First case report of multivisceral transplant from a deceased cardiac death donor.

The current shortage of pediatric multivisceral donors accounts for the long time and mortality on the waiting list of pediatric patients. The use of donors after cardiac death, especially after the outbreak of normothermic regional perfusion, has increased in recent years for all solid organs except the intestine, mainly because of its higher susceptibility to ischemia-reperfusion injury. We present the first literature case of multivisceral donors after cardiac death transplantation in a 13-month-old recipient from a 2.5-month-old donor. Once exitus was certified, an extracorporeal membrane oxygenation circuit was established, cannulating the aorta and infrarenal vena cava, while the supra-aortic branches were clamped. The abdominal organs completely recovered from ischemia through normothermic regional perfusion (extracorporeal membrane oxygenation initially and beating heart later). After perfusion with the preservation solution, the multivisceral graft was uneventfully implanted. Two months later, the patient was discharged without any complications. This case demonstrates the possibility of reducing the time spent on the waiting list for these patients.

Regulation of the Glycine Transporter GLYT1 by microRNAs.

The glycine transporter GLYT1 participates in inhibitory and excitatory neurotransmission by controlling the reuptake of this neuroactive substance from synapses. Over the past few years, microRNAs have emerged as potent negative regulators of gene expression. In this report, we investigate the possible regulation of GLYT1 by microRNAs. TargetScan software predicted the existence of multiple targets for microRNAs within the 3' UTR of the human GLYT1 (miR-7, miR-30, miR-96, miR-137 and miR-141), and as they are all conserved among mammalian orthologues, their effects on GLYT1 expression were determined experimentally. Dual reporter bioluminescent assays showed that only miR-96 and miR-137 down-regulated expression of the Renilla reporter fused to the 3' UTR of GLYT1. Mutations introduced into the target sequences blocked this inhibitory effect. Consistently, these two microRNAs downregulated the uptake of [3H]glycine into glial C6 cells, a cell line where GLYT1 is the main carrier for glycine. Moreover, the expression of endogenous GLYT1 in primary mixed cultures from rat spinal cord was decreased upon lentiviral expression of miR-96 and miR-137. Although the bulk of GLYT1 is glial, it is abundantly expressed in glycinergic neurons of the retina and in smaller amounts in glutamatergic neurons though the brain. Since miR-96 in the retina is strongly downregulated by light exposure, when rats were maintained in darkness for a few hours we observed a concomitant increase of GLYT1 expression, suggesting that at least miR-96 might be an important negative regulator of GLYT1 under physiological conditions.

Calcium-Dependent Regulation of the Neuronal Glycine Transporter GlyT2 by M2 Muscarinic Acetylcholine Receptors.

The neuronal glycine transporter GlyT2 modulates inhibitory glycinergic neurotransmission and plays a key role in regulating nociceptive signal progression. The cholinergic system acting through muscarinic acetylcholine receptors (mAChRs) also mediates important regulations of nociceptive transmission being the M2 subtype the most abundantly expressed in the spinal cord. Here we studied the effect of M2 mAChRs stimulation on GlyT2 function co-expressed in a heterologous system with negligible levels of muscarinic receptor activity. We found GlyT2 is down-regulated by carbachol in a calcium-dependent manner. Different components involved in cell calcium homeostasis were analysed to establish a role in the mechanism of GlyT2 inhibition. GlyT2 down-regulation by carbachol was increased by thapsigargin and reduced by internal store depletion, although calcium release from endoplasmic reticulum or mitochondria had a minor role on GlyT2 inhibition. Our results are consistent with a GlyT2 sensitivity to intracellular calcium mobilized by M2 mAChRs in the subcortical area of the plasma membrane. A crucial role of the plasma membrane sodium calcium exchanger NCX is proposed.

Metalliferous Mining Pollution and Its Impact on Terrestrial and Semi-terrestrial Vertebrates: A Review.

Metalliferous mining, a major source of metals and metalloids, has severe potential environmental impacts. However, the number of papers published in international peer-reviewed journals seems to be low regarding its effects in terrestrial wildlife. To the best of our knowledge, our review is the first on this topic. We used 186 studies published in scientific journals concerning metalliferous mining or mining spill pollution and their effects on terrestrial and semi-terrestrial vertebrates. We identified the working status of the mine complexes studied, the different biomarkers of exposure and effect used, and the studied taxa. Most studies (128) were developed in former mine sites and 46 in active mining areas. Additionally, although several mining accidents have occurred throughout the world, all papers about effects on terrestrial vertebrates from mining spillages were from Aznalcóllar (Spain). We also observed a lack of studies in some countries with a prominent mining industry. Despite >50% of the studies used some biomarker of effect, 42% of them only assessed exposure by measuring metal content in internal tissues or by non-invasive sampling, without considering the effect in their populations. Most studied species were birds and small mammals, with a negligible representation of reptiles and amphibians. The information gathered in this review could be helpful for future studies and protocols on the topic and it facilitates a database with valuable information on risk assessment of metalliferous mining pollution.

Overexpression of P2X3 and P2X7 Receptors and TRPV1 Channels in Adrenomedullary Chromaffin Cells in a Rat Model of Neuropathic Pain.

We have tested the hypothesis that neuropathic pain acting as a stressor drives functional plasticity in the sympathoadrenal system. The relation between neuropathic pain and adrenal medulla function was studied with behavioral, immunohistochemical and electrophysiological techniques in rats subjected to chronic constriction injury of the sciatic nerve. In slices of the adrenal gland from neuropathic animals, we have evidenced increased cholinergic innervation and spontaneous synaptic activity at the splanchnic nerve⁻chromaffin cell junction. Likewise, adrenomedullary chromaffin cells displayed enlarged acetylcholine-evoked currents with greater sensitivity to α-conotoxin RgIA, a selective blocker of α9 subunit-containing nicotinic acetylcholine receptors, as well as increased exocytosis triggered by voltage-activated Ca2+ entry. Altogether, these adaptations are expected to facilitate catecholamine output into the bloodstream. Last, but most intriguing, functional and immunohistochemical data indicate that P2X3 and P2X7 purinergic receptors and transient receptor potential vanilloid-1 (TRPV1) channels are overexpressed in chromaffin cells from neuropathic animals. These latter observations are reminiscent of molecular changes characteristic of peripheral sensitization of nociceptors following the lesion of a peripheral nerve, and suggest that similar phenomena can occur in other tissues, potentially contributing to behavioral manifestations of neuropathic pain.

A novel dominant hyperekplexia mutation Y705C alters trafficking and biochemical properties of the presynaptic glycine transporter GlyT2.

Hyperekplexia or startle disease is characterized by an exaggerated startle response, evoked by tactile or auditory stimuli, producing hypertonia and apnea episodes. Although rare, this orphan disorder can have serious consequences, including sudden infant death. Dominant and recessive mutations in the human glycine receptor (GlyR) α1 gene (GLRA1) are the major cause of this disorder. However, recessive mutations in the presynaptic Na(+)/Cl(-)-dependent glycine transporter GlyT2 gene (SLC6A5) are rapidly emerging as a second major cause of startle disease. In this study, systematic DNA sequencing of SLC6A5 revealed a new dominant GlyT2 mutation: pY705C (c.2114A→G) in transmembrane domain 11, in eight individuals from Spain and the United Kingdom. Curiously, individuals harboring this mutation show significant variation in clinical presentation. In addition to classical hyperekplexia symptoms, some individuals had abnormal respiration, facial dysmorphism, delayed motor development, or intellectual disability. We functionally characterized this mutation using molecular modeling, electrophysiology, [(3)H]glycine transport, cell surface expression, and cysteine labeling assays. We found that the introduced cysteine interacts with the cysteine pair Cys-311-Cys-320 in the second external loop of GlyT2. This interaction impairs transporter maturation through the secretory pathway, reduces surface expression, and inhibits transport function. Additionally, Y705C presents altered H(+) and Zn(2+) dependence of glycine transport that may affect the function of glycinergic neurotransmission in vivo.

An aspartate residue in the external vestibule of GLYT2 (glycine transporter 2) controls cation access and transport coupling.

Synaptic glycine levels are controlled by GLYTs (glycine transporters). GLYT1 is the main regulator of synaptic glycine concentrations and catalyses Na+-Cl--glycine co-transport with a 2:1:1 stoichiometry. In contrast, neuronal GLYT2 supplies glycine to the presynaptic terminal with a 3:1:1 stoichiometry. We subjected homology models of GLYT1 and GLYT2 to molecular dynamics simulations in the presence of Na+. Using molecular interaction potential maps and in silico mutagenesis, we identified a conserved region in the GLYT2 external vestibule likely to be involved in Na+ interactions. Replacement of Asp471 in this region reduced Na+ affinity and Na+ co-operativity of transport, an effect not produced in the homologous position (Asp295) in GLYT1. Unlike the GLYT1-Asp295 mutation, this Asp471 mutant increased sodium leakage and non-stoichiometric uncoupled ion movements through GLYT2, as determined by simultaneously measuring current and [3H]glycine accumulation. The homologous Asp471 and Asp295 positions exhibited distinct cation-sensitive external accessibility, and they were involved in Na+ and Li+-induced conformational changes. Although these two cations had opposite effects on GLYT1, they had comparable effects on accessibility in GLYT2, explaining the inhibitory and stimulatory responses to lithium exhibited by the two transporters. On the basis of these findings, we propose a role for Asp471 in controlling cation access to GLYT2 Na+ sites, ion coupling during transport and the subsequent conformational changes.

Job Insecurity and Company Behavior: Influence of Fear of Job Loss on Individual and Work Environment Factors.

The purpose of this research is to analyze to what extent job insecurity is related to different factors related with quality of work life. Specifically, it refers to the individual (work-family balance, job satisfaction, labor and professional development, motivation at work, and well-being at work) and work environment (conditions and environment and safety and health at work) dimensions of the construct. The sample group consisted of 842 workers (375 men and 467 women), aged between 18 and 68 years, from Bahía de Banderas, Mexico. Pearson correlation coefficients between the different variables were carried out, as well as MANOVA and ANOVA analyses and a linear regression analysis. The results showed that workers with low job insecurity obtained higher scores in work-family balance, job satisfaction, labor and professional development, motivation at work, well-being at work, conditions and environment, and safety and health at work, in relation to workers with moderate and high insecurity. The regression analysis confirmed that individual factors explain 24% and environmental factors 15% of job insecurity. This article makes an approximation to the phenomenon of job insecurity in the Mexican context, where the relationship of this variable with quality of work life is verified.

P2Y purinergic regulation of the glycine neurotransmitter transporters.

The sodium- and chloride-coupled glycine neurotransmitter transporters (GLYTs) control the availability of glycine at glycine-mediated synapses. The mainly glial GLYT1 is the key regulator of the glycine levels in glycinergic and glutamatergic pathways, whereas the neuronal GLYT2 is involved in the recycling of synaptic glycine from the inhibitory synaptic cleft. In this study, we report that stimulation of P2Y purinergic receptors with 2-methylthioadenosine 5'-diphosphate in rat brainstem/spinal cord primary neuronal cultures and adult rat synaptosomes leads to the inhibition of GLYT2 and the stimulation of GLYT1 by a paracrine regulation. These effects are mainly mediated by the ADP-preferring subtypes P2Y(1) and P2Y(13) because the effects are partially reversed by the specific antagonists N(6)-methyl-2'-deoxyadenosine-3',5'-bisphosphate and pyridoxal-5'-phosphate-6-azo(2-chloro-5-nitrophenyl)-2,4-disulfonate and are totally blocked by suramin. P2Y(12) receptor is additionally involved in GLYT1 stimulation. Using pharmacological approaches and siRNA-mediated protein knockdown methodology, we elucidate the molecular mechanisms of GLYT regulation. Modulation takes place through a signaling cascade involving phospholipase C activation, inositol 1,4,5-trisphosphate production, intracellular Ca(2+) mobilization, protein kinase C stimulation, nitric oxide formation, cyclic guanosine monophosphate production, and protein kinase G-I (PKG-I) activation. GLYT1 and GLYT2 are differentially sensitive to NO/cGMP/PKG-I both in brain-derived preparations and in heterologous systems expressing the recombinant transporters and P2Y(1) receptor. Sensitivity to 2-methylthioadenosine 5'-diphosphate by GLYT1 and GLYT2 was abolished by small interfering RNA (siRNA)-mediated knockdown of nitric-oxide synthase. Our data may help define the role of GLYTs in nociception and pain sensitization.

Impact of Bullying-Victimization and Gender over Psychological Distress, Suicidal Ideation, and Family Functioning of Mexican Adolescents.

Bullying victimization is strongly associated with increased psychological distress and suicide in adolescents and poor family functioning. Knowledge of gender differences influencing these factors will improve the prevention of mental problems and suicide in victimized adolescents. A total of 1685 Mexican secondary students, 12-17 years old (m = 13.65), of whom 54% were girls, responded to a standardized scale questionnaire to analyze such differences. Based on the statistical analysis, girls reported significantly lower family functioning and higher psychological distress and suicidal ideation than boys. The cluster analysis classified adolescents into high (5.78%), moderate (24.07%), and no-victimization (69.76%) groups. Boys predominated in the high (3.1%) and moderate-victimization (12.4%) clusters, and girls in the no-victimization group (39.51%). Multivariate statistical analyses found significant differences between the three groups, with the highest means of psychological distress and suicidal ideation and lowest family functioning in the high-victimization group. Only for suicidal ideation, there was an interaction between gender and the degree of victimization, with girls showing a higher increase of suicidal ideation than boys in the same cluster. Conclusions: Early detection and intervention in bullying-victimized adolescents, aiming to decrease psychological distress and suicidal ideation and strengthen family functioning, should consider contextual gender differences for effective prevention of mental health problems and suicide in adolescents.

Subcellular localization of the neuronal glycine transporter GLYT2 in brainstem.

The neuronal glycine transporter GLYT2 belongs to the neurotransmitter:sodium:symporter (NSS) family and removes glycine from the synaptic cleft, thereby aiding the termination of the glycinergic signal and achieving the reloading of the presynaptic terminal. The task fulfilled by this transporter is fine tuned by regulating both transport activity and intracellular trafficking. Different stimuli such as neuronal activity or protein kinase C (PKC) activation can control GLYT2 surface levels although the intracellular compartments where GLYT2 resides are largely unknown. Here, by biochemical and immunological techniques in combination with electron and confocal microscopy, we have investigated the subcellular distribution of GLYT2 in rat brainstem tissue, and characterized the vesicles that contain the transporter. GLYT2 is shown to be present in small and larger vesicles that contain the synaptic vesicle protein synaptophysin, the recycling endosome small GTPase Rab11, and in the larger vesicle population, the vesicular inhibitory amino acid transporter VIAAT. Rab5A, the GABA transporter GAT1, synaptotagmin2 and synaptobrevin2 (VAMP2) were not present. Coexpression of a Rab11 dominant negative mutant with recombinant GLYT2 impaired transporter trafficking and glycine transport. Dual immunogold labeling of brainstem synaptosomes showed a very close proximity of GLYT2 and Rab11. Therefore, the intracellular GLYT2 resides in a subset of endosomal membranes and may traffic around several compartments, mainly Rab11-positive endosomes.

Temporal trends of inorganic elements in a common kestrel (Falco tinnunculus) population from south west Spain.

Historical variations (1966-2016) in the exposure to inorganic elements in raptors of South West Spain have been little studied. Therefore, uncertainty exists concerning whether environmental or anthropogenic inputs, as well as dietary strategy shift, may cause changes in exposure patterns. To address this gap, essential and non-essential inorganic elements were measured in Common kestrel (Falco tinnunculus) feathers from museum specimens and free-living individuals spanning a 50-year period. Moreover, stable isotopes of δ13C and δ15N were also measured as proxies of Common kestrel feeding ecology over time. In general, all elements showed significant increasing trends overtime, suggesting changes of inputs from local sources. Moreover, δ15N signatures were correlated to the variations in Al, Cr, Cu, Fe, K, Li, Mg, Mn, Ni, P, Pb, Rb, Sr, Ti, V and Zn concentrations, suggesting trophic changes in this kestrel population as well as possible biomagnification processes. Finally, some values of Cr, Mn and Pb were above the threshold concentrations of potential toxicity.

Why Are Spanish Nurses Going to Work Sick? Questionnaire for the Measurement of Presenteeism in Nurses.

Presenteeism is defined as the presence of the worker at their workplace despite not being in optimal physical or mental conditions. Presenteeism is a phenomenon that has been poorly studied in the context of healthcare. Despite the many negative consequences associated with presenteeism, to date, no studies have investigated this issue in nurses in Spain. The objective was to develop and validate a questionnaire on presenteeism to be used by nursing staff in Spain. METHODS: A psychometric study for the development and validation of a questionnaire. The PRESENCA® questionnaire on presenteeism was created by a panel of experts, based on a survey comprised of 31 Likert-type items. RESULTS: In total, 355 nurses completed the questionnaire. The factorial analysis revealed the existence of 3 factors and confirmed appropriate levels of validity and reliability (alpha = 0.729). CONCLUSIONS: The PRESENCA® questionnaire is the first tool developed and validated in Spanish for the assessment of presenteeism in nursing. Our findings demonstrate that this scale has appropriate psychometric properties and its use may facilitate the detection of presenteeism among professionals. As a result, use of this questionnaire may contribute towards the improvement of clinical safety.

[Contribution of the occupational health to the COVID-19 pandemic control.] / Contribución de la salud laboral al control de la pandemia por covid-19 en España.

When the World Health Organization declared Covid-19 as a public health emergency of international concern, the Spanish Ministry of Health called the health, labor, social security authorities, Labor and Social Security Inspection, National Institute of Security and Occupational Health, employers, unions, occupational risk prevention services, mutual societies and scientific societies of occupational medicine and nursing, to collaborate in the control of the transmission of SARS-CoV-2 in companies. The Occupational Health Group of the Public Health Commission of the Interterritorial Council of the National Health System, developed the Procedure for the prevention of occupational risks in the face of exposure to SARS-CoV-2, which has been updated 15 times until the date. It contains the prevention measures to be implemented in the workplaces: organizational and collective protection, personal protection, especially vulnerable worker and risk level, study and management of cases and contacts that occurred in the company, collaboration in the management of temporary disability and, more recently, reincorporation and management of vaccinated workers. As a result of these cooperation and collaboration frameworks, a series of activities were deployed in the workplace, which are described in this article.

Cuando la Organización Mundial de la Salud declaró la Covid-19 como una emergencia de salud pública de importancia internacional, el Ministerio de Sanidad convocó a las autoridades sanitarias, laborales, de seguridad social, Inspección de Trabajo y Seguridad Social, Instituto Nacional de Seguridad y Salud en el Trabajo, empresarios, sindicatos, servicios de prevención de riesgos laborales, mutuas y sociedades científicas de la medicina y enfermería del trabajo, para colaborar en el control de la transmisión del SARS-CoV-2 en el ámbito de las empresas. La Ponencia de Salud Laboral de la Comisión de Salud Pública del Consejo Interterritorial del Sistema Nacional de Salud, elaboró el Procedimiento para los servicios de prevención de riesgos laborales frente a la exposición al SARS-CoV-2, que se ha actualizado 15 veces hasta la fecha. En él se recogen las medidas de prevención a implantar en los centros de trabajo: de carácter organizativo y de protección colectiva, de protección personal, de trabajador especialmente vulnerable y nivel de riesgo, de estudio y manejo de casos y contactos ocurridos en la empresa, de colaboración en la gestión de la incapacidad temporal y, más recientemente, de reincorporación y gestión de las y los trabajadores vacunados. Como resultado de esos marcos de cooperación y colaboración se desplegaron una serie de actividades en los lugares de trabajo que son descritas en este artículo.

Feathers and hair as tools for non-destructive pollution exposure assessment in a mining site of the Iberian Pyrite Belt.

Mining is responsible of releasing trace elements to the environment with potential negative effects on wildlife. Traditionally, wildlife exposure assessment has been developed by analyzing mainly environmental compartments or internal tissues. Nowadays, the use of non-destructive matrices such as hair or feathers has increased. Nevertheless, its use in free-living terrestrial mammals or in birds other than raptors or passerines is less frequent. The main objective of our study was to determine the potential for hair and feathers in a rabbit and bird species to be used as non-invasive proxy tissues for assessing internal metal concentrations at polluted sites from mining. We tested whether hair of European rabbit (Oryctolagus cuniculus) and feathers of red-legged partridge (Alectoris rufa) can be used as non-destructive biological monitoring tools of both essential (Cu, Zn) and non-essential (Pb, Cd, As) trace elements in a currently active copper mining site. We found significant different concentrations, particularly in non-essential elements, between reference area and mining site. Non-essential elements Pb and Cd showed higher correlations between tissues and hair/feathers, while few significant patterns were observed for essential elements such as Cu and Zn. Although feathers showed lower levels of correlation with internal tissues than hair, both could be useful as non-destructive biological monitoring tools. Further tissues, and more importantly, hair and feathers allowed discrimination between polluted and reference sites to indicate bioavailability and pollution status. In addition, hair and feathers can be used in monitoring pollution of an active mining site, being specially interesting for biomonitoring a certain period of time in the event of a particular episode of pollution, in addition to the chronic exposure. As occurred with hair in rabbits, feathers seem to be a good compartment to detect differences between a potential polluted area, such the surrounding of an active mine site, and a non-polluted area.

Job Insecurity, Family Functionality and Mental Health: A Comparative Study between Male and Female Hospitality Workers.

The aim of the present article is to compare the family functionality, mental health and job insecurity of employees of the hospitality industry in Puerto Vallarta and Bahía de Banderas, in México. This is a quantitative and cross-sectional study. The sample was selected by non-probabilistic sampling for convenience and comprised a total of 914 people, of whom 438 were women (47.92%) and 476 were men (52.08%). The women surveyed reported more somatic symptoms, anxiety, insomnia and social dysfunction compared to men, which allows the conclusion that their mental health is vulnerable; meanwhile, men showed better perception of family functionality, a positive factor that reveals the family as a potential support factor that reduces stress, anxiety and improves men's mental health. Another result reveals that the gender structure permeates the hotel sector, inequalities in the type of contract and income are corroborated, and the existence of a sexual division of labor to the detriment of women is confirmed, as they are mostly employed in low-skilled jobs that reproduce domestic tasks, particularly those related to cleaning and food service tasks.

Bullying Victimization among Mexican Adolescents: Psychosocial Differences from an Ecological Approach.

This transversal study over a random representative sample of 1687 Mexican students attending public and private secondary schools (54% girls, 12-17 years old, M = 13.65. DT = 1.14) aimed to analyze psychosocial differences between victims and non-victims of bullying from the bioecological model. It included individual variables (ontosystem), familiar, community, and scholar factors (microsystem), and gender (macrosystem) to perform a multivariate discriminant analysis and a logistic regression analysis. The discriminant analysis found that psychological distress, offensive communication with mother and father, and a positive attitude toward social norms transgression characterized the high victimization cluster. For the non-victims, the discriminant variables were community implication, positive attitude toward institutional authority, and open communication with the mother. These variables allowed for correctly predicting membership in 76% of the cases. Logistic regression analysis found that psychological distress, offensive communication with the father, and being a boy increased the probability of high victimization, while a positive attitude toward authority, open communication with the mother, and being a girl decrease this probability. These results highlight the importance of open and offensive communication between adolescents and their parents on psychological distress, attitude toward authority, community implication, and bullying victimization.

P2X receptors up-regulate the cell-surface expression of the neuronal glycine transporter GlyT2.

Glycinergic inhibitory neurons of the spinal dorsal horn exert critical control over the conduction of nociceptive signals to higher brain areas. The neuronal glycine transporter 2 (GlyT2) is involved in the recycling of synaptic glycine from the inhibitory synaptic cleft and its activity modulates intra and extracellular glycine concentrations. In this report we show that the stimulation of P2X purinergic receptors with ßγ-methylene adenosine 5'-triphosphate induces the up-regulation of GlyT2 transport activity by increasing total and plasma membrane expression and reducing transporter ubiquitination. We identified the receptor subtypes involved by combining pharmacological approaches, siRNA-mediated protein knockdown, and dorsal root ganglion cell enrichment in brainstem and spinal cord primary cultures. Up-regulation of GlyT2 required the combined stimulation of homomeric P2X3 and P2X2 receptors or heteromeric P2X2/3 receptors. We measured the spontaneous glycinergic currents, glycine release and GlyT2 uptake concurrently in response to P2X receptor agonists, and showed that the impact of P2X3 receptor activation on glycinergic neurotransmission involves the modulation of GlyT2 expression or activity. The recognized pro-nociceptive action of P2X3 receptors suggests that the fine-tuning of GlyT2 activity may have consequences in nociceptive signal conduction.

The role of density-dependence regulation in the misleading effect of the Aznalcollar mining spill on the booted eagle fecundity.

The aim of this study was to assess the exposure and potential effects of the toxic elements released by the Aznalcollar spillage on the fecundity of the booted eagle (Hieraetus pennatus) population of Doñana National Park. We measured As, Cd, Cu, Pb, Zn, Sb and Tl levels in the blood of 24 individuals after the accident and also used data of reproductive performance of the species from 1976 to 2000. While As, Sb and Tl were below their detection limits and only 3 samples were above detection limit for Cd (0.02-0.04µg/dl), mean levels of Cu, Zn and Pb (µg/dl) were found to be 38.736, 532.937 and 0.719, respectively, in fledglings; and 66.112, 652.571 and 1.825, respectively, in adults. These levels were higher in adults, but they did not differ significantly between age classes and were at safe levels. In addition, a positive relationship between relative fecundity and the combination of Cu, Zn and Pb was observed. Following the mining spill, the mean population fecundity was lower (0.536 fledglings per pair in the population) than before (1.093). However, the decrease in fecundity over time was also related to the increase of breeding pairs, suggesting a density-dependent regulation and nullifying the mining spillage effect. Thus, although raptors have proved to be highly valuable biomonitors of environmental pollution, we must be especially cautious and go beyond the apparent effects that sometimes we quickly believe to detect, leading us to reach mistaken conclusions that could affect our management plans.