Fibrin-Plasma Rich in Growth Factors Membrane for the Treatment of a Rabbit Alkali-Burn Lesion.

The purpose of this work is to describe the use of Fibrin-Plasma Rich in Growth Factors (PRGF) membranes for the treatment of a rabbit alkali-burn lesion. For this purpose, an alkali-burn lesion was induced in 15 rabbits. A week later, clinical events were evaluated and rabbits were divided into five treatment groups: rabbits treated with medical treatment, with a fibrin-PRGF membrane cultured with autologous or heterologous rabbit Limbal Epithelial Progenitor Cells (LEPCs), with a fibrin-PRGF membrane in a Simple Limbal Epithelial Transplantation and with a fibrin-PRGF membrane without cultured LEPCs. After 40 days of follow-up, corneas were subjected to histochemical examination and immunostaining against corneal or conjunctival markers. Seven days after alkali-burn lesion, it was observed that rabbits showed opaque cornea, new blood vessels across the limbus penetrating the cornea and epithelial defects. At the end of the follow-up period, an improvement of the clinical parameters analyzed was observed in transplanted rabbits. However, only rabbits transplanted with cultured LEPCs were positive for corneal markers. Otherwise, rabbits in the other three groups showed positive staining against conjunctival markers. In conclusion, fibrin-PRGF membrane improved the chemically induced lesions. Nonetheless, only fibrin-PRGF membranes cultured with rabbit LEPCs were able to restore the corneal surface.

Ultrastructural Alterations of Midgut Epithelium, But Not Greater Wing Fluctuating Asymmetry, in Paper Wasps (Polistes dominula) from Urban Environments.

Polistes paper wasps can be used to monitor trace metal contaminants, but the effects of pollution on the health of these insects are still unknown. We evaluated, in a south-eastern area of Spain, whether workers of Polistes dominula collected at urban and rural sites differ in health of midgut tissue and in fluctuating asymmetry, an estimate of developmental noise. We found that wasps collected at the urban sites had abundant lead (Pb)-containing spherites, which were less visible in wasps from the rural sites. Evident ultrastructural alterations in the epithelium of the midgut of the wasps collected at the urban sites included broken and disorganized microvilli, a high amount and density of heterochromatin in the nucleus of epithelial cells, cytoplasmic vacuolization and mitochondrial disruptions. Altogether, these findings suggest a negative effect on the transmembrane transport and a less efficient transcription. On the contrary, a healthy epithelium was observed in wasps from the rural sites. These differences may be preliminarily linked with levels of lead pollution, given that wasps from urban sites had double the Pb concentrations of wasps from rural sites. Level of fluctuating asymmetry was unrelated to wasp origin, thus suggesting no link between developmental noise and Pb-driven pollution.

Use of a versatile, easy, and rapid atmospheric monitor (VERAM) passive samplers for pesticide determination in continental waters.

The versatile, easy, and rapid atmospheric monitor (VERAM), a passive sampler device widely used for air monitoring, was evaluated as passive sampler for the determination of 23 pesticides in water. Gas chromatography with mass spectrometry detection was employed for determination of pesticides after microwave-assisted-extraction and specific clean-up of deployed samplers. The proposed methodology reached method detection levels from 2 to 10 ng pesticide per sampler. Sampling rate (Rs) was determined for every pesticide from an uptake isotherm study performed at three different concentration levels (50, 125, and 250 ng L-1). The obtained RS values ranged from 0.06 to 0.76 L d-1. The obtained limits of detection for a 24-h passive sampling were from 4 to 50 ng L-1. The effect of water parameters, such as temperature, pH, and ionic strength, were evaluated for their effect on pesticides retention using VERAMs. Pesticide RS values were independent of the water composition and increased on increasing temperature. Finally, the VERAM uptake was compared with that obtained using classic semipermeable membrane devices (SPMDs). This study is the first precedent for the use of VERAMs as passive samplers for the adsorption and concentration of pesticides in water and it confirms the satisfactory analytical figures of merit of VERAM as passive sampler of water. Graphical Abstract Scheme of water sampling of pesticides using versatile, easy, and rapid atmospheric monitor (VERAM) passive samplers.

Rare earth elements sediment analysis tracing anthropogenic activities in the stratigraphic sequence of Alagankulam (India).

A methodological approach based on rare earth elements analysis was developed to observe human activities in the stratigraphic sequence of Alagankulam. The site was one of the main ancient ports in south-eastern India and one of the transoceanic connecting points between East and West during the Classical Period. The sampled sediments where collected from vertical profiles, areas with traces of firing activities and filled deposits. Major, minor and trace element concentrations were measured by the means of spectroscopic and spectrometric techniques. Data from multielemental analysis were then cross-referenced together with archaeological evidence to map the variability within the site and its association with the detected anthropic activities. The matching of the interpretation of the archaeological record and the analytical data has allowed a combined mapping of visible and invisible traces of human activities in the site, giving a deeper insight of the Alagankulam occupational history.

The casts of Pompeii: Post-depositional methodological insights.

The casts of Pompeii bear witness to the people who died during the Vesuvius 79 AD eruption. However, studies on the cause of death of these victims have not been conclusive. A previous important step is the understanding of the post-depositional processes and the impact of the plaster in bones, two issues that have not been previously evaluated. Here we report on the anthropological and the first chemical data obtained from the study of six casts from Porta Nola area and one from Terme Suburbane. A non-invasive chemical analysis by portable X-ray fluorescence was employed for the first time on these casts of Pompeii to determine the elemental composition of the bones and the plaster. Elemental profiles were determined providing important data that cross-referenced with anthropological and stratigraphic results, are clearly helpful in the reconstruction of the perimortem and post-mortem events concerning the history of these individuals. The comparative analyses carried out on the bone casts and other collections from burned bones of the necropolis of Porta Nola in Pompeii and Rome Sepolcreto Ostiense, and buried bones from Valencia (Spain), reveal the extent of high temperature alteration and post-depositional plaster contamination. These factors make bioarchaeological analyses difficult but still allow us to support asphyxia as the likely cause of death.

Biomonitoring of polycyclic aromatic hydrocarbons in the urine of lactating mothers: Urinary levels, association with lifestyle factors, and risk assessment.

Polycyclic aromatic hydrocarbons (PAHs) are pollutants produced during incomplete combustion of organic matter and several industrial processes. Humans can be exposed to PAHs through ingestion of food, inhalation of tobacco smoke or polluted air, and dermal contact, causing immunologic, developmental, and reproductive problems. In the present research, eleven metabolites of PAHs were analyzed in the urine of 110 lactating women living in Spain (2015). PAH metabolites were extracted from the urine samples by liquid-liquid extraction and their determination was performed by liquid chromatography coupled to tandem mass spectrometry. In addition, information on lifestyle and dietary habits of the participants was collected using a questionnaire. All the PAH metabolites were detected in more than 70% of the samples, except for 3-hydroxybenzo(a)pyrene which was detected in less than 1% of the samples. The highest urinary levels were found for naphthalene metabolites, with geometric means of 0.8 (1-hydroxynaphthalene) and 7.1 ng ml-1 (2-hydroxynaphthalene). The statistical analysis showed that smoking status, as well as the ingestion of certain food groups (vegetables, cereals, oils and fats, smoked fish and coffee), were the main influencing factors of exposure to PAHs. The estimated daily intake (EDI) was calculated for naphthalene, fluorene, phenanthrene, and pyrene, ranging from 6 to 1522 ng kg-1·day-1. The non-cancer risk associated to PAH exposure was estimated, showing hazard quotients (HQs) and hazard indexes (HIs) below 1. Therefore, it did not reveal a significant health risk for Spanish women due to PAH exposure.

Determination of Third-Generation Synthetic Cannabinoids in Oral Fluids.

A procedure has been developed for the determination of third-generation synthetic cannabinoids in oral fluid samples by using a semi-automated microextraction by packed sorbent (MEPS) procedure and gas chromatography-mass spectrometry (GC-MS) determination. Five synthetic cannabinoids were employed as model compounds 5F-ADB, MMB-CHMICA, THJ-2201, CUMYL-4CN-BINACA and MDMB-CHMCZCA. The most adequate operative conditions for MEPS were evaluated giving quantitative recoveries, from 89 to 124%, in synthetic and field saliva samples spiked with 125 and 250 µg/L of the studied cannabinoids, with the exception of MDMB-CHMCZCA in field saliva samples that provided slightly lower recoveries from 62 to 66%. A high sensitivity was obtained for the proposed MEPS-GC-MS procedure with limits of detection from 10 to 20 µg/L. The obtained results demonstrate the high potential of MEPS-GC-MS combination for semi-automated, selective and sensitive determination of synthetic cannabinoids in oral fluid samples.

Electroanatomic study of the left atrial insertion of an epicardial accessory pathway integrating the coronary sinus.

Endocardial electro-anatomic reconstruction of the left atrium and activation mapping defined a very large atrial accessory pathway insertion with a previously failed ablation attempt. Radiofrequency application inside the coronary sinus (CS), at a site with a sharp electrogram bridging atrial and ventricular electrograms abolished pathway conduction. The myocardium in the venous branches of the CS appeared to be responsible for this extraordinary atrial insertion area.

A simple pacing method to diagnose postero-anterior (clockwise) cavo-tricuspid isthmus block after radiofrequency ablation.

AIMS: Bidirectional block of the cavo-tricuspid isthmus (CTI) is a widely accepted endpoint for typical atrial flutter ablation, but its evaluation may be difficult, especially in the postero-anterior (clockwise) direction. The main goal was to evaluate pacing at the septal edge of the ablation line as an indicator of clockwise CTI block and as a predictor for flutter recurrence. METHODS AND RESULTS: In 94 patients undergoing flutter ablation, CTI block in the antero-posterior (counterclockwise) direction was determined by differential pacing from several levels of the anterior right atrial (RA). CTI block in the clockwise direction was evaluated by analysing electrograms (EGM) at the ablation line during differential pacing of the septal RA (differential septal pacing) or by anterior sequence of RA during pacing septal isthmus, next to the ablation line (septal CTI pacing). Ablation produced bidirectional block in 78% of the patients, unidirectional counterclockwise block in 9% and bidirectional conduction persisted in 13%. After follow-up (37 +/- 23 months), flutter recurrence occurred in 13% (48% if persistent conduction vs. 3% if bidirectional block, P < 0.001). During differential septal pacing, EGMs were difficult to interpret in 36% of the patients; in these cases, the diagnosis of CTI block or conduction in the clockwise direction was clearly established by using septal CTI pacing. CONCLUSION: Activation sequence of anterior RA during septal CTI pacing, next to the ablation line, is a reliable and simple method to diagnose clockwise CTI block and is associated with a low flutter recurrence.

Risk assessment of exposure to phthalates in breastfeeding women using human biomonitoring.

In this study, we assessed the presence of 14 phthalate metabolites in the urine of 104 lactating mothers from Valencia (Spain) who took part in the human biomonitoring project BETTERMILK. Nine of the metabolites studied showed detection frequencies >80%, whereas the rest of the metabolites presented low detection frequencies (<5%). The concentrations ranged from

Biomonitoring of parabens in human milk and estimated daily intake for breastfed infants.

In this study, we assessed the presence of four parabens in human milk of 120 mothers from Valencia (Spain) which took part in a human biomonitoring project (BETTERMILK). The detection frequency ranges of parabens were 41-60% and 61-89% for unconjugated- and total (unconjugated + conjugated)-parabens, respectively. The concentrations ranged from

Biomonitoring of non-persistent pesticides in urine from lactating mothers: Exposure and risk assessment.

The aim of the present study was to assess the exposure to pesticides in urine from Spanish lactating mothers (n = 116). Six nonspecific (dialkyl phosphates) and 20 specific metabolites of organophosphate pesticides (OPs), herbicides and pyrethroids were analyzed. The most frequently detected biomarkers were diethyl phosphate, p-nitrophenol, 3,5,6-trichloro-2-pyridinol and 3-phenoxybenzoic acid, whose geometric means were 1.9 ng·mL-1, 0.8 ng·mL-1, 1.5 ng·mL-1 and 1.4 ng·mL-1, respectively. Herbicide metabolites were the least frequently detected biomarkers with detection frequencies between 0% (2,4,5-Trichlorophenoxyacetic acid) and 22% (2,4-Dichlorophenoxyacetic acid). Multiple regression analyses showed that the closeness to a farming activity, the place of residence and the presence of garden/plants at home were some of the most important contributors to urinary levels of pesticide metabolites. Estimated daily intake (EDI), hazard quotient (HQ) and hazard index (HI) were obtained in order to interpret urinary levels of the most frequently detected pesticide metabolites in a risk assessment context. The highest EDIs were obtained for chlorpyrifos (0.40-1.14 µg·kg bw-1·day-1) and deltamethrin (0.34-4.73 µg·kg bw-1·day-1). The calculated HQ for chlorpyrifos, dimethoate, parathion and deltamethrin ranged from 0.01 to 0.47, and HI for OPs ranged from 0.09 to 0.33 showing that apparently there were low health risks due to the exposure to these pesticides in this group of Spanish breastfeeding women.

Use of semipermeable membrane devices for monitoring pesticides in indoor air.

In this work, 40 pesticides from different categories were analyzed in the air of 20 indoor places that have an intensive use of pesticides. Passive sampling was carried out by using semipermeable membrane devices (SPMDs) deployed for 7 days. SPMDs were analyzed using microwave-assisted extraction and GC/MS. PCB-195 was used as an internal standard. Recoveries ranged between 81 and 108% for many compounds, with RSD values < 11%. Typical LOD values for 7 days of sampling were from 0.1 to 3.1 ng/m. Propamocarb, propoxur, carbosulfan, pirimicarb, metribuzin, metalaxyl, pendimethalin, oxadiazon, phenothrin, and permethrin were detected in 11 sampling sites, with air concentrations in the range from 1 to 921 ng/m. However, the pesticide levels found cannot be considered a serious health problem according to results from other pesticide monitoring studies.

Optimization of Resolving Power, Fragmentation, and Mass Calibration in an Orbitrap Spectrometer for Analysis of 24 Pesticide Metabolites in Urine.

Mass spectrometer parameters such as Resolving Power, type of fragmentation, and mass calibration mode were optimized in the analysis of 24 pesticide metabolites in human urine using Ultra-High Pressure Liquid Chromatography coupled to Orbitrap High-Resolution Mass Spectrometer (UHPLC-HRMS). The best results were achieved with a Resolving Power of 25,000 FWHM and by applying Collision Induced Dissociation fragmentation mode (40 eV).

Development of a novel methodology for determination of dialkyl phosphates in human urine using liquid chromatography-tandem mass spectrometry.

In this study a fast and sensitive method was developed for the quantitative determination of six dialkyl phosphates (DAPs) in human urine using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The proposed methodology was based on liquid-liquid extraction (LLE) with diethyl ether and ethyl acetate using MgSO4 and NaCl, followed by liquid chromatography and mass spectrometry detection in the selected reaction mode (SRM) with negative electrospray ionization (ESI-). Detection settings were optimized by design of experiments (DoE). Dibutyl phosphate (DBP) was used as internal standard. Several criteria established in the SANTE/11813/2017 guidance document for pesticide residues and analysis in food and feed were used to validate the suggested method: recoveries (R %) between 70 and 120% and coefficients of variation (CV %) ≤ 20%. The procedural limit of quantification (LoQ) was 0.50 ng mL-1 for the six DAPs. The R% and CV% ranges were 82-117% and 3-20%, respectively. In total, 20 spot urine samples from lactating mothers were satisfactorily analyzed by the proposed analytical strategy.

Determination of four parabens and bisphenols A, F and S in human breast milk using QuEChERS and liquid chromatography coupled to mass spectrometry.

For the first time, a multiresidue, sensitive and high throughput method for determination of bisphenol A, F and S and 4 parabens (methyl paraben, ethyl paraben, propyl paraben and butyl paraben) in human breast milk was developed. The proposed method includes an extraction and clean-up procedure based on QuEChERS methodology followed by liquid chromatography coupled to triple quadrupole mass spectrometry determination. Negative atmospheric pressure chemical ionization in the selected reaction monitoring mode was used for mass detection. During the method validation the recoveries varied between 83 and 115% with a precision lower than 20% for all analytes using spiked levels from 0.1 to 50 ng mL-1. The LOQ was 0.10 ng mL-1 for most of the analytes. The proposed method was successfully applied for the determination of these compounds in 10 breast milk samples from volunteer lactating mothers from the Valencian region (Spain). Among parabens, methyl paraben presented the highest detection frequency (80%) with a concentration range of 0.11-7.00 ng mL-1, while bisphenol A was detected more frequently than BPF and BPS (80% of detection frequency) with concentrations ranging from 0.13 to 1.62 ng mL-1.

Biomonitoring of bisphenols A, F, S in human milk and probabilistic risk assessment for breastfed infants.

The present study addresses the presence of bisphenols A (BPA) and its analogs bisphenol F (BPF) and S (BPS) in milk of 120 mothers living in Valencia (Spain) and participating in the BETTERMILK project (year 2015). We also studied the factors that could influence the BPA levels and estimated the exposure and the risk for breast fed infants. The frequency of detection of total (conjugated + unconjugated) and unconjugated-BPA were 83% and 77%, with a geometric mean of 0.29 ng/mL and 0.15 ng/mL, respectively. The frequency of detection was much lower for total-BPF (22%) and total-BPS (1.1%). The place of residence of the mother and the use of personal care products showed significant association with BPA concentrations. The estimated daily intake of total-BPA for breastfed infants amounted to a geometric mean of 0.04 µg/kg bw and a 95th percentile of 1.0 µg/kg bw, below the tolerable daily intake of 4 µg/kg bw-day established by EFSA. To our knowledge, this is the largest biomonitoring study of bisphenols in human milk in Europe.

Spanish Results of the Second European Cardiac Resynchronization Therapy Survey (CRT-Survey II).

INTRODUCTION AND OBJECTIVES: We describe the results for Spain of the Second European Cardiac Resynchronization Therapy Survey (CRT-Survey II) and compare them with those of the other participating countries. METHODS: We included patients undergoing CRT device implantation between October 2015 and December 2016 in 36 participating Spanish centers. We registered the patients' baseline characteristics, implant procedure data, and short-term follow-up information until hospital discharge. RESULTS: Implant success was achieved in 95.9%. The median [interquartile range] annual implantation rate by center was significantly lower in Spain than in the other participating countries: 30 implants/y [21-50] vs 55 implants/y [33-100]; P=.00003. In Spanish centers, there was a lower proportion of patients ≥ 75 years (27.9% vs 32.4%; P=.0071), a higher proportion in New York Heart Association functional class II (46.9% vs 36.9%; P <.00001), and a higher percentage with electrocardiographic criteria of left bundle branch block (82.9% vs 74.6%; P <.00001). The mean length of hospital stay was significantly lower in Spanish centers (5.8±8.5 days vs 6.4±11.6; P <.00001). Spanish patients were more likely to receive a quadripolar LV lead (74% vs 56%; P <.00001) and to be followed up by remote monitoring (55.8% vs 27.7%; P <.00001). CONCLUSIONS: The CRT-Survey II shows that, compared with other participating countries, fewer patients in Spain aged ≥ 75 years received a CRT device, while more patients were in New York Heart Association functional class II and had left bundle branch block. In addition, the length of hospital stay was shorter, and there was greater use of quadripolar LV leads and remote CRT monitoring.

Analysis of currently used pesticides in fine airborne particulate matter (PM 2.5) by pressurized liquid extraction and liquid chromatography-tandem mass spectrometry.

During and after the application of currently used pesticides (CUPs) a significant fraction of applied pesticides can be lost to the air. A confirmatory and rapid procedure has been developed for the determination of four fungicides (carbendazim, thiabendazol, imazalil and bitertanol), three insecticides (imidacloprid, methidathion and pyriproxyfen), one helicide (methiocarb) and one acaricide (hexythiazox) in fine airborne particulate matter (PM 2.5) at trace level. The proposed method includes extraction of PM 2.5-bound pesticides by pressurized liquid extraction (PLE) followed by a direct injection into LC-MS/MS. The main parameters affecting the performance of the electrospray ionization source and PLE parameters were optimised using statistical design of experiments (DoE). The matrix effect was also evaluated. Recoveries ranged from 86 to 106% and the limit of quantification (LoQ) was 6.5 pg m(-3) for eight out of nine pesticides, when air volumes of 760 m(3) were collected. The method was applied to 60 samples collected from four stations of the monitoring network of the Regional Valencia Government (Spain) during August-October 2007. The measured concentrations ranged from not detected to 1,371 pg m(-3).