes, being quite low in the number of citrate-TiO2 NPs that enters the cells (the high degree of aggregation is the main factor which leads to the aggregates being in the extracellular medium). In contrast, PVP-Ag NPs have been found to enter the cells.

Medium-Term Effects of Sprinkler Irrigation Combined with a Single Compost Application on Water and Rice Productivity and Food Safety.

Traditional rice (Oryza sativa L.) management (tillage and flooding) is unsustainable due to soil degradation and the large amount of irrigation water used, an issue which is exacerbated in the Mediterranean region. Therefore, there is a need to explore rice management strategies in order to improve water-use efficiency and ensure its sustainability. Thus, field experiments were conducted to determine the medium-term effects of different irrigation and tillage methods combined with a single compost application on water and rice productivity, as well as food safety in a semiarid Mediterranean region. The management systems evaluated were: sprinkler irrigation in combination with no-tillage (SNT), sprinkler irrigation in combination with conventional tillage (ST), which were implemented in 2015, and flooding irrigation in combination with conventional tillage (FT), and their homologues (SNT-C, ST-C, and FT-C) with single compost application in 2015. In reference to rice grain yield, the highest values were observed under ST treatment with 10 307 and 11 625 kg ha-1 in 2018 and 2019 respectively; whereas between FT and SNT there were no significant differences, with 8 140 kg ha-1 as mean value through the study. Nevertheless, sprinkler irrigation allowed saving 55% of the total amount of water applied in reference to flooding irrigation. Furthermore, the highest arsenic concentration in grains was found under FT but it decreased with compost application (FT-C) and especially with sprinkler irrigation, regardless of tillage management systems. However, sprinkler irrigation favors the cadmium uptake by plants, although this process was reduced under SNT in reference to ST, and especially under amended compost treatments. Therefore, our results suggested that a combination of sprinkler irrigation and compost application, regardless of the tillage system, could be an excellent strategy for rice management for the Mediterranean environment in terms of water and crop productivity as well as food safety.

Combined use of biochar and alternative management systems for imazamox induced pollution control in rice growing environments.

Imazamox (IMZX) is a persistent herbicide having probable risks for non-target organisms in the environment and water contamination. Alternatives to conventional flooding rice production, including biochar amendment, may induce changes in soil properties which can greatly modify the environmental fate of IMZX. This two-year study is the first to evaluate how tillage and irrigation practices, with or without fresh or aged biochar (Bc), that are alternatives to conventional rice production impact IMZX's environmental fate. The treatments were: conventional tillage and flooding irrigation (CTFI), conventional tillage and sprinkler irrigation (CTSI), no-tillage and sprinkler irrigation (NTSI), and the corresponding Bc-amendment treatments (CTFI-Bc, CTSI-Bc, and NTSI-Bc). Fresh and aged Bc amendment decreased IMZX's sorption onto the soil in tillage treatments, with Kf values decreasing 3.7 and 4.2-fold (fresh case) and 1.5 and 2.6-fold (aged case) for CTSI-Bc and CTFI-Bc, respectively. The transition to sprinkler irrigation reduced IMZX persistence. Overall, Bc amendment also reduced chemical persistence with half-life values decreasing 1.6 and 1.5-fold for CTFI and CTSI (fresh year) and 1.1, 1.1, and 1.3-fold for CTFI, CTSI, and NTSI (aged year), respectively. Sprinkler irrigation reduced IMZX leaching by up to a factor of 2.2. The use of Bc as amendment led to a significant decrease in IMZX leaching only under tillage conditions, but notable in particular for the CTFI case where leaching losses were reduced in the fresh year from 80% to 34% and, in the aged year, from 74% to 50%. Hence the change in irrigation from flooding to sprinkler either alone or in combination with the use of Bc (fresh or aged) amendment could be considered an effective way to sharply mitigate IMZX contamination of water in environments where rice is grown, particularly in those managed with tillage.

Isolation and quantification of synthetic cannabinoid receptor agonists in human urine using membrane-assisted solvent extraction followed by liquid chromatography-tandem mass spectrometry.

The global market for new psychoactive substances (NPSs) continues to expand, and the range of drugs available on the market has probably never been wider. Synthetic cannabinoids (SCRAs) constitute the largest family of NPSs, and they go unnoticed during illicit drug market control and during routine toxicological-forensic analysis. Membrane-assisted solvent extraction (MASE) has been a novelty proposed for the simultaneous extraction of SCRAs, and urine has been selected as a model forensic-clinical sample. Isolated SCRAs were further determined by liquid chromatography-tandem mass spectrometry (LC-MS/MS). An optimised sample pre-treatment procedure consists of using 400 µL of n-hexane as an extraction phase placed inside a polypropylene (PP) membrane, adjusting the donor phase (urine) at a pH value of 5.9. Extraction was assisted by mechanical (orbital-horizontal) stirring in a temperature-controlled chamber at room temperature for 20 min. n-Hexane extracts were evaporated to dryness and re-suspended in 100 µL of mobile phase, which leads to a pre-concentration factor of 50. Method validation showed analytical recoveries higher than 80% for most SCRAs and repeatability (inter-day and intra-day assays) with RSD values lower than 20%. The proposed method was found to be selective and sensitive and limits of quantification (LOQs) between 0.10 and 1.0 µg L-1 were achieved.

PARK7/DJ-1 inhibition decreases invasion and proliferation of uveal melanoma cells.

INTRODUCTION: PARK7/DJ-1 is an oncogene that is associated with tumorigenesis in many cancers. Recent studies have demonstrated the importance of DJ-1 in the origin and development of uveal melanoma (UM). We present an analysis of the role of the DJ-1 protein in UM cells, especially in its effect on proliferation and migration. METHODS: UM cells from a primary tumor, Mel 270, and its liver metastasis, OMM2.5, were transfected with lentiviral-delivered shRNA against PARK7/DJ-1. Evaluation of cell migration and proliferation was performed using the xCELLigence real-time cell analyzer (RTCA). The effect of DJ-1 inhibition on the PTEN-Akt signaling pathway was also studied by immunoblotting. RESULTS: The silencing of PARK7/DJ-1 oncoprotein expression produced a significant decrease of phosphorylated Akt (S473) in Mel270 and in metastatic OMM2.5 UM cells with no alteration on tumor suppressor PTEN expression. The diminution of PARK7/DJ-1 expression significantly inhibited real-time proliferation and invasion of Mel270 and OMM2.5 and the invasion potential of the metastatic cells. CONCLUSION: DJ-1 appears to play a key role on the PTEN/Akt pathway in UM. DJ-1 inhibition appears to have a negative effect on proliferation and invasion of UM cells. This suggests DJ-1 as a potential therapeutic target in UM.

Effects of Combined Use of Olive Mill Waste Compost and Sprinkler Irrigation on GHG Emissions and Net Ecosystem Carbon Budget under Different Tillage Systems.

Traditional rice (Oryza sativa L.) production by flooding is a source of greenhouse gases (GHG), especially methane. The high consumption of water, as well as the chemical and physical degradation caused by these traditional practices in rice soils, is promoting a decrease in rice production in the Mediterranean area. The aim of this study was to monitor GHG emissions and the net ecosystem carbon balance (NECB) from rice produced with sprinkler irrigation techniques and also assess the impact of olive mill waste compost (C-OW) application and tillage on GHG emissions and the NECB. A field experiment for irrigated rice production was implemented by considering four different treatments: (1) tillage (T); (2) no tillage-direct seeding techniques (DS); (3) application of C-OW followed by tillage (TC); and (4) application of C-OW followed by direct seeding (DSC). The C-OW was only applied in the first year at a dose of 80 Mg ha-1. GHG emissions were monitored over three years in these four treatments in order to estimate the direct (first year) and residual (third year) effects of such practices. The application of C-OW caused an increase of 1.85 times the emission of CO2-C in the TC-DSC compared to the T-DS in the first year. It is noteworthy that the TC treatment was the only one that maintained an emission of CO2-C that was 42% higher than T in the third year. Regardless of the treatments and year of the study, negative values for the cumulative CH4 were found, suggesting that under sprinkler irrigation, CH4 oxidation was the dominant process. A decrease in N2O emissions was observed under direct seeding relative to the tillage treatments, although without significant differences. Tillage resulted in an increase in the global warming potential (GWP) of up to 31% with respect to direct seeding management in the third year, as a consequence of the greater carbon oxidation caused by intensive tillage. DS presented a positive NECB in the accumulation of C in the soil; therefore, it provided a greater ecological benefit to the environment. Thus, under Mediterranean conditions, rice production through a sprinkler irrigation system in combination with direct seeding techniques may be a sustainable alternative for rice crops, reducing their GWP and resulting in a lower carbon footprint. However, the use of C-OW as an organic amendment could increase the GHG emissions from rice fields irrigated by sprinklers, especially under tillage conditions.

Environmental fate and efficiency of bispyribac­sodium in rice soils under conventional and alternative production systems affected by fresh and aged biochar amendment.

Irrigation and tillage practice alternatives to conventional flooding production, with or without organic amendments, are attracting great interest to adapt rice cultivation to climate change. However, they can alter the behaviour of pesticides and their efficiency against weeds. A two-year field experiment was conducted to investigate how the environmental fate and the weed control efficiency (WCE) of bispyribac­sodium (BS) were influenced by biochar produced from holm oak prunings (BHO) testing both the fresh and the aged effects. The treatments were: flooding irrigation and tillage (FT), sprinkler irrigation and tillage (ST), sprinkler irrigation and no-tillage (SNT), and the corresponding homologues with BHO addition (FT-BHO, ST-BHO, and SNT-BHO, respectively). Fresh BHO amendment decreased the sorption of BS onto the soil in all treatments, while, after aging, it also decreased sorption in FT-BHO (1.3-fold) but increased it in SNT-BHO and ST-BHO (1.1-fold). BHO addition reduced BS persistence under non-flooding and flooding incubation conditions, except for FT under the former condition for which t1/2 increased ≈1.5-fold in both years. The addition of BHO led to a decrease in BS leaching from 58.3 % and 44.6 % and from 70.4 % and 58.1 % in ST and FT to 50.1 % and 38.3 % and 63.6 % and 50.3 % in the homologue amended soils for the fresh and aged years, respectively. While fresh BHO addition decreased the WCE of BS in SNT-BHO, ST-BHO, and FT-BHO on average by a factor of 1.5, with aged BHO there was only such a decrease (by a factor of 1.4) in FT-BHO. The use of BHO could be effective for reducing water contamination by BS in flooding or sprinkler irrigation rice farming as long as conventional tillage is used. But it may also contribute to greatly reducing the herbicide's efficiency, although with time to allow aging, this reduction would only persist under conventional flooding production.

Ultrasonication followed by enzymatic hydrolysis as a sample pre-treatment for the determination of Ag nanoparticles in edible seaweed by SP-ICP-MS.

Seaweed can bioaccumulate nanomaterials that would be transferred to the trophic chain. This work describes the optimization of a method for the separation of silver nanoparticles (AgNPs) from seaweed using an ultrasound-assisted enzymatic hydrolysis method and ulterior determination by single particle inductively coupled plasma mass spectrometry (SP-ICP-MS). The following parameters affecting the isolation of AgNPs were optimized using a Palmaria palmata (red seaweed) sample previously exposed to AgNPs: type of sonication (bath vs. ultrasonic probe), ultrasound amplitude, sonication time, sonication mode (pulsed vs. continuous sonication), concentration of the enzymes mixture (Macerozyme R-10®), and enzymatic hydrolysis time. The stability of AgNPs during extraction was tested by transmission electron microscopy (TEM) and using a standard of 15 nm of polyvinylpyrrolidone (PVP)-coated AgNPs analyzed by SP-ICP-MS. The analytical performance was evaluated with good results. For total Ag determination, the limits of detection and quantification were 2.2 and 7.7 ng g-1, respectively; and for AgNPs determination, the limits of detection in size and number were 14 nm and 4.34 × 107 part g-1, respectively. Besides, the matrix effect, the repeatability and the analytical recovery were also studied. Finally, the method was applied to the analysis of several red (Palmaria palmata) and green (Ulva sp.) seaweed samples.

Applications of Non-Coding RNAs in Patients With Retinoblastoma.

Retinoblastoma (RB) is the most common primary intraocular malignancy in childhood. In the carcinogenic process of neoplasms such as RB, the role of non-coding RNAs (ncRNAs) has been widely demonstrated recently. In this review, we aim to provide a clinical overview of the current knowledge regarding ncRNAs in relation to RB. Although ncRNAs are now considered as potential diagnostic biomarkers, prognostic factors, and therapeutic targets, further studies will facilitate enhanced understanding of ncRNAs in RB physiopathology and define the roles ncRNAs can play in clinical practice.

Combined use of olive mill waste compost and sprinkler irrigation to decrease the risk of As and Cd accumulation in rice grain.

Sprinkler irrigation has been successfully introduced in rice production as an alternative to the traditional flooding system, allowing water savings and the reduction of As accumulation in the grain. However, the same conditions can increase Cd mobility and grain accumulation, an effect that needs to be countered. A 3-year field experiment was set-up in a Mediterranean region (Extremadura, Spain), to evaluate how the application of compost from olive mill waste (single application, 80 t ha-1), influences the accumulation of As and Cd in the grain under different irrigation regimes. Accumulation of As in the grain was always lower in the sprinkler irrigation when compared with the flooding irrigation, reaching a 5-fold difference in the third year. Compost application did not evidence a clear effect on the As accumulation in the rice grain, but highly significant negative correlations (p < 0.001) were obtained between As content in the grain (total, inorganic, and organic) and the humification parameters in the soil, evidencing the importance of using a mature and stable organic amendment to avoid As accumulation in the grain. Cadmium accumulation in the rice grain decreased in each treatment where compost was applied, relatively to the non-treated counterpart (e.g., from 0.080 to <0.010 mg kg-1, in direct seeding with sprinkler irrigation, in the third year). There were no significant differences in the total inorganic As between treatments with or without compost application, but it was possible to observe an increase in the predominance of the organic As over the more toxic inorganic As, when compost was applied, allowing a decrease in the risk associated to As accumulation. Therefore, the aerobic cultivation of rice, with the simultaneous application of an adequate source of organic matter, can be considered a good solution to cope with the risk of accumulation of As and Cd in the rice grain.

Antibodies against 4 Atypical Post-Translational Protein Modifications in Patients with Rheumatoid Arthritis.

Patients with rheumatoid arthritis (RA) show autoantibodies against post-translational protein modifications (PTMs), such as anti-citrullinated protein antibodies. However, the range of recognized PTMs is unknown. Here, we addressed four PTMs: chlorination, non-enzymatic glycation, nitration, and homocysteinylation, identified as targets of atypical RA autoantibodies in studies whose protocols we have followed. The modified antigens included collagen type II, an extract of synovial proteins and a selection of peptides. We interpreted the results according to the optical density (OD) obtained in an enzyme-linked immunosorbent assay ( ELISA) with the modified antigen and the corrected OD obtained after subtracting the reactivity against the unmodified antigen. The results showed evidence of specific antibodies against glycated collagen type II, as the corrected ODs were higher in the 182 patients with RA than in the 164 healthy controls (p = 0.0003). However, the relevance of these antibodies was doubtful because the magnitude of the specific signal was small (median OD = 0.072 vs. 0.027, respectively). There were no specific antibodies against any of the other three PTMs. Therefore, our results showed that the four PTMs are not inducing a significant autoantibody response in patients with RA. These results indicated that the repertoire of PTM autoantigens in RA is restricted.

Single-particle inductively coupled plasma mass spectrometry using ammonia reaction gas as a reliable and free-interference determination of metallic nanoparticles.

Intensive production of nanomaterials, especially metallic nanoparticles (MNPs), and their release into the environment pose several risks for humans and ecosystem health. Consequently, high-efficiency analytical methodologies are required for control and characterization of these emerging pollutants. Single-particle inductively coupled plasma - mass spectrometry (SP-ICP-MS) is a promising technique which allows the determination and characterization of MNPs. However, several elements or isotopes are hampered by spectral interferences, and dynamic-reaction cell (DRC) technology is becoming a useful tool for free interference determination by ICP-MS. DRC-based SP-ICP-MS methods using ammonia as a reaction gas (either on-mass approach or mass-shift approaches) have been developed for determining titanium dioxide nanoparticles (TiO2 NPs), copper oxide nanoparticles (CuO NPs), copper nanoparticles (Cu NPs), and zinc oxide nanoparticles (ZnO NPs). The effects of parameters such as ammonia flow rate and dwell time on the peak width (NP transient signal in SP-ICP-MS) were comprehensively studied. Influence of NP size and nature were also investigated.