Advances in the study of cylindrospermopsin: detection of potentially cylindrospermopsin-producing cyanobacteria, and prospect for the initial steps of the toxin synthesis

Tesis doctoral inédita, leída en Universidad Autónoma de Madrid, Facultad de Ciencias, Departamento de Biología. Fecha de lectura: 19-12-201

Characterization of Oxidative Lipidomics and Autophagy Induction in Chlamydomonas reinhardtii Under Abiotic Stress

Autophagy constitutes an essential process triggered upon by oxidative stress that enables cells to recycle damaged biomolecules and organelles, which is eventually traced by immunodetection with anti-ATG8. In parallel with autophagy induction, carbon metabolism in Chlamydomonas reinhardtii under abiotic stress is diverged toward lipid biosynthesis and lipid droplet accumulation, which can be analyzed by a simple thin-layer chromatography and in vivo staining with the fluorescent probe BODIPY 493/503. We show the responses in Chlamydomonas cells exposed to mercury or cadmium (0–50 μM doses), as examples of oxidative stress-mediated changes in autophagy and lipid metabolism, monitored with the procedures described in this repor

Fluorescent in vivo imaging of reactive oxygen species and redox potential in plants

Reactive oxygen species (ROS) are by-products of aerobic metabolism, and excessive production can result in oxidative stress and cell damage. In addition, ROS function as cellular messengers, working as redox regulators in a multitude of biological processes. Understanding ROS signalling and stress responses requires methods for precise imaging and quantification to monitor local, subcellular and global ROS dynamics with high selectivity, sensitivity and spatiotemporal resolution. In this review, we summarize the present knowledge for in vivo plant ROS imaging and detection, using both chemical probes and fluorescent protein-based biosensors. Certain characteristics of plant tissues, for example high background autofluorescence in photosynthetic organs and the multitude of endogenous antioxidants, can interfere with ROS and redox potential detection, making imaging extra challenging. Novel methods and techniques to measure in vivo plant ROS and redox changes with better selectivity, accuracy, and spatiotemporal resolution are therefore desirable to fully acknowledge the remarkably complex plant ROS signalling networksThis work was funded by a grant from the Spanish Ministry of Economy and Competitiveness ( AGL2014–53771-R ). Alfonso Blázquez-Castro acknowledges funding under the Marie Skłodowska-Curie Action COFUND 2015 (EU project 713366 – InterTalentum

Attenuation of mercury phytotoxicity with a high nutritional level of nitrate in alfalfa plants grown hydroponically

Mercury (Hg) is one of the most dangerous pollutant heavy metals to the environment, which causes several toxic effects in plants upon accumulation, such as induction of oxidative stress. Nitrate (NO3 – ) is the prevalent form to incorporate nitrogen (N) in higher plants, through its reduction to nitrite (NO2 – ) by the enzyme nitrate reductase (NR). We studied the physiological alterations caused by Hg (0, 6 and 30 µM) in alfalfa plants grown at two different levels of NO3 – : low, (2 mM; LN), and high (12 mM; HN) for one week using a semi-hydroponic culture system. Several parameters of oxidative stress such as lipid peroxidation, chlorophyll content, biothiol concen tration, and ascorbate peroxidase (APX) and glutathione reductase (GR) activities showed that HN plants were less affected by Hg. Nitrate reductase activity and NO3 – concentration were also altered under Hg stress, with lower impact in plants nourished with high NO3 – . Our results highlight the importance of the NO3 – nutritional status to improve tolerance to toxic metals like H

Synchrotron Radiation-Fourier transformed infrared microspectroscopy (μSR-FTIR) reveals multiple metabolism alterations in microalgae induced by cadmium and mercury

Toxic metals such as cadmium (Cd) and mercury (Hg) represent a threat to photosynthetic organisms of polluted aquatic ecosystems, and knowledge about mechanisms of toxicity is essential for appropriate assessment of environmental risks. We used Synchrotron Radiation-Fourier Transformed Infrared microspectroscopy (μSR-FTIR) to characterise major changes of biomolecules caused by Cd and Hg in the model green microalga Chlamydomonas reinhardtii. μSR-FTIR showed several metabolic alterations in different biochemical groups such as carbohydrates, proteins, and lipids in a time-dose dependent manner, with the strongest changes occurring at concentrations above 10 μM Cd and 15 μM Hg after short-term (24 h) treatments. This occurred in a context where metals triggered intracellular oxidative stress and chloroplast damage, along with autophagy induction by overexpressing AUTOPHAGY-RELATED PROTEIN 8 (ATG8). Thin layer chromatography analysis confirmed that toxic metals promoted remarkable changes in lipid profile, with higher degree of esterified fatty acid unsaturation as detected by gas chromatography coupled with mass spectrometry. Under Cd stress, there was specifically higher unsaturation of free fatty acids, while Hg led to stronger unsaturation in monogalactosyldiacylglycerol. μSR-FTIR spectroscopy proved as a valuable tool to identify biochemical alterations in microalgae, information that could be exploited to optimise approaches for metal decontaminationWork supported by the Spanish State Research Agency (AEI) (Spain) (projects AGL2014–53771-R and AGL2017–87591-R). The FTIR experiments were performed at MIRAS beamline at ALBA Synchrotron with the collaboration of ALBA staff and supported by travel grants for experiments No 2016091860 and 201702211

Fibers spreading worldwide: Microplastics and other anthropogenic litter in an Arctic freshwater lake

We investigated the presence of microplastics and other anthropogenic litter in the sediments adhered to rocks of an Arctic freshwater lake at Ny-Ålesund (Svalbard Archipelago, 78°N; 11°E). Most of the sampled microparticles were fibers (>90%). The identification of polymer types and additives was performed by combining three spectroscopic techniques, namely Raman Microscopy, Fourier-Transform Infrared microspectroscopy (μFTIR) and Synchrotron Radiation μFTIR (SR-FTIR). SR-FTIR confirmed the presence of poly(ethylene terephthalate) fibers, while RAMAN spectroscopy provided evidence of fibers containing industrial additives. Our results estimated an average concentration of 400 microparticles/m2 of rocks identified as anthropogenic litter, which included an estimation of 90 microplastics/m2 identified as polyester fibers; the rest are mostly natural fibers with evidence of anthropogenic origin. Taken together, the results proved the occurrence of anthropogenic pollutants in remote polar areas. Their probable origin is the long range atmospheric transpor

Mitos, creencias y temores en la cardiopatía isquémica

Los pacientes que sufren cardiopatía isquémica conviven con frecuencia con mitos y falsas creencias al respecto de su nueva situación; que pueden traer consecuencias que se pueden manifestar en forma de una incorrecta prevención de procesos posteriores y el desarrollo de prácticas y modos de vida nocivos para el individuo. Este estudio de diseño cualitativo realizado sobre pacientes con enfermedad coronaria aguda ingresados en nuestra unidad de cuidados críticos pretende evaluar el grado de conocimiento de la población que sufre esta enfermedad coronaria con respecto a la gravedad, calidad de vida y cuidados preventivos y paliativos que manejaban estos usuarios, revisando temas como la estima, las creencias, la sexualidad, los roles familiares y laborales, etc. Las conclusiones extraídas del análisis de los datos ofrecidos por los pacientes entrevistados muestran que existe un claro desconocimiento con respecto a su situación, sus posibilidades y sus perspectivas, que los introducen en marcos erróneos que traen como consecuencia el aumento de sus miedos y preocupaciones, siendo éste el mayor causante de estrés y preocupaciones, con consecuencias negativas para su evolución y afrontamiento a la nueva situació

Neuromyelitis optica spectrum disorders. Comparison according to the phenotype and serostatus

Objective: To (1) determine the value of the recently proposed criteria of neuromyelitis optica (NMO) spectrum disorder (NMOSD) that unify patients with NMO and those with limited forms (NMO/LF) with aquaporin-4 immunoglobulin G (AQP4-IgG) antibodies; and (2) investigate the clinical significance of the serologic status in patients with NMO. Methods: This was a retrospective, multicenter study of 181 patients fulfilling the 2006 NMO criteria (n = 127) or NMO/LF criteria with AQP4-IgG (n = 54). AQP4-IgG and myelin oligodendrocyte glycoprotein immunoglobulin G (MOG-IgG) antibodies were tested using cell-based assays. Results: Patients were mainly white (86%) and female (ratio 6.5:1) with median age at onset 39 years (range 10-77). Compared to patients with NMO and AQP4-IgG (n = 94), those with NMO/LF presentedmore often with longitudinally extensive transverse myelitis (LETM) (p<0.001), and had lower relapse rates (p = 0.015), but similar disability outcomes. Nonwhite ethnicity and optic neuritis presentation doubled the risk for developing NMO compared with white race (p = 0.008) or LETM presentation (p = 0.008). Nonwhite race (hazard ratio [HR] 4.3, 95% confidence interval [CI] 1.4-13.6) and older age at onset were associated with worse outcome (for every 10-year increase, HR 1.7, 95% CI 1.3-2.2). Patients with NMO and MOG-IgG (n = 9) had lower female: male ratio (0.8:1) and better disability outcome than AQP4-IgG-seropositive or double-seronegative patients (p<0.001). Conclusions: In patients with AQP4-IgG, the similar outcomes regardless of the clinical phenotype support the unified term NMOSD; nonwhite ethnicity and older age at onset are associated with worse outcome. Double-seronegative and AQP4-IgG-seropositive NMO have a similar clinical outcome. The better prognosis of patients with MOG-IgG and NMO suggests that phenotypic and serologic classification is useful

Risk profiles and one-year outcomes of patients with newly diagnosed atrial fibrillation in India: Insights from the GARFIELD-AF Registry.

BACKGROUND: The Global Anticoagulant Registry in the FIELD-Atrial Fibrillation (GARFIELD-AF) is an ongoing prospective noninterventional registry, which is providing important information on the baseline characteristics, treatment patterns, and 1-year outcomes in patients with newly diagnosed non-valvular atrial fibrillation (NVAF). This report describes data from Indian patients recruited in this registry. METHODS AND RESULTS: A total of 52,014 patients with newly diagnosed AF were enrolled globally; of these, 1388 patients were recruited from 26 sites within India (2012-2016). In India, the mean age was 65.8 years at diagnosis of NVAF. Hypertension was the most prevalent risk factor for AF, present in 68.5% of patients from India and in 76.3% of patients globally (P < 0.001). Diabetes and coronary artery disease (CAD) were prevalent in 36.2% and 28.1% of patients as compared with global prevalence of 22.2% and 21.6%, respectively (P < 0.001 for both). Antiplatelet therapy was the most common antithrombotic treatment in India. With increasing stroke risk, however, patients were more likely to receive oral anticoagulant therapy [mainly vitamin K antagonist (VKA)], but average international normalized ratio (INR) was lower among Indian patients [median INR value 1.6 (interquartile range {IQR}: 1.3-2.3) versus 2.3 (IQR 1.8-2.8) (P < 0.001)]. Compared with other countries, patients from India had markedly higher rates of all-cause mortality [7.68 per 100 person-years (95% confidence interval 6.32-9.35) vs 4.34 (4.16-4.53), P < 0.0001], while rates of stroke/systemic embolism and major bleeding were lower after 1 year of follow-up. CONCLUSION: Compared to previously published registries from India, the GARFIELD-AF registry describes clinical profiles and outcomes in Indian patients with AF of a different etiology. The registry data show that compared to the rest of the world, Indian AF patients are younger in age and have more diabetes and CAD. Patients with a higher stroke risk are more likely to receive anticoagulation therapy with VKA but are underdosed compared with the global average in the GARFIELD-AF. CLINICAL TRIAL REGISTRATION-URL: http://www.clinicaltrials.gov. Unique identifier: NCT01090362

Mortality from gastrointestinal congenital anomalies at 264 hospitals in 74 low-income, middle-income, and high-income countries: a multicentre, international, prospective cohort study

Summary Background Congenital anomalies are the fifth leading cause of mortality in children younger than 5 years globally. Many gastrointestinal congenital anomalies are fatal without timely access to neonatal surgical care, but few studies have been done on these conditions in low-income and middle-income countries (LMICs). We compared outcomes of the seven most common gastrointestinal congenital anomalies in low-income, middle-income, and high-income countries globally, and identified factors associated with mortality. Methods We did a multicentre, international prospective cohort study of patients younger than 16 years, presenting to hospital for the first time with oesophageal atresia, congenital diaphragmatic hernia, intestinal atresia, gastroschisis, exomphalos, anorectal malformation, and Hirschsprung’s disease. Recruitment was of consecutive patients for a minimum of 1 month between October, 2018, and April, 2019. We collected data on patient demographics, clinical status, interventions, and outcomes using the REDCap platform. Patients were followed up for 30 days after primary intervention, or 30 days after admission if they did not receive an intervention. The primary outcome was all-cause, in-hospital mortality for all conditions combined and each condition individually, stratified by country income status. We did a complete case analysis. Findings We included 3849 patients with 3975 study conditions (560 with oesophageal atresia, 448 with congenital diaphragmatic hernia, 681 with intestinal atresia, 453 with gastroschisis, 325 with exomphalos, 991 with anorectal malformation, and 517 with Hirschsprung’s disease) from 264 hospitals (89 in high-income countries, 166 in middleincome countries, and nine in low-income countries) in 74 countries. Of the 3849 patients, 2231 (58·0%) were male. Median gestational age at birth was 38 weeks (IQR 36–39) and median bodyweight at presentation was 2·8 kg (2·3–3·3). Mortality among all patients was 37 (39·8%) of 93 in low-income countries, 583 (20·4%) of 2860 in middle-income countries, and 50 (5·6%) of 896 in high-income countries (p<0·0001 between all country income groups). Gastroschisis had the greatest difference in mortality between country income strata (nine [90·0%] of ten in lowincome countries, 97 [31·9%] of 304 in middle-income countries, and two [1·4%] of 139 in high-income countries; p≤0·0001 between all country income groups). Factors significantly associated with higher mortality for all patients combined included country income status (low-income vs high-income countries, risk ratio 2·78 [95% CI 1·88–4·11], p<0·0001; middle-income vs high-income countries, 2·11 [1·59–2·79], p<0·0001), sepsis at presentation (1·20 [1·04–1·40], p=0·016), higher American Society of Anesthesiologists (ASA) score at primary intervention (ASA 4–5 vs ASA 1–2, 1·82 [1·40–2·35], p<0·0001; ASA 3 vs ASA 1–2, 1·58, [1·30–1·92], p<0·0001]), surgical safety checklist not used (1·39 [1·02–1·90], p=0·035), and ventilation or parenteral nutrition unavailable when needed (ventilation 1·96, [1·41–2·71], p=0·0001; parenteral nutrition 1·35, [1·05–1·74], p=0·018). Administration of parenteral nutrition (0·61, [0·47–0·79], p=0·0002) and use of a peripherally inserted central catheter (0·65 [0·50–0·86], p=0·0024) or percutaneous central line (0·69 [0·48–1·00], p=0·049) were associated with lower mortality. Interpretation Unacceptable differences in mortality exist for gastrointestinal congenital anomalies between lowincome, middle-income, and high-income countries. Improving access to quality neonatal surgical care in LMICs will be vital to achieve Sustainable Development Goal 3.2 of ending preventable deaths in neonates and children younger than 5 years by 2030