Search

Pteridines are a group of compounds synthesised by many living organisms that are involved in the metabolism of many cofactors and vitamins. Their concentration in biological fluids may be altered by various pathologies such as cancer or inflammatory bowel disease, urine being the main route of excretion. In this study, three lumazines and ten pterins were analysed in their native oxidation state using high-performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry. Their high concentration in urine samples and their good ionisation behaviour allow the use of the dilute-and-shoot method by simple filtration of the urine prior to chromatographic analysis. The final method offers excellent linearity, sensitivity and precision parameters, and a total of 135 urine samples were analysed from patients with some relevant information such as faecal calprotectin (FCP) levels, common diseases such as diabetes, hypertension and dyslipidaemia and immunological diseases such as inflammatory bowel disease (IBD). The pteridine profile was related to FCP levels without showing any correlation. In addition, pteridine levels were compared between healthy subjects and IBD, diabetic, hypertensive and dyslipidaemic patients, and significant differences were found between the two groups for some of the pteridines.

The present work is focused on the development of an analytical platform to elucidate the metabolic pathway of PTSO from onion, an organosulfur compound well-known for its functional and technological properties and its potential application in animal and human nutrition. This analytical platform consisted of the use of gas chromatography–mass spectrometry (GC-MS) and ultra-high performance liquid chromatography quadrupole with time-of-flight MS (UHPLC-Q-TOF-MS) in order to monitor volatile and non-volatile compounds derived from the PTSO. For the extraction of the compounds of interest, two different sample treatments were developed: liquid–liquid extraction (LLE) and salting-out assisted liquid–liquid extraction (SALLE) for GC–MS and UHPLC-Q-TOF-MS analysis, respectively. Once the analytical platform was optimised and validated, an in vivo study was planned to elucidate PTSO metabolisation, revealing the presence of dipropyl disulfide (DPDS) in liver samples with concentrations between 0.11 and 0.61 µg g−1. The DPDS maximum concentration in the liver was observed at 0.5 h after the intake. DPDS was also present in all plasma samples with concentrations between 2.1 and 2.4 µg mL−1. In regard to PTSO, it was only found in plasma at times above 5 h (0.18 µg mL−1). Both PTSO and DPDS were excreted via urine 24 h after ingestion., Spanish MICINN (PGC2018-098363-B-100), the University of Murcia (R-1328/2018), European Commission (FEDER/ERDF), CIBEREHD is funded by the Instituto de Salud Carlos III

OBJECTIVE: Venous hypertension associated with a primitive basal vein of Rosenthal (BVR) has been noted as the most likely cause of idiopathic subarachnoid hemor-rhage (iSAH). Other types of venous drainage variations have been scarcely studied but may further explain the cases not associated with a BVR anomaly. Our aim was to investigate if dural venous sinus (DVS) anomalies are related with iSAH. - METHODS: A total of 76 patients diagnosed with iSAH were identified from a prospectively maintained database and their angiographic findings compared with 76 patients diagnosed with aneurysmal subarachnoid hemorrhage. -RESULTS: On top of the BVR variations, our data showed a higher prevalence of transverse sinus hypoplasia (47.4% vs. 28.9%; P = 0.019), superior petrosal sinus hypoplasia (32.9% vs. 13.2%; P = 0.003), and clival plexus hyperplasia (65.8% vs. 43.4%; P = 0.005) in patients with iSAH. Analyzing by total number of angiograms, the iSAH group showed also a higher prevalence of inferior petrosal sinus hyperplasia (36.2% vs. 25%; P = 0.003). Of the patients with iSAH without a primitive BVR, 84% harbored double dagger 1 peri-mesencephalic DVS variation and the overall number of venous drainage variations was significantly higher in patients with iSAH. CONCLUSIONS: In addition to the well-documented BVR anomalies, there seems to be a significant relationship of other DVS variations in patients with iSAH. Transverse si-n us hypoplasia, superior petrosal sinus hypoplasia, inferior petrosal sinus hyperplasia, and clival plexus hyperplasia were significantly more frequent in patients with iSAH. The presence of double dagger 3 of those variations would increase the suspicion of a nonaneurysmatic subarachnoid hemorrhage and could help avoid a second angiogram.

Este artigo, de base teórico-reflexiva e originado a partir de um recorte de uma pesquisa de mestrado, promove reflexões sobre as experiências (ciber)formativas, on-line e off-line, advindas dos movimentos de trocas e diálogos docentes que se concretizaram em grupos de discussões (GD) desenvolvidos com professores(as) dos anos finais do Ensino Fundamental e que atuam no município de Valente, Estado da Bahia. Com o objetivo voltado para a construção coletiva de conhecimento no que tange ao uso das Tecnologias da Informação e Comunicação (TIC) para mediar o ensino e a aprendizagem em sala de aula e levando em conta os discursos produzidos coletivamente, problematiza-se saberes docentes, inquietações e relatos de sucesso sobre o uso pedagógicos dos artefatos tecnológicos, ao tempo em que aciona perspectivas teóricas sobre Cibercultura, práxis, racionalidade comunicativa, practicum reflexivo, no âmbito da formação do professor. O itinerário metodológico concebido ancora-se nos fundamentos da pesquisa qualitativa e foi subdividido em dois momentos: o on-line, destinado aos encontros coletivos e presenciais sob os fundamentos do GD; e o off-line, o qual correspondeu ao acontecer de minicursos em espaço (ciber)formativo. Fruto da práxis reflexiva, do estudo resultam ponderações críticas dos professores para com o uso das TIC em seus respectivos espaços e fazeres pedagógicos, admitindo ser possível redimensionar o processo de ensino e de aprendizagem com o uso dessas tecnologias. Corroborou-se, ainda, com o entendimento de ser uma formação dialógico-reflexiva de docentes, quando estruturada e experenciada de maneira coletiva, propulsora de aprendizagens colaborativas e estimuladoras da reconfiguração da atuação pedagógica.

Introducción. Las vasculitis son enfermedades graves que suelen tener un debut muy inespecífico y un diagnóstico tardío, lo que implica una alta tasa de fracaso terapéutico. Se presenta un caso de vasculitis que puede contribuir a considerar esta enfermedad como parte del diagnóstico diferencial desde las primeras etapas del proceso de atención para garantizar un abordaje integral y un inicio temprano del tratamiento, lo que reduciría la morbimortalidad asociada a estas entidades y mejoraría la tasa de éxito de los tratamientos. Presentación del caso. Mujer caucásica de 77 años que como único antecedente presentaba hipertensión arterial de dos años de evolución y quien consultó al servicio de urgencias de una institución de segundo nivel por cuadro clínico de un mes de evolución consistente en astenia, febrícula, hiporexia y pérdida de peso. La paciente fue hospitalizada para completar la historia clínica y dados los hallazgos se sospechó de una posible translocación bacteriana secundaria a una neoplasia intestinal y se inició cobertura antibiótica empírica, pero su condición seguía empeorando. Se realizaron pruebas complementarias que no arrojaron resultados concluyentes; ante la persistencia de la fiebre, la insuficiencia renal y la anemia, se realizó una biopsia de riñón que mostró vaso arterial con necrosis fibrinoide e infiltrados de polimorfonucleares asociados, claros signos de una vasculitis activa del tipo poliangeítis microscópica. Se instauró el tratamiento indicado pero la paciente tuvo una evolución desfavorable y falleció. Conclusiones. La exposición de este caso inusual contribuye a que los profesionales de la salud consideren el diagnóstico de vasculitis desde el inicio de la atención, aun cuando la sintomatología sugiere otras patologías o incluso cuando se presentan síntomas tan inespecíficos como anemia y pérdida de peso, ya que esto ayudará a establecer un diagnóstico temprano.

Background Containment policies and other restrictions introduced by the Spanish government in response to the COVID-19 pandemic present challenges for marginalised populations, such as people who use drugs. Harm reduction centres are often linked to social services, mental health services, and infectious disease testing, in addition to tools and services that help to reduce the harms associated with injecting drugs. This study aimed to explore the impact of the pandemic on these services in four autonomous communities in Spain. Methods This is a cross-sectional study that employed a seven-section structured survey administered electronically to 20 centres in July 2020. Data from the most heavily affected months (March–June) in 2020 were compared to data from the same period in 2019. Averages were calculated with their ranges, rates, and absolute numbers. Results All 11 responding centres reported having had to adapt or modify their services during the Spanish state of alarm (14 March–21 June 2020). One centre reported complete closure for 2 months and four reported increases in their operating hours. The average number of service users across all centres decreased by 22% in comparison to the same period in the previous year and the average needle distribution decreased by 40% in comparison to 2019. Most centres reported a decrease in infectious disease testing rates (hepatitis B and C viruses, human immunodeficiency virus, and tuberculosis) for March, April, and May in 2020 compared to the previous year. Reported deaths as a result of overdose did not increase during the state of alarm, but 2/11 (18%) centres reported an increase in overdose deaths immediately after finalisation of the state of alarm. Conclusion Overall, Spanish harm reduction centres were able to continue operating and offering services by adjusting operating hours. The number of overall service users and needles distributed fell during the Spanish state of alarm lockdown period, suggesting that fewer clients accessed harm reduction services during this time, putting them at greater risk of reusing or sharing injecting equipment, overdosing, acquiring infectious diseases with decreased access to testing or discontinuing ongoing treatment such as methadone maintenance therapy, hepatitis C treatment, or antiretroviral therapy.

Background Intracranial iatrogenic aneurysms (IIAs) developing after external ventricular drain (EVD) placement or removal have been only rarely reported. Most of these reports assumed a traumatic etiology; however, some have demonstrated an inflammatory origin. We have presented the case of an IIA that developed after an EVD had been inserted to treat acute hydrocephalus secondary to a ruptured arteriovenous malformation. We also performed a literature review and discussed how these IIAs might have an inflammatory rather than a traumatic etiology and how they might lie behind some of the cases of idiopathic hemorrhage observed after EVD manipulation. Case Description A 48-year-old woman had presented with acute hydrocephalus secondary to bleeding from a vermian arteriovenous malformation. The EVD inserted for hydrocephalus management required several revisions because of malfunction. Four weeks later, a diagnostic arteriogram had incidentally revealed the existence of a 5-mm aneurysm in an anterior branch of the right callosomarginal artery, coincidental to the EVD trajectory. The patient underwent emergent endovascular treatment of the aneurysm, with complete occlusion and no complications deriving from the procedure. Conclusions IIAs associated with EVD placement or removal have been described as a rare complication of the procedure. Despite the scarce number of cases reported and the lack of histological examinations, common features such as a distal location, an incidental diagnosis, and a benign clinical course might suggest a mycotic or inflammatory origin rather than a traumatic etiology. Considering the high volume of EVDs placed annually, this complication might be more frequent than reported. Further studies investigating the association of risk factors for inflammatory aneurysms and hemorrhage incidence after EVD could provide information on this intriguing topic.

The health effects of (poly)phenols (PPs) depend upon their bioavailability that, in general, is very low and shows a high interindividual variability. The low bioavailability of PPs is mainly attributed to their low absorption in the upper gastrointestinal tract as a result of their low water solubility, their presence in foods as polymers or in glycosylated forms, and their tight bond to food matrices. Although many studies have investigated how technological and biotechnological processes affect the phenolic composition of fruits and vegetables, limited information exists regarding their effects on PP bioavailability in humans. In the present review, the effect of food processing (mechanical, thermal, and non-thermal treatments), oral-delivery nanoformulations, enzymatic hydrolysis, fermentation, co-administration with probiotics, and generation of postbiotics in PP bioavailability have been overviewed, focusing in the evidence provided in humans., This research work was supported by the Projects 19900/GERM/15 (Fundación Séneca, Murcia, Spain), AGL2015-73107-EXP (MINECO,Spain), and CSIC 201870E014. Marta Pastor acknowledges a Juan de la Cierva Training Grant 2018 with Reference FJC2018-036570-I from the Ministry of Science and Innovation of the Spanish Government

A rapid analytical procedure is proposed for determining two antimicrobial onion organosulfur compounds, propyl disulfide (PDS) and propyl propane thiosulfonate (PTSO), in animal feed. The use of PTSO as a natural ingredient in animal feed is allowed due to its antimicrobial activity against pathogenic organisms. Two analytical methodologies using gas chromatography coupled to mass spectrometry (GC-MS) are compared. After the extraction of the compounds from animal feed with acetonitrile, dispersive solid phase extraction (DSPE) as a cleaning stage with C18, or dispersive liquid-liquid microextraction (DLLME), using 100 μL of CHCl3, was tried. Both the methods were validated using a pig feed sample and the best results were achieved by DLLME. This technique provided cleaner extracts, five-times greater linear ranges and lower detection limits than simple cleaning due to the enrichment factor achieved. The relative standard deviation decreased from 22% with DSPE to 13% with DLLME. The usefulness of the DLLME-GC-MS methodology was tested by analysing 10 different samples of chicken, calf, hen, cow and fish feed. The concentrations of PDS were in the 0.1-1.7 μg g-1 range and those of PTSO were between 0.09 and 2.1 μg g-1.

Background In settings with high antiretroviral therapy coverage, numerous health-related issues continue to undermine the health and health-related quality of life (HRQoL) of people living with HIV (PLHIV). As part of a larger study to develop and validate a new patient-reported outcome measure for use in HIV clinical care in Spain, we sought to identify the most burdensome health-related issues experienced by PLHIV in order to determine which issues should be addressed in the new instrument. Methods We conducted a literature review and a qualitative study based on four focus group discussions (FGDs) with key informants in Spain. Participants were selected via purposive sampling. Two FGDs convened 16 expert HIV service providers, and two convened 15 PLHIV with diverse epidemiological profiles. FGDs followed semi-structured interview scripts and incorporated an exercise to prioritise the most critical health-related issues among those named in the discussions. Content analysis was conducted using MAXQDA 12. Results The analysis of FGD data identified several broad categories of issues that were perceived to negatively affect PLHIV. The most frequently named issues fell within the categories of social problems; physical symptoms; psychological problems; and sexuality-related problems. Regarding social problems, stigma/discrimination was by far the issue raised the most frequently. In the prioritisation exercise, stigma/discrimination was also ranked as the most burdensome issue by both service providers and PLHIV. Within the physical symptoms category, the issues named most frequently were sleep-related problems, fatigue, physical pain and body fat changes. Regarding psychological problems, FGD participants most commonly spoke of emotional distress in general terms, and also called attention to depression and anxiety. In the prioritisation exercise, both service providers and PLHIV ranked psychological well-being as the second-most important issue following stigma. Sexuality-related problems that were reported included sexually transmitted infections, hormonal problems, lack of libido, and general sexual dissatisfaction. Conclusions PLHIV are negatively affected by a wide range of health-related issues. HIV-related stigma and psychological well-being remain major challenges. Identifying and addressing these and other issues in routine clinical care supports healthy aging and may ultimately contribute to better health and HRQoL outcomes in this population.

Hydrophilic interaction liquid chromatography using a zwitterionic phase, coupled with high-resolution mass spectrometry based on quadrupole and time-of-flight mass analyser (HILIC-Q-TOF-MS) is applied for the determination of four nucleobases (adenine, thymine, guanine and uracil), five nucleosides (adenosine, 5´-methyluridine, guanosine, cytidine and uridine) and five nucleotides (cytidine 5´-monophosphate, uridine 5´-monophosphate, adenosine 5´-monophosphate, inosine 5´-monophosphate and guanosine 5´-monophosphate) in food yeasts. Sample treatment based on the lysis of yeast using an ultra-turrax homogenizer allows the pellet and supernatant to be differentiated with low-speed centrifugation (1000 g). The pellet contains the nucleus (nuclear fraction), while the supernatant contains, among others, the organelles present in the cytoplasm (cytoplasmatic fraction). Both fractions were treated with perchloric acid in order to extract nucleobases, nucleosides and nucleotides and analysed by HILIC-Q-TOF-MS. The method was validated following international guidelines and detection limits were in the 2.5–22 ng mL−1 range (62–550 ng g−1, for 200 mg yeast). Five different Saccharomyces cerevisiae food yeasts were analysed and nucleotide concentrations of 0.6–570 µg g−1 and 5.6–924 µg g−1 were found in the nuclear and cytoplasmatic fractions of all samples. Nucleosides and nucleobases were not found in any sample.

Este artigo tem como objeto de estudo o processo de formação continuada para professores técnicos em um Centro de Educação Profissional do Semiárido, no Território de Identidade do Sisal, na Bahia, focalizado a partir de atitude orientada ao consenso, pelo viés de uma racionalidade comunicativa. Esse processo formativo é discutido com interfaces de saberes e práticas pedagógicas buscando contribuir para o desenvolvimento da profissionalidade docente do professor técnico, por meio da reflexão de concepções e de práticas pedagógicas. Assim, este artigo busca compreender como a formação continuada favorece o sobrepujamento do agir estratégico docente para um agir comunicativo na prática de ensino a fim de refletir sobre sua profissionalidade docente. A metodologia está embasada nos conceitos da pesquisa colaborativa, desenvolvida a partir de sessões reflexivas e de entrevistas semiestruturadas com professores técnicos sobre seu fazer docente e sua profissionalidade. Como resultado, para que esses técnicos tenham mais identificação com a docência, aponta-se a importância de processos formativos no espaço escolar, que surjam das próprias realidades educativas locais, de forma dialógica e engajada e contribuam para a ressignificação do trabalho do professor, no sentido de possibilitar

BackgroundNumerous health-related issues continue to undermine the health and health-related quality of life (HRQoL) of people living with HIV (PLHIV). We developed a clinic screening tool (CST-HIV) for the purpose of identifying these issues in routine specialist clinical care in Spain.MethodsWe used the following established instrument development procedures: (1) a literature review; (2) four focus group discussions (FGDs), two that convened 16 expert HIV care providers, and two that convened 15 PLHIV; (3) prioritisation, selection and definition of constructs (health-related issues) to include in the CST-HIV and drafting of initial item pool; and (4) a pilot study to analyse psychometric properties and validity of items and to determine which to retain in the final CST-HIV. The FGD interview scripts incorporated an exercise to prioritise the health-related issues perceived to have the greatest negative effect on HRQoL. The online questionnaire used for the pilot study included the pool of CST-HIV items and validated measures of each construct.ResultsWe identified 68 articles that reported on factors associated with the HRQoL of PLHIV. The most burdensome health-related issues identified in the FGDs related to stigma, socioeconomic vulnerability, sleep/fatigue, pain, body changes, emotional distress, and sexuality. Based on the literature review and FGD findings, we selected and defined the following constructs to include in the initial CST-HIV: anticipated stigma, emotional distress, sexuality, social support, material deprivation, sleep/fatigue, cognitive problems, and physical symptoms. Two researchers wrote six to eight items for each construct. Next, 18 experts rated 47 items based on their clarity, relevance, and representativeness. Pilot testing was carried out with 226 PLHIV in Spain. We retained 24 items based on empirical criteria that showed adequate psychometric properties. Confirmatory factor analysis confirmed the eight-factor structure with a good fit to the data (RMSEA = 0.035, AGFI = 0.97, CFI = 0.99). We found strong positive correlations between the instrument’s eight dimensions and validated measures of the same constructs. Likewise, we found negative associations between the dimensions of the CST-HIV and HRQoL.ConclusionThe CST-HIV is a promising tool for use in routine clinical care to efficiently identify and address health-related issues undermining the HRQoL of PLHIV.

The presence of benzene, toluene, ethylbenzene and xylenes (BTEX) in dyes, paints and pigments is not still regulated but might be an indicator of contamination due to the well-known health risk of these compounds. An analytical procedure based on headspace sorptive extraction using a polydimethylsiloxane stir bar, was developed for the gas chromatographic determination of BTEX in finger paint samples. Univariate and multivariate designs were used to optimize experimental parameters, including equilibrium time, desorption time, saline effect and sample mass. Thirty minutes were necessary to absorb the analytes in the polydimethylsiloxane stir bar followed by a 5 min thermal desorption step. Limits of quantification of between 0.036 and 0.12 ng g−1 depending on the compound, were obtained. BTEX were found in the finger paint samples at levels between 0.14 and 12 ng g−1.

The photocatalytic efficiency of magnetic nanoparticles using different coatings between Fe3O4 and semiconductor oxides (ZnO and TiO2) was evaluated by their capability to degrade four insecticides (etoxazole, fenpyroximate, flonicamid and metaflumizone) in water exposed to sunlight. Magnetic particles coated with SiO2 (MSPs) showed higher photodegradation activity. Sol-gel TiO2-SiO2 coated TiO2 P25 magnetic particles (MSTi2Ps) and sol-gel ZnO-SiO2 coated ZnO magnetic particles (MSZn2Ps) were more efficient than magnetic particles consisting of a silica undercoat followed by a TiO2- or ZnO- sol-gel coating as the photocatalytic layer for catalysing the removal of these pesticides. Finally, the addition of Na2S2O8 increased the photooxidation activity of both MSTi2Ps and MSZn2Ps. A comparison of the semiconductor oxides showed MSZn2Ps to be the most effective for the removal of the studied pollutants. As a consequence, the tandem MSZn2Ps/Na2S2O8 provides an interesting combination to remove pesticides in water.

Highly active transmembrane anion transporters have demonstrated their activity against antibiotic-resistant and clinically relevant bacterial strains. This type of compound offers promise as a strategy to develop novel antibacterial agents.

Background There is little verified information on global healthcare utilization by irregular migrants. Understanding how immigrants use healthcare services based on their needs is crucial to establish effective health policy. We compared healthcare utilization between irregular migrants, documented migrants, and Spanish nationals in a Spanish autonomous community. Methods This retrospective, observational study included the total adult population of Aragon, Spain: 930,131 Spanish nationals; 123,432 documented migrants; and 17,152 irregular migrants. Healthcare utilization data were compared between irregular migrants, documented migrants and Spanish nationals for the year 2011. Multivariable standard or zero-inflated negative binomial regression models were generated, adjusting for age, sex, length of stay, and morbidity burden. Results The average annual use of healthcare services was lower for irregular migrants than for documented migrants and Spanish nationals at all levels of care analyzed: primary care (0.5 vs 4 vs 6.7 visits); specialized care (0.2 vs 1.8 vs 2.9 visits); planned hospital admissions (0.3 vs 2 vs 4.23 per 100 individuals), unplanned hospital admissions (0.5 vs 3.5 vs 5.2 per 100 individuals), and emergency room visits (0.4 vs 2.8 vs 2.8 per 10 individuals). The average annual prescription drug expenditure was also lower for irregular migrants (€9) than for documented migrants (€77) and Spanish nationals (€367). These differences were only partially attenuated after adjusting for age, sex, and morbidity burden. Conclusions Under conditions of equal access, healthcare utilization is much lower among irregular migrants than Spanish nationals (and lower than that of documented migrants), regardless of country of origin or length of stay in Spain.

Background: Containment policies and other restrictions introduced by the Spanish government in response to the COVID-19 pandemic present challenges for marginalised populations, such as people who use drugs. Harm reduction centres are often linked to social services, mental health services, and infectious disease testing, in addition to tools and services that help to reduce the harms associated with injecting drugs. This study aimed to explore the impact of the pandemic on these services in four autonomous communities in Spain. Methods: This is a cross-sectional study that employed a seven-section structured survey administered electronically to 20 centres in July 2020. Data from the most heavily affected months (March–June) in 2020 were compared to data from the same period in 2019. Averages were calculated with their ranges, rates, and absolute numbers.Results: All 11 responding centres reported having had to adapt or modify their services during the Spanish state of alarm (14 March–21 June 2020). One centre reported complete closure for two months and four reported increases in their operating hours. The average number of service users across all centres decreased by 22% in comparison to the same period in the previous year and the average needle distribution decreased by 40% in comparison to 2019. Most centres reported a decrease in infectious disease testing rates (hepatitis B and C viruses [HBV, HCV], human immunodeficiency virus (HIV), and tuberculosis [TB]) for March, April, and May in 2020 compared to the previous year. Reported deaths as a result of overdose did not increase during the state of alarm, but 2/11 (18%) centres reported an increase in overdose deaths immediately after finalisation of the state of alarm.Conclusion: Overall, Spanish harm reduction centres were able to continue operating and offering services by adjusting operating hours. The number of overall service users and needles distributed fell during the Spanish state of alarm lockdown period, suggesting that fewer clients accessed harm reduction services during this time, putting them at greater risk of reusing or sharing injecting equipment, overdosing, acquiring infectious diseases with decreased access to testing or discontinuing ongoing treatment such as methadone maintenance therapy, hepatitis C treatment, or antiretroviral therapy.

Background: Containment policies and other restrictions introduced by the Spanish government in response to the COVID-19 pandemic present challenges for marginalised populations, such as people who use drugs. Harm reduction centres are often linked to social services, mental health services, and infectious disease testing, in addition to tools and services that help to reduce the harms associated with injecting drugs. This study aimed to explore the impact of the pandemic on these services in four autonomous communities in Spain.Methods: This is a cross-sectional study that employed a seven-section structured survey administered electronically to 20 centres in July 2020. Data from the most heavily affected months (March–June) in 2020 were compared to data from the same period in 2019. Averages were calculated with their ranges, rates, and absolute numbers.Results: All 11 responding centres reported having had to adapt or modify their services during the Spanish state of alarm (14 March–21 June 2020). One centre reported complete closure for two months and four reported increases in their operating hours. The average number of service users across all centres decreased by 22% in comparison to the same period in the previous year and the average needle distribution decreased by 40% in comparison to 2019. Most centres reported a decrease in infectious disease testing rates (hepatitis B and C viruses [HBV, HCV], human immunodeficiency virus (HIV), and tuberculosis [TB]) for March, April, and May in 2020 compared to the previous year. Reported deaths as a result of overdose did not increase during the state of alarm, but 2/11 (18%) centres reported an increase in overdose deaths immediately after finalisation of the state of alarm.Conclusion: Overall, like other European countries, Spanish harm reduction centres were able to continue operating and offering services by adjusting operating hours. The number of overall service users and needles distributed fell during the Spanish state of alarm lockdown period, suggesting that fewer clients accessed harm reduction services during this time, putting them at greater risk of reusing or sharing injecting equipment, overdosing, acquiring infectious diseases with decreased access to testing or discontinuing ongoing treatment (methadone maintenance therapy, hepatitis C treatment, or antiretroviral therapy).

Background: This study was to describe the evolution of patients who underwent surgical treatment of drug- resistant occipital lobe epilepsy (OLE) at our institution. Methods: We performed a retrospective analysis of data collected from electronic and paper clinical records of 20 patients who were diagnosed of OLE and underwent epilepsy surgery at our institution between 1998 and 2018. We also contacted patients by telephone and asked them to fill out a questionnaire about quality of life in epilepsy (QOLIE-10). Assembled data were analyzed using descriptive statistics. Results: The age at surgery ranged between 19 and 55 years. The period encompassing epilepsy onset and the date of surgery was variable. Semiology of seizures included visual symptoms in 75% of patients. In 90% of cases subdural grids, depth electrodes or a combination of both were used to plan the surgery. The most frequent neuroimaging and histopathological finding was cortical dysplasia (55%). The postoperative follow-up period was up to 15 years. The most common score on the Engel scale was I (70%). Visual deficits increased after surgery. Median score on QOLIE-10 questionnaire was 82.5 (interquartile range: 32.5). Conclusion: Surgical treatment of drug-resistant OLE offers hopeful results to those patients who have run out of pharmacological options and leads to postoperative deficits that are deemed expectable and occasionally acceptable.

Background There is little verified information on the global health status of undocumented migrants (UMs). Our aim is to compare the prevalence of the main chronic diseases and of multimorbidity in undocumented migrants, documented migrants, and Spanish nationals in a Spanish autonomous community. Methods Retrospective observational study of all users of the public health system of the region of Aragon over 1 year (2011): 930,131 Spanish nationals; 123,432 documented migrants (DMs); and 17,152 UMs. Binary logistic regression was performed to examine the association between migrant status (Spanish nationals versus DMs and UMs) and both multimorbidity and individual chronic diseases, adjusting for age and sex. Results The prevalence of individual chronic diseases in UMs was lower than in DMs and much lower than in Spanish nationals. Comparison with the corresponding group of Spanish nationals revealed odds ratios (OR) of 0.1–0.3 and 0.3–0.5 for male and female UMs, respectively (p

This work focuses on the development, validation and application of an analytical method for the determination of twenty organochlorine pesticides (OCPs) in human tissues using salting-out liquid–liquid extraction and dispersive liquid–liquid microextraction for sample preparation and gas chromatography–mass spectrometry to analyze the obtained extracts. Measurement of the concentration levels of these toxics in tissues can be used to assess the risk of the population to exposure. The linearity of the proposed method was verified in the 10–1,000 ng/g range. The sensitivity was evaluated calculating the limits of detection (LODs) for 20 OCPs (α-, β-, γ- and δ-hexachlorocyclohexane (HCH), α- and β-endosulfan, endosulfan sulfate, aldrin, dieldrin, endrin, endrin ketone, endrin aldehyde, α- and γ-chlordane, 4,4′-dichlorodiphenyltrichloroethane, 4,4′-dichlorodiphenyldichloroethylene (DDE), 4,4′-dichlorodiphenyldichloroethane, heptachlor, heptachlor epoxide and methoxychlor), most of them being found between 1.0 and 16 ng/g. The intra- and interday precisions were

This study describes the use of a magnetic multi-walled carbon nanotube composite for the preconcentration of nine parabens (methyl, ethyl, propyl, isopropyl, butyl, isobutyl, pentyl, phenyl and benzylparaben) from water and urine samples after in situ acetylation. The enriched extracts were obtained by dispersive magnetic solid phase extraction (DMSPE) and analysed by gas chromatography with mass spectrometry (GC-MS). Several parameters affecting the adsorption (extraction time, magnetic material mass and ionic strength) and desorption (desorption solvent nature and volume, and desorption time) DMSPE steps were investigated. Matrix-matched calibration was recommended for quantification of the samples. Linearities in the 0.5-150 ng mL-1 range were obtained, depending on the compound. Under the optimal experimental conditions, the limits of detection ranged between 0.03 and 2.0 ng mL-1, depending on the sample matrix and the paraben congener. None of the samples analysed contained the above mentioned parabens. The method was validated with two water samples (harbour and wastewater treatment plant) and two urine samples spiked at two concentration levels. Recoveries in the 81-119% range were obtained.

The determination of 15 pyrethroids in soil and water samples was carried out by gas chromatography with mass spectrometry. Compounds were extracted from the soil samples (4 g) using solid-liquid extraction and then salting-out assisted liquid-liquid extraction. The acetonitrile phase obtained (0.8 mL) was used as a dispersant solvent, to which 75 μL of chloroform was added as an extractant solvent, submitting the mixture to dispersive liquid-liquid microextraction. For the analysis of water samples (40 mL), magnetic solid-phase extraction was performed using nanocomposites of magnetic nanoparticles and multiwalled carbon nanotubes as sorbent material (10 mg). The mixture was shaken for 45 min at room temperature before separation with a magnet and desorption with 3 mL of acetone using ultrasounds for 5 min. The solvent was evaporated and reconstituted with 100 μL acetonitrile before injection. Matrix-matched calibration is recommended for quantification of soil samples, while water samples can be quantified by standards calibration. The limits of detection were in the range of 0.03-0.5 ng/g (soil) and 0.09-0.24 ng/mL (water), depending on the analyte. The analyzed environmental samples did not contain the studied pyrethroids, at least above the corresponding limits of detection.

ZnO (zincite) coated magnetic particles were successfully prepared and characterized by different techniques (XRD, FE-SEM, EDX, and FTIR). The photocatalytic efficiency of the magnetic particles was assessed by their ability to degrade six fungicides in water exposed to solar irradiation. Magnetic particles composed of a silica undercoat beneath a ZnO coating (MSZPs) were more efficient than magnetic particles loaded with ZnO alone (MZPs) for catalysing the removal of these fungicides. The optimum mass ratio of SiO 2 to ZnO was about 1:2 because of almost complete coverage by ZnO surface coverage and the high photocatalytic degradation rates obtained for the studied fungicides, which was comparable to that obtained with ZnO nanopowder. However, in contrast to ZnO nanopowder, the photodissolution of Zn using MSZPs was low, with Zn 2+ concentration at the end of the experiment (0.75 mg L −1 ) below legally permitted levels in drinking water. In addition, the effect of initial concentration of fungicides was studied in order to know maximum degradation efficiency. Finally, the photodegradation activity of MSZPs was increased by the addition of Na 2 S 2 O 8 (250 mg L −1 ). Therefore, the tandem MSZPs/Na 2 S 2 O 8 provides an interesting method to degrade some organic pollutants in water.

Nitrosamines (NAs), which are catalogued as carcinogenic compounds, may be present in meat products due to the conversion of nitrites and as result of migration from elastic rubber nettings used. A method based on ultrasonic assisted extraction coupled with dispersive liquid–liquid microextraction as sample treatment and gas chromatography-mass spectrometry as separation and detection technique was proposed for the determination of twelve NAs in cooked ham samples. The method was validated by evaluating linearity (0.5–1000 ng g−1), matrix effect, sensitivity (detection limits were between 0.15 and 1.4 ng g−1) and precision, which was below 12%. Five NAs were found in the samples with levels ranging from not quantifiable to 40 ng g−1. The effect of the elastic rubber nettings on the nitrosamine content of meat was evaluated by comparing the levels found in products made with several plastics or thread in the presence of additives.

Authentication of organic olives and olive oils is of growing interest due to the marked development of organic olive agriculture worldwide. In this study, the potential of using the untargeted volatile fingerprinting of organic and conventional olives obtained with a low-cost and fast methodology based on headspace ultraviolet ion mobility spectrometry (HS-UV-IMS) was investigated for first time. The results showed the suitability of the optimised method since a 94.39% success rate of classification of olives samples was achieved. However, this HS-UV-IMS device did not provide good results for olive oil samples and so two different and more powerful analytical techniques (HS-gas chromatography coupled to IMS with tritium source (HS-GC-IMS) and capillary electrophoresis coupled to UV detector (CE-UV)) were employed respectively to analyse the volatile fingerprinting and the polar compounds extracted from organic and conventional olive oils from two varieties and harvests. It was found that both devices could provide good classification results, above 93.20%, if an adequate number of samples (n > 50) from a single harvest was used. However, it was demonstrated that CE-UV is the best option since it provided more robust classification results over years (95.76%) in comparison with HS-GC-IMS (75.16%). An improvement in classification percentages of HS-GC-IMS data was observed when samples from two harvests were separated into Picual (98.73%) and Hojiblanca (98.78%) variety sets. HS-GC-mass spectrometry with a single quadruple (HS-GC-MS) was employed to contrast the results obtained for the volatile fraction of olive and olive oils with HS-UV-IMS and HS-GC-IMS devices. Also, up to 12 compounds were identified in olive and olive oil samples using HS-GC-MS but only the 3-hexenyl acetate was able to differentiate organic and conventional olive oils.

A multiresidue method was developed to determine twenty-five fungicides belonging to three different chemical families, oxazoles, strobilurins and triazoles, in water and fruit samples, using dispersive liquid-liquid microextraction (DLLME) and liquid chromatography/tandem mass spectrometry (LC-MS2). Solid-liquid extraction with acetonitrile was used for the analysis in fruits, the extract being used as dispersant solvent in DLLME. Since some of the analytes showed high affinity for chloroform and the others were more efficiently extracted with undecanol, a mixture of both solvents was used as extractant in DLLME. After evaporation of CHCl3, the enriched phase was analyzed. Enrichment factors in the 23-119 and 12-60 ranges were obtained for waters and fruits, respectively. The approach was most sensitive for metominostrobin with limits of quantification of 1ngL-1 and 5ngkg-1 in waters and fruits, respectively, while a similar sensitivity was attained for tebuconazole in fruits. Recoveries of the fungicides varied between 86 and 116%.

Magnetic solid phase extraction (MSPE) with cobalt ferrite nanoparticles coated with oleic acid is described for the determination of alkylphenols (APs), 4-tert-butylphenol (TBP), 4-pentylphenol (PP), 4-hexylphenol (HP), 4-tert-octylphenol (TOP), 4-n-octylphenol (OP) and 4-nonylphenol (NP) in baby foods using gas chromatography with mass spectrometry (GC–MS). Prior to MSPE, the sample was treated with trichloroacetic acid, and the APs derivatized with acetic anhydride. Parameters affecting the extraction efficiency: amount of magnetic nanoparticles, extraction time and desorption conditions, were optimized. The enriched phase obtained was evaporated to dryness and the residue reconstituted in 50 μL of methanol, 1 μL of which was injected into the GC–MS. Samples were quantified applying matrix-matched calibration and using 2-chloro-5-bromoanisole as surrogate standard. The analysis of 0.5 g of sample provided detection limits in the 0.4–1.7 ng g −1 range. Some samples contained APs at levels of between 3 ng g −1 for HP and 122 ng g −1 for TOP.

An analytical procedure is proposed for determining three cyanotoxins (microcystin RR, microcystin LR, and nodularin) and two phycotoxins (domoic and okadaic acids) in seawater and algae-based food supplements. The toxins were first isolated by a salting out liquid extraction procedure. Since the concentration expected in the samples was very low, a dispersive liquid&ndash, liquid microextraction procedure was included for preconcentration. The ionic liquid 1-hexyl-3-methylimidazolium hexafluorophosphate (80 mg) was used as green extractant solvent and acetonitrile as disperser solvent (0.5 mL) for a 10 mL sample volume at pH 1.5, following the principles of green analytical chemistry. Liquid chromatography with electrospray ionization and quadrupole time of flight-mass spectrometry (LC-Q-TOF-MS) was used. The selectivity of the detection system, based on accurate mass measurements, allowed the toxins to be unequivocally identified. Mass spectra for quadrupole time of flight-mass spectrometry (Q-TOF-MS) and Q-TOF-MS/MS were recorded in the positive ion mode and quantification was based on the protonated molecule. Retention times ranged between 6.2 and 17.9 min using a mobile phase composed by a mixture of methanol and formic acid (0.1%). None of the target toxins were detected in any of the seawater samples analyzed, above their corresponding detection limits. However, microcystin LR was detected in the blue green alga sample.

Polycyclic aromatic hydrocarbons (PAHs) are considered xenobiotics of a potentially carcinogenic nature, being accumulated in the fatty tissue of the body. The objective of this work was the development and validation of a new analytical method to check the bioaccumulation of these toxic compounds in human organs obtained from autopsies. The contaminants were first isolated from the tissues by salt-assisted liquid-liquid extraction in acetonitrile. Because of the low concentrations of these compounds in the human body, a dispersive liquid-liquid microextraction procedure was included. The preconcentrated samples were analyzed by gas chromatography-mass spectrometry to identify the compounds. Principal component analysis was applied to show the natural clustering of forensic samples and orthogonal partial least-squares discriminant analysis to develop a multivariate regression method, which permitted the classification of samples. The quantification limits for the 13 PAHs (acenaphthylene, fluorene, phenanthrene, anthracene, pyrene, benzo(a)anthracene, chrysene, benzo(b)fluoranthene, benzo(k)fluoranthene, benzo(a)pyrene, dibenz(a,h)anthracene, benzo(g,h,i)perylene, and indeno(1,2,3-cd)pyrene) analyzed were in the 0.06-0.44 ng g-1 range, depending on the compound, while the mean intraday relative standard deviation of about 7% demonstrated the high precision of the method. Linearity was verified in the 0.5-200 ng g-1 range, and the enrichment factors were between 55 and 122. The results provided by the analysis of seven different human organs (brain, liver, kidney, lung, heart, spleen, and abdominal fat) from eight autopsies confirmed the PAH-bioaccumulation capacity of human body, fat showing the highest degree of bioaccumulation. The present work is the first study on PAH contamination in different organs obtained from autopsies, being PAH detected in most human samples at values ranged from 0 to 19 ng g-1.

Iron is essential for most life forms. Under iron-limiting conditions, many bacteria produce and release siderophores—molecules with high affinity for iron—which are then transported into the cell in their iron-bound form, allowing incorporation of the metal into a wide range of cellular processes. However, free iron can also be a source of reactive oxygen species that cause DNA, protein, and lipid damage. Not surprisingly, iron capture is finely regulated and linked to oxidative-stress responses. Here, we provide evidence indicating that in the plant-beneficial bacterium Pseudomonas putida KT2440, the amino acid l-arginine is a metabolic connector between iron capture and oxidative stress. Mutants defective in arginine biosynthesis show reduced production and release of the siderophore pyoverdine and altered expression of certain pyoverdine-related genes, resulting in higher sensitivity to iron limitation. Although the amino acid is not part of the siderophore side chain, addition of exogenous l-arginine restores pyoverdine release in the mutants, and increased pyoverdine production is observed in the presence of polyamines (agmatine and spermidine), of which arginine is a precursor. Spermidine also has a protective role against hydrogen peroxide in P. putida, whereas defects in arginine and pyoverdine synthesis result in increased production of reactive oxygen species. IMPORTANCE The results of this study show a previously unidentified connection between arginine metabolism, siderophore turnover, and oxidative stress in Pseudomonas putida. Although the precise molecular mechanisms involved have yet to be characterized in full detail, our data are consistent with a model in which arginine biosynthesis and the derived pathway leading to polyamine production function as a homeostasis mechanism that helps maintain the balance between iron uptake and oxidative-stress response systems., This work was funded by grants BFU2016-80122-P from the Plan Estatal de I+D+I and P11-CVI-7391 from Junta de Andalucía and by EFDR funds. L.B.-M. was the recipient of a predoctoral contract from Junta de Andalucía.

Re-activation of the healing process is a major challenge in the field of chronic wound treatment. For that purpose, lipid-nanoparticles, especially nanostructured lipid carriers (NLC), possess extremely useful characteristics such as biodegradability, biocompatibility and long-term stability, besides being suitable for drug delivery. Moreover, they maintain wound moisture due to their occlusive properties, which have been associated with increased healing rates. In the light of above, NLC have been extensively used topically for wound healing; but to date, there are no safety-preclinical studies concerning such type of application. Thus, in this work, biodistribution studies were performed in rats with the NLC previously developed by our research group, using technetium-99 m (99mTc-NLC) as radiomarker, topically administered on a wound. 99mTc-NLC remained on the wound for 24 h and systemic absorption was not observed after administration. In addition, toxicological studies were performed to assess NLC safety after topical administration. The results obtained demonstrated that NLC were non-cytotoxic, non-sensitizing and non-irritant/corrosive. Overall, it might be concluded that developed NLC remained at the administration area, potentially exerting a local effect, and were safe after topical administration on wounds.

OBJECTIVEThe percentage of women publishing high-impact neurosurgical research might be perceived as a representation of our specialty and may influence the perpetuation of the existing gender gap. This study investigated whether the trend in women taking lead roles in neurosurgical research has mirrored the increase in female neurosurgeons during the past decade and whether our most prestigious publications portray enough female role models to stimulate gender diversity among the new generation of neurosurgeons.METHODSTwo of the most prominent neurosurgical journals—Journal of Neurosurgery and Neurosurgery—were selected for this study, and every original article that was published in 2009 and 2019 in each of those journals was investigated according to the gender of the first and senior authors, their academic titles, their affiliations, and their institutions’ region.RESULTSA total of 1328 articles were analyzed. The percentage of female authors was significantly higher in Europe and Russia compared with the US and Canada (first authors: 60/302 [19.9%] vs 109/829 [13.1%], p = 0.005; and senior authors: 32/302 [10.6%] vs 57/829 [6.9%], p = 0.040). Significantly increased female authorship was observed from 2009 to 2019, and overall numbers of both first and senior female authors almost doubled. However, when analyzing by regions, female authorship increased significantly only in the US and Canada. Female authors of neurosurgical research articles were significantly less likely to hold an MD degree compared with men. Female neurosurgeons serving as senior authors were represented in only 3.6% (48/1328) of articles. Women serving as senior authors were more likely to have a female colleague listed as the first author of their research (29/97 [29.9%] vs 155/1231 [12.6%]; χ2 = 22.561, p = 0.001).CONCLUSIONSAlthough this work showed an encouraging increase in the number of women publishing high-impact neurosurgical research, the stagnant trend in Europe may suggest that a glass ceiling has been reached and further advances in equity would require more aggressive measures. The differences in the researchers’ profiles (academic title and affiliation) suggest an even wider gender gap. Cultural unconscious bias may explain why female senior authors have more than double the number of women serving as their junior authors compared with men. While changes in the workforce happen, strategies such as publishing specific issues on women, encouraging female editorials, and working toward more gender-balanced editorial boards may help our journals to portray a more equitable specialty that would not discourage bright female candidates.

Ranitidine drug products were recently recalled because they contained carcinogenic nitrosamines (NAs), such as N-nitrosodimethylamine (NDMA) and N-nitrosodiethylamine (NDEA). This episode emphasises the importance of developing analytical methods to determine NAs in this type of product. This study describes the development and validation of an analytical method for the determination of nine NAs (NDMA, N-nitrosomethylethylamine (NEMA), NDEA, N-nitrosopyrrolidine (NPYR), N-nitrosomorpholine (NMOR) N-nitrosodi-n-propylamine (NDPA) N-nitrosopiperidine (NPIP), N-nitrosodi-n-butylamine (NDBA) and N-nitrosodiphenylamine (NDPhA)) in ranitidine drug samples using a combination of microextraction and gas chromatography-mass spectrometry. The procedure involved the dissolution of 1 g of sample in 10 mL of water. For the dispersive liquid-liquid microextraction, 0.5 g of NaCl was added to this aqueous solution, followed by a mixture containing 0.5 mL methanol as dispersant and 150 μL chloroform as extractant solvent. The recovered organic phase was injected into the GC-MS system and a 20 min oven programme was applied. Quantification limits were in the 0.21–21 ng g−1 range, corresponding the lower limit to NDPhA and the higher to NDMA. Relative standard deviations lower than 12% were achieved in all cases, which indicates the adequate precision of the method. Seven pharmaceutical products containing two different amounts of ranitidine (150 and 300 mg) were analysed. None of the samples contained NEMA, NDEA, NPYR, NMOR, NDPA or NPIP, while NDMA, NDBA and NDPhA were found in three products.

In the last decades, the encapsulation of antibiotics into nanoparticulate carriers has gained increasing attention for the treatment of infectious diseases. Sodium colistimethate-loaded solid lipid nanoparticles (Colist-SLNs) and nanostructured lipid carriers (Colist-NLCs) were designed aiming to treat the pulmonary infection associated to cystic fibrosis patients. The nanoparticles were freeze-dried using trehalose as cryoprotectant. The stability of both nanoparticles was analysed over one year according to the International Conference of Harmonisation (ICH) guidelines by determining the minimum inhibitory concentration (MIC) against clinically isolated Pseudomonas aeruginosa strains and by studying their physico-chemical characteristics. The results showed that Colist-SLNs lost their antimicrobial activity at the third month; on the contrary, the antibacterial activity of Colist-NLCs was maintained throughout the study within an adequate range (MIC ≤16 μg/mL). In addition, Colist-NLCs exhibited suitable physico-chemical properties at 5 °C and 25 °C/60% relative humidity over one year. Altogether, Colist-NLCs proved to have better stability than Colist-SLNs.

A rapid and simple procedure is reported for the determination of six ethylene glycol ethers in cleaning products and detergents using gas chromatography with mass spectrometry. The analytes were extracted from 2.0 g samples in acetonitrile (3 mL) and the extract was submitted to a clean-up step by QuEChERS method, using a mixture containing 0.3 g magnesium sulfate, 0.15 g primary/secondary amine, and 0.05 g C18 . The clean acetonitrile extract (1 μL) was injected into the chromatographic system. No matrix effect was observed, so the quantification of the samples was carried out against external standards. Detection limits were in the range 3.0-27 ng/g for the six ethylene glycol ethers. The recoveries obtained, using the optimized procedure, were in the 89.4-118% range, with relative standard deviations lower than 14%. Twenty-three different household cleaning products, including glass cleaner, degreaser, floor, softeners, and clothes and dishwashing detergents, were analyzed. Large interindividual variations were observed between samples and compounds.

Magnetic nanoparticles of cobalt ferrite with oleic acid as the surfactant (CoFe2O4/oleic acid) were used as sorbent material for the determination of alkylphenols in fruit juices. High sensitivity and specificity were achieved by liquid chromatography and detection using both diode-array (DAD) and electrospray-ion trap-tandem mass spectrometry (ESI-IT-MS/MS) in the selected reaction monitoring (SRM) mode of the negative fragment ions for alkylphenols (APs) and in positive mode for ethoxylate APs (APEOs). The optimized conditions for the different variables influencing the magnetic separation procedure were: mass of magnetic nanoparticles, 50mg, juice volume, 10mL diluted to 25mL with water, pH 6, stirring for 10min at room temperature, separation with an external neodymium magnet, desorption with 3mL of methanol and orbital shaking for 5min. The enriched organic phase was evaporated and reconstituted with 100µL acetonitrile before injecting 30µL into a liquid chromatograph with a mobile phase composed of acetonitrile/0.1% (v/v) formic acid under gradient elution. Quantification limits were in the range 3.6 to 125ngmL(-1). The recoveries obtained were in the 91-119% range, with RSDs lower than 14%. The ESI-MS/MS spectra permitted the correct identification of both APs and APEOs in the fruit juice samples.

Among the pathogens that affect cystic fibrosis (CF) patients, Pseudomonas aeruginosa is the most prevalent. As a way to fight against this infection, nanotechnology has emerged over the last decades as a promising alternative to overcome resistance to antibiotics in infectious diseases. The goal of this work was to elaborate and characterize lipid nanoparticles for pulmonary delivery of tobramycin. Tobramycin-loaded nanostructured lipid carriers (Tb-NLCs) were prepared by hot melt homogenization technique. In addition, nanoparticles labeled with infrared dye (IR-NLCs) were used to investigate their in vivo performance after pulmonary administration. Tb-NLCs displayed a mean diameter size around 250 nm, high drug encapsulation (93%) and sustained release profile. Tb-NLCs showed to be active against clinically isolated P. aeruginosa. Moreover, Tb-NLCs did not decrease cell viability and were able to overcome an artificial mucus barrier in the presence of mucolytics agents. During the in vivo assay, IR-NLCs were administered to several mice by the intratracheal route using a Penn Century device. Next, the biodistribution of the nanoparticles was analyzed at different time points showing a wide nanosystem distribution in the lungs. Altogether, tobramycin-loaded NLCs seem to us an encouraging alternative to the currently available CF therapies.

Sodium colistimethate (SCM) and amikacin (AMK) are among the few antibiotics effective against resistant P. aeruginosa, K. pneumoniae and A. baumannii; however, their toxicity severely limits their use. Enclosing antibiotics into nanostructured lipid carriers (NLC) might decrease drug toxicity and improve antibiotic disposition. In this work, SCM or AMK were loaded into different NLC formulations, through high pressure homogenization, and their in vitro and in vivo effectiveness was analyzed. The encapsulation process did not reduce drug effectiveness since in vitro SCM-NLC and AMK-NLC drug activity was equal to that of the free drugs. As cryoprotectant, trehalose showed better properties than dextran. Instead, positive chitosan coating was discarded due to its limited cost-efficiency. Finally, the in vivo study in acute pneumonia model revealed that intraperitoneal administration was superior to the intramuscular route and confirmed that (−) SCM-NLC with trehalose, was the most suitable formulation against an extensively drug-resistant A. baumannii strain.

A rapid procedure for the determination of amphenicol antibiotics in human urine by liquid chromatography with quadrupole time-of-flight mass spectrometry (LC-QTOF-MS) is proposed. The presence of thiamphenicol (TAP), florfenicol (FF) and chloramphenicol (CAP) in the human body can be attributed to their administration to treat certain diseases or by eating food of animal origin. The TAP, FF and CAP excreted in urine is mainly in the form of glucuronide conjugates, although their free forms may also be excreted to a lesser extent. In the procedure described, the enzymatic hydrolysis of amphenicol glucuronide forms in urine was carried out using β-glucuronidase and sulfatase at pH 5 (37 °C, overnight) in order to discriminate the free and conjugated forms. Then, amphenicol antibiotics were submitted to dispersive liquid–liquid microextraction (DLLME) for preconcentration. All the parameters affecting DLLME efficiency were optimized, and the following conditions were selected: 0.9 g NaCl in 10 mL of urine, to which 1.2 mL methanol (as dispersant solvent) and 1 mL of 4-methyl-2-pentanone (as extractant solvent) were added. The absence of a matrix effect allowed quantification of the samples against aqueous standards. Detection limits were 29, 6 and 3 pg mL−1 for TAP, FF and CAP, respectively. Relative standard deviations were calculated to evaluate the intra- and inter-day precision and values lower than 10% were obtained in all cases. The trueness of the method was tested through recovery studies, obtaining satisfactory values (83–104%). Ten urine samples obtained from volunteers were analysed and all of them were free of the studied antibiotics.

The validation of a procedure for the determination of six alkylphenols (APs), 4-tert-butylphenol, 4-pentylphenol, 4-tert-octylphenol, 4-hexylphenol, 4-octylphenol and 4-nonylphenol, and three bisphenols (BPs), bisphenol A, bisphenol F and bisphenol Z, in seven human organs and tissues (kidney, liver, heart, lung, spleen, brain and abdominal fat) obtained from eight autopsies is presented. Previously ground samples were treated by salt-assisted liquid-liquid extraction (SALLE) for isolation of the analytes and then pre-concentrated using dual stir bar sorptive extraction (SBSE), allowing two different extraction conditions for the same sample. Finally, thermal desorption was used as the injector system in combination with gas chromatography coupled to mass spectrometry (GC-MS). To determine BPs, derivatization using acetic anhydride was required, although this step was not necessary for the APs. Two parallel extractions of the contaminants with the stir bars were performed, followed by thermal desorption and chromatographic analysis. The procedure provided quantification limits between 0.050 and 4.0 ng g−1 for APs and from 0.26 to 2.6 ng g−1 for BPs. Repeatability and reproducibility values were lower than 15% in all cases. The accuracy of the procedure was established by a recovery study, which provided values in the 85.8–115% range for APs and 83.6–120% for BPs. Samples were analyzed with the proposed methodology and data were processed by ANOVA tests to study the behaviour and bioaccumulation of these compounds in human tissues or organs. In addition, discriminant analysis detected age- and sex-dependent differences in bioaccumulation.

The usefulness of nanotechnology to increase the bioavailability of drugs and decrease their toxicity may be a tool to deal with multiresistant P. aeruginosa (Mr-Pa) respiratory infections. We describe the preparation and the in vivo efficacy and safety of sodium colistimethate-loaded nanostructured lipid carriers (SCM-NLC) by the pulmonary and intramuscular routes. Nanoparticles showed 1-2 mg/L minimum inhibitory concentration against eight extensively drug-resistant P. aeruginosa strains. In vivo, SCM-NLC displayed significantly lower CFU/g lung than the saline and similar to that of the free SCM, even the dose in SCM-NLC group was lower than free SCM. There was no tissue damage related to the treatments. Biodistribution assessments showed a mild systemic absorption after nebulization and a notorious absorption after IM route. Altogether, it could be concluded that SCM-NLC were effective against P. aeruginosa in vivo, not toxic and distribute efficiently to the lung and liver after pulmonary or intramuscular administrations.

Wound healing is a complex process that can be severely impaired due to pathological situations such as diabetes mellitus. Diabetic foot ulcers are a common complication of this pathology and are characterized by an excessive inflammatory response. In this work, the effects of local treatment with recombinant human epidermal growth factor (rhEGF) were studied using a full-thickness wound healing model in streptozotocin-induced diabetic rats. Wound healing process was assessed with different concentrations of rhEGF (0.1, 0.5, 2.0 and 8.0 µg/mL), placebo and both diabetic and non-diabetic controls (n = 53). The macroscopic healing observed in treated diabetic rats was affected by rhEGF concentration. Histologically, we also observed an improvement in the epithelialization, granulation tissue formation and maturation in treated groups, finding again the best response at doses of 0.5 and 2.0 µg/mL. Afterwards, the tissue immune response over time was assessed in diabetic rats using the most effective concentrations of rhEGF (0.5 and 2.0 µg/mL), compared to controls. The presence of macrophages, CD4+ T lymphocytes and CD8+ T lymphocytes, in the reparative tissue was quantified, and cytokine expression was measured by quantitative real-time PCR. rhEGF treatment caused a reduction in the number of infiltrating macrophages in the healing tissue of diabetic, as well as diminished activation of these leukocytes. These findings show that local administration of rhEGF improves the healing process of excisional wounds and the quality of the neoformed tissue in a dose-dependent manner. Besides, this treatment reduces the local inflammation associated with diabetic healing, indicating immuno-modulatory properties.

This paper presents a procedure for the determination of seven nitrophenols (NPs) in water and soil samples using stir bar sorptive extraction (SBSE) coupled to gas chromatography with mass spectrometry (GC-MS) by means of a thermal desorption unit (TDU). Microwave assisted extraction (MAE) is proposed to release the NPs from the soil matrices into an aqueous phase, prior to their acetylation. The different variables affecting the preconcentration efficiency of SBSE, during both the adsorption and the thermal desorption steps, are studied. As regards the analytical characteristics of the method, the accuracy was measured through recovery studies, re covery percentages in all cases being in the 79–120% range, as well as by analyzing a certified reference ma terial. The precision was evaluated in terms of relative standard deviation, which provided values lower than 15% for both repeatability and reproducibility. The limits of detection were between 0.001 and 0.031 µg L−1 for water and 0.020–0.107 ng g−1 for soil samples. When environmental samples of different origins were analyzed, contents in the 0.01–1.0 µg L−1 and 0.7–40 ng g−1 ranges were obtained for waters and soils, respectively.

A sensitive method for the determination of five new generation pesticides (chlorantraniliprole, spirotetramat, spiromesifen, spirodiclofen and flubendiamide) in soil samples has been developed using liquid chromatography and tandem mass spectrometry (LC-MS 2 ) with a triple-quadrupole in the multiple reaction monitoring mode. The target analytes are released from the solid matrix by single-phase extraction in acetonitrile (SLE), this organic phase being used as dispersant solvent in the dispersive liquid–liquid microextraction (DLLME) step. The different parameters affecting the extraction efficiency in DLLME were carefully studied, being 1.5 mL of acetonitrile extract (disperser solvent), 125 μL carbon tetrachloride (extraction solvent) and 10 mL aqueous solution, the selected conditions. The enriched organic phase was evaporated, reconstituted with 50 μL acetonitrile and injected into a liquid chromatograph with a mobile phase composed of acetonitrile and 0.1% (v/v) formic acid under gradient elution and a C 8 stationary phase. Detection limits in the 0.0015–0.0090 ng g −1 range were obtained. Insecticide concentrations ranging from 0.03 to 197 ng g −1 , depending on the compound, were found in the soil samples analysed. The recovery values obtained by SLE-DLLME-LC-ESI-MS 2 for three spiked soils at three concentration levels varied between 87 and 114%, with RSDs of between 5.5 and 14%.

Phthalic acid esters (PEs) were preconcentrated from cleaning products, detergents and cosmetics using ultrasound assisted extraction (UAE) in the presence of acetonitrile, and then submitted to dispersive liquid-liquid microextraction (DLLME). For DLLME, 3mL of acetonitrile extract, 150μL carbon tetrachloride and 10mL aqueous solution were used. The enriched organic phase was evaporated, reconstituted with 25μL acetonitrile and injected into a liquid chromatograph with a mobile phase (acetonitrile:10mM ammonium acetate, pH 4) under gradient elution. Detection was carried out using both diode-array (DAD) and electrospray-ion trap-tandem mass spectrometry (ESI-IT-MS/MS) in the multiple reaction monitoring mode (MRM) of the positive fragment ions. Quantification was carried out using matrix-matched standards. Detection limits were in the range 0.04-0.45ngmL(-1) for the six PEs considered. The recoveries obtained were in the 84-124% range, with RSDs lower than 10%. Thirty three different cleaning products were analyzed. The most frequently found compound was diethyl phthalate.

© 2014 Elsevier B.V. All rights reserved. Lung impairment is the most life-threatening factor for cystic fibrosis patients. Indeed, Pseudomonas aeruginosa is the main pathogen in the pulmonary infection of these patients. In this work, we developed sodium colistimethate loaded lipid nanoparticles, namely, solid lipid nanoparticles (SLN) and nanostructured lipid carriers (NLC), as a strategy to enhance the antimicrobial therapy against P. aeruginosa in cystic fibrosis patients. The nanoparticles obtained displayed a 200-400 nm size, high drug entrapment (79-94%) and a sustained drug release profile. Moreover, both SLN and NLC presented antimicrobial activity against clinically isolated P. aeruginosa. The integrity of the nanoparticles was not affected by nebulization through a mesh vibrating nebulizer. Moreover, lipid nanoparticles appeared to be less toxic than free sodium colistimethate in cell culture. Finally, an in vivo distribution experiment showed that nanoparticles spread homogenously through the lung and there was no migration of lipid nanoparticles to other organs, such as liver, spleen or kidneys., This work was carried out under the TERFIQEC Project, Cromprehensive Research On Effective Therapies For The Treatment Of Cystic Fibrosis And Associated Diseases; IPT-2011-1402-900000 was funded by the Ministry of Economy and Competitiveness. Technical and human support provided by SGIker (UPV/EHU, MICINN, GV/EJ, ERDF and ESF) is gratefully acknowledged. M. Moreno-Sastre thanks the University of the Basque Country for the ZabaldUz fellowship grant. The authors gratefully acknowledge the support of University of the Basque Country UPV/EHU (UFI11/32), of University of Barcelona (UB) as to the CSIC and FISIB