A walk in the PARC: developing and implementing 21st century chemical risk assessment in Europe.

Current approaches for the assessment of environmental and human health risks due to exposure to chemical substances have served their purpose reasonably well. Nevertheless, the systems in place for different uses of chemicals are faced with various challenges, ranging from a growing number of chemicals to changes in the types of chemicals and materials produced. This has triggered global awareness of the need for a paradigm shift, which in turn has led to the publication of new concepts for chemical risk assessment and explorations of how to translate these concepts into pragmatic approaches. As a result, next-generation risk assessment (NGRA) is generally seen as the way forward. However, incorporating new scientific insights and innovative approaches into hazard and exposure assessments in such a way that regulatory needs are adequately met has appeared to be challenging. The European Partnership for the Assessment of Risks from Chemicals (PARC) has been designed to address various challenges associated with innovating chemical risk assessment. Its overall goal is to consolidate and strengthen the European research and innovation capacity for chemical risk assessment to protect human health and the environment. With around 200 participating organisations from all over Europe, including three European agencies, and a total budget of over 400 million euro, PARC is one of the largest projects of its kind. It has a duration of seven years and is coordinated by ANSES, the French Agency for Food, Environmental and Occupational Health & Safety.

[Reproducibilidad clínica de reacciones de hipersensibilidad a fármacos en pruebas de exposición controlada].

Objective: To assess the reproducibility of symptoms in drug challenge tests. Methods: The study included patients with positive cutaneous or challenge test throughout 2019. For each patient, clinical suspicion according to Karch-Lasagna algorithm was registered. Primary outcome was the reproducibility of symptoms in the provocation tests using a paired analysis of data with McNemar test. Results: Eighty-nine patients were included, 16 of them presented more than one positive test. Thirty were skin tests positive and 75 reacted to provocation tests. Eighty nine percent of patients who reacted in challenge test were probably or possibly reactors according to Karch-Lasagna scale. Symptoms of initial reaction did not differ from those triggered in challenge tests. Conclusions: Karch-Lasagna scale is useful in predicting the response to drug provocation tests. In most of the positive studies, results were suggested by clinical history and no differences were found between symptoms triggered in challenge test and that referred to in the previous reaction.

Objetivo: Evaluar la reproducibilidad de los síntomas en pruebas de exposición con fármacos. Métodos: Estudio retrospectivo, efectuado en pacientes con prueba cutánea o exposición positiva, atendidos en 2019. De cada paciente se registró la sospecha clínica según el algoritmo de Karch-Lasagna. El resultado principal fue la reproducibilidad de síntomas en las pruebas de exposición, mediante el análisis de datos emparejado con prueba de McNemar. Resultados: Se incluyeron 89 pacientes, y de estos 16 reportaron varias pruebas positivas. Se obtuvieron 30 pruebas cutáneas y 75 de exposición positivas. En el 89% de las pruebas de exposición positivas, las reacciones iniciales se clasificaron en probables o posibles, según la escala de Karch-Lasagna. Los síntomas reportados en la reacción inicial no difirieron de los de las pruebas de exposición. Conclusiones: La escala de Karch-Lasagna es un método útil para predecir la respuesta en las pruebas de exposición con fármacos. En la mayor parte de las pruebas positivas, los resultados fueron sugeridos por la historia clínica, sin diferencias entre la manifestación de síntomas en la prueba de exposición versus los referidos en la reacción inicial.

Engineering platforms for directed evolution of Laccase from Pycnoporus cinnabarinus.

While the Pycnoporus cinnabarinus laccase (PcL) is one of the most promising high-redox-potential enzymes for environmental biocatalysis, its practical use has to date remained limited due to the lack of directed evolution platforms with which to improve its features. Here, we describe the construction of a PcL fusion gene and the optimization of conditions to induce its functional expression in Saccharomyces cerevisiae, facilitating its directed evolution and semirational engineering. The native PcL signal peptide was replaced by the α-factor preproleader, and this construct was subjected to six rounds of evolution coupled to a multiscreening assay based on the oxidation of natural and synthetic redox mediators at more neutral pHs. The laccase total activity was enhanced 8,000-fold: the evolved α-factor preproleader improved secretion levels 40-fold, and several mutations in mature laccase provided a 13.7-fold increase in k(cat). While the pH activity profile was shifted to more neutral values, the thermostability and the broad substrate specificity of PcL were retained. Evolved variants were highly secreted by Aspergillus niger (∼23 mg/liter), which addresses the potential use of this combined-expression system for protein engineering. The mapping of mutations onto the PcL crystal structure shed new light on the oxidation of phenolic and nonphenolic substrates. Furthermore, some mutations arising in the evolved preproleader highlighted its potential for heterologous expression of fungal laccases in yeast (S. cerevisiae).

Basophil activation test as alternative method to CAP-inhibition in patients with double sensitization to vespid venoms.

Most patients with Hymenoptera venom allergy (HVA) to vespid venoms present double sensitization by specific IgE (sIgE)-mediated cross-reactivity. Thus, it is mandatory could discriminate between a true double and primary sensitization to implement an accurate venom-specific immunotherapy (VIT). To date, CAP-inhibition is the reference method in the diagnosis of cross-reactivity in double sensitized patients to vespid venoms, being the results obtained with the component resolved diagnostics (CRD) conflicting. For this, we have studied in a cohort of double sensitized patients to Vespula vulgaris (VV) and Polistes dominulus (PD) venoms (n = 40) the diagnostic accuracy of CRD using the CAP-inhibition as reference method, as well as to investigate whether basophil activation test (BAT) is an alternative method for inconclusive results obtained by CAP-inhibition. CAP-inhibition showed a sensitivity of 59.46 % in view of the indeterminate results; most patients had true double sensitization (54.5 %), followed by single sensitization to PD (27.27 %) and VV (18.18 %) venoms. CRD based on rVes v 5/rPol d 5 (or vice versa) ratio as well as whole extracts I3/I77 (or vice versa) ratio (specific IgE-I3 to VV/specific IgE-I77 to PD) showed a low diagnostic accuracy (AUC = 0.504, p = 0.974; AUC = 0.35, p = 0.235; respectively). BAT was determined in parallel with CAP-inhibition in 12 patients, presented higher sensitivity than CAP-inhibition (p = 0.021) and a positive agreement of 71.43 %. Likewise it was able to identify 100% of inconclusive results, showing a specificity of 83.3 %. Therefore, CRD is not a suitable method to distinguish monosensitization and BAT appears to be an appropriate method resolving indeterminate results from the gold standard method.

Epigenetic control of the EWS­FLI1 promoter in Ewing's sarcoma.

Ewing sarcoma (ES) is a primary bone marrow tumor that very rarely develops in extra­osseous tissues, such as lung. The hallmark of ES tumors is a translocation between chromosomes 11 and 22, resulting in a fusion protein, commonly referred to as EWS­FLI1. The epigenetic profile (histone acetylation and methylation enrichment of the promoter region) that may regulate the expression of the aberrant transcription factor EWS­FLI1, remains poorly studied and understood. Knowledge of epigenetic patterns associated with covalent histone modification and expression of enzymes associated with this process, can contribute to the understanding of the molecular basis of the disease, as well as to the identification of possible molecular targets involved in expression of the EWS­FLI1 gene, so that therapeutic strategies may be improved in the future. In the present study, the transcriptional activation and repression of the EWS­FLI1 fusion gene in ES was accompanied by selective deposition of histone markers on its promoter. The EWS­FLI1 fusion gene was evaluated in two patients with ES using conventional cytogenetic, fluorescence in situ hybridization and nested PCR assays, which revealed that the aberrant expression of the EWS­FLI1 gene is accompanied by enrichment of H3K4Me3, H3K9ac and H3K27ac at the promoter region.

[Postanoxic encaphalopathy after and episode of resuscitated sudden death. Minimizing sequelae]. / Encefalopatía postanóxica tras episodio de muerte súbita reanimada. Cómo minimizar los daños.

Sudden death in young people while performing intense physical activity has a very low prevalence but a significant burden in terms of loss of years of life in society and a strong social impact. We present the case of a 19-year-old man who had a cardiac arrest while playing a football match, with prolonged resuscitation, and multiple subsequent complications (acute renal failure, coagulopathy, digestive bleeding, ischaemic colitis, and need for implantable cardioverter-defibrillator placement and hemicolectomy). The onset of intensive early rehabilitation in a specialised centre minimised the sequels, improving the Rankin score from 4 to 2 and Barthel index from 0 to 95 points, allowing the patient to lead an almost autonomous life.

Ex Vivo and In Vivo Biocompatibility Assessment (Blood and Tissue) of Three-Dimensional Bacterial Nanocellulose Biomaterials for Soft Tissue Implants.

Bacterial nanocellulose (BNC) is a promising biomedical material. However, the haemocompatibility (haemolysis and thrombogenicity) and acute and sub-chronic immune responses to three-dimensional (3D) BNC biomaterials have not been evaluated. Accordingly, this manuscript focused on the effect of 3D microporosity on BNC haemocompatibility and a comparison with 2D BNC architecture, followed by the evaluation of the immune response to 3D BNC. Blood ex vivo studies indicated that compared with other 2D and 3D BNC architectures, never-dried 2D BNC presented antihemolytic and antithrombogenic effects. Nevertheless, in vivo studies indicated that 3D BNC did not interfere with wound haemostasis and elicited a mild acute inflammatory response, not a foreign body or chronic inflammatory response. Moreover, compared with the polyethylene controls, the implant design with micropores ca. 60 µm in diameter showed a high level of collagen, neovascularization and low fibrosis. Cell/tissue infiltration increased to 91% after 12 weeks and was characterized by fibroblastic, capillary and extracellular matrix infiltration. Accordingly, 3D BNC biomaterials can be considered a potential implantable biomaterial for soft tissue augmentation or replacement.

Mercury levels in blood, urine and hair in a nation-wide sample of Spanish adults.

Mercury (Hg) is among the top 10 environmental chemicals of major public health concern (WHO). The Minamata Convention on Mercury (United Nations Environment Program, 2017), commits signing countries to control anthropogenic mercury emissions and reduce human exposure. Human biomonitoring (HBM) programs, are the most straight-forward approaches to get information on the actual exposure levels in the population and assess over time. We report here the results of a HBM study in a nationwide cross-section of Spanish adults (18-65y) as baseline values obtained before the Minamata Convention entered into force. Subsequent follow-ups will show if the Convention has been successful. The study includes 1880 blood samples, 1704 urine samples and 577 hair samples from all Spanish regions collected and analysed under a strictly quality controlled and quality assured protocol. The EU-DEMOCOPHES project demonstrated that fish and seafood are the major sources of mercury exposure and that the Spanish as well as the Portuguese populations have higher levels than other European countries. The data from the present study confirms this pattern at national level and that inhabitants in coastal regions have higher values than from inland regions. The geometric mean (GM) for blood is 6.35 µg Hg/l, in urine is 1.11 µg Hg/l and for hair is 1.91 µg Hg/g. In an international comparison these values are not exceptional. Spanish concentrations fall into the group of Easter Mediterranean populations. Although information on gender, age, occupational sector, geographical area, sampling period and frequency of fish consumption is reported in the tables, the purpose of this paper has not been to analyse the determinants of exposure in detail but to provide baseline data for future assessments and for regional authorities.

Effect of finasteride on human testicular steroidogenesis.

We studied the testicular function and some androgen-mediated events in 22 males (16-30 years of age) with male pattern baldness that was treated with finasteride (10 mg once daily) for 2 years. Patients were evaluated every 3 months. Prostatic volume was determined in six subjects by endorectal ultrasound scans. Serum gonadotropin, prostate-specific antigen (PSA), and sex hormone levels were determined basally and periodically during the treatment period. Fourteen subjects underwent gonadal stimulation with human chorionic gonadotropin (hCG), and the gonadotropin response to gonadotropin releasing hormone (GnRH) was determined in eight subjects, prior to and after 2 years of therapy. Finasteride treatment resulted in an improvement in the male pattern baldness and prostatic shrinkage that was associated with an increase in serum testosterone levels (17.2 +/- 2.5 vs. 26.3 +/- 1.7 nmol/L) and a decrease in dihydrotestosterone (DHT) levels (1.45 +/- 0.41 vs. 0.38 +/- 0.10 nmol/L), causing a marked increase in that testosterone/DHT ratio. A significant increase in the serum levels of androstenedione (3.67 +/- 0.49 vs. 7.05 +/- 0.70 nmol/L) and estradiol (132 +/- 44 vs. 187 +/- 26 pmol/L) was also noted, whereas androstanediol glucoronide (33.3 +/- 6.4 vs. 10.7 +/- 4.5 pmol) and PSA (1.6 +/- 0.6 vs. 0.4 +/- 0.1 ng/ml) were significantly decreased. No changes in basal or stimulated levels of gonadotropin were observed. There was a significant increase in the testosterone response to hCG during finasteride therapy (delta: 16.7 vs. 35.5 nmol/L) that could be explained, at least in part, by the reduction of testosterone metabolism resulting from the blockage induced by finasteride. The decrease in the androstenedione to testosterone and estrone to estradiol ratios observed after hCG treatment, however, strongly suggests increased activity of the 17-ketosteroid reductase enzyme and an improvement of the testicular capacity for testosterone production.

Differential characteristics of heart transplantation in patients older than 60 years.

BACKGROUND: The effect of advanced age on the results of heart transplantation (HTx) is still controversial. The few articles addressing this issue have not been conclusive, due to either short follow-up periods or small numbers of patients. METHODS: We present a retrospective study of 560 HTx which were divided into group A, including patients of 60 or less years at HTx (n=465, 83%), and group B, of 95 recipients older than 60 years. A subgroup of the latter, named B1, includes 24 patients older than 65. More than 100 recipient, donor and surgical procedure variables were analyzed for their impact on actuarial survival and incidence of common causes of posttransplant morbidity and mortality during a follow-up period longer than 10 years. RESULTS: Group B showed a lower number of acute rejection episodes than group A, (1.53+/-1.87 versus 1.96+/-1.81, P<.04). Both groups showed a similar incidence of infection episodes, malignancies or graft vasculopathy, but older patients experienced fewer viral infections than younger ones (9% in group A versus 18% in group B, P<.05). Log-rank test showed a trend to shorter survival in group B (P=.08), a disadvantage that reached significance (P=.01) among patients older than 65 years. CONCLUSIONS: Patients who were older than 60 at HTx displayed a lower incidence of acute rejection episodes and viral infections, but a trend toward shorter long-term survival. This disadvantage in prognosis was statistically significant among recipients older than 65 years.

Electrochemical characterization of an asymmetric nanofiltration membrane with NaCl and KCl solutions: influence of membrane asymmetry on transport parameters.

Electrochemical characterization of a nanofiltration asymmetric membrane was carried out by measuring membrane potential, salt diffusion, and electrical parameters (membrane electrical resistance and capacitance) with the membrane in contact with NaCl and KCl solutions at different concentrations (10(-3)< or =c(M)< or =5 x 10(-2)). From these experiments characteristic parameters such as the effective concentration of charge in the membrane, ionic transport numbers, and salt and ionic permeabilities across the membrane were determined. Membrane electrical resistance and capacitance were obtained from impedance spectroscopy (IS) measurements by using equivalent circuits as models. This technique allows the determination of the electrical contribution associated with each sublayer; then, assuming that the dense sublayer behaves as a plane capacitor, its thickness can be estimated from the capacitance value. The influence of membrane asymmetry on transport parameters have been studied by carrying out measurements for the two opposite external conditions. Results show that membrane asymmetry strongly affects membrane potential, which is attributed to the Donnan exclusion when the solutions in contact with the dense layer have concentrations lower than the membrane fixed charge (X(ef) approximately -0.004 M), but for the reversal experimental condition (high concentration in contact with the membrane dense sublayer) the membrane potential is practically similar to the solution diffusion potential. The comparison of results obtained for both electrolytes agrees with the higher conductivity of KCl solutions. On the other hand, the influence of diffusion layers at the membrane/solution interfaces in salt permeation was also studied by measuring salt diffusion at a given NaCl concentration gradient but at five different solutions stirring rates.

A comparison of electrochemical and electrokinetic parameters determined for cellophane membranes in contact with NaCl and NaNO3 solutions.

Electrochemical and electrokinetic characterizations of cellophane membrane samples have been carried out by measuring membrane potential, salt diffusion, and tangential streaming potential, which allow the determination of different characteristic membrane parameters. Experiments were made with the membrane samples in contact with NaCl and NaNO(3) solutions at different concentrations and under different external conditions (concentration gradients), in order to obtain differences in transport and membrane characteristic parameters, depending on the electrolyte considered. Salt permeability across the membrane, which was obtained from diffusion measurements, is about twice as high for NaCl solutions as for NaNO(3) solutions, which is attributed to the different sizes of the electrolytes. Membrane potential measurements keeping the concentration ratio constant (C(1)/C(2)=2) were used to determine both the effective fixed charge concentration in the membrane, X(f), and the average value of transport numbers, t(i); taking into account these values, concentration dependence of membrane potential under a different external condition (C(1)=cte=0.01 M, 5 x 10(-3)< or =C(M)< or =5 x 10(-2)) was predicted. Results show that cellophane membrane behaves as a weak cation-exchange membrane and its permselectivity to cations is practically independent of the electrolyte considered. From electrokinetic results, assuming a Langmuir-type adsorption of anions on the cellophane surface, the number of accessible sites per surface unit was obtained, which is higher for Cl(-) than for NO(3)(-), in agreement with the small radii of chlorine ions; however, no significant differences in the specific adsorption free energy were found (DeltaG(Nacl)=-22.0 x 10(3) J/mol) and (DeltaG(NaNO(3))=-23.2 x 10(3) J/mol).

[Resistance to antibiotics in clinical isolates of the genus Enterococcus]. / Resistencia a antibióticos en aislamientos clínicos del género Enterococcus.

The purpose of our study was to determine the prevalence of resistance in 114 clinical strains of enterococci. Identification was made using PASCO dehydrated panels and by the conventional method of Facklam and found the following: 92.1% E. faecalis, 7% E. faecium and 0.9% E. avium. The antibiotic susceptibility was determined by the PASCO system, and the minimum inhibitory concentrations of vancomycin, teicoplanin, gentamicin and streptomycin were determined by the agar dilution method. All the strains were susceptible to vancomycin and teicoplanin. We detected resistance to ampicillin in 4.4% of isolates, to penicillin in 6%, to tetracycline in 41.2%, to rifampicin in 12.3%, to chloramphenicol in 23.7%, to erythromycin in 66.7%, to ciprofloxacin in 53.5%, to gentamicin to a high level in 43.9%, and to streptomycin to a high level in 57%. E. faecium was associated with high-level resistance to gentamicin. E. faecium was more resistant than E. faecalis to quinolones, rifampicin, penicillin and ampicillin, and showed similar susceptibility to tetracycline and chloramphenicol. However, E. faecalis was more resistant than E. faecium to aminoglycosides, mainly to gentamicin.

Cytologic atypia. Clinical significance and follow-up recommendations.

OBJECTIVE: To assess the evolution of cytologic atypia on cervical cytologic smears as an attempt to better understand the natural history of these lesions and to establish a follow-up protocol. STUDY DESIGN: The study group consisted of 76 patients with a diagnosis of cytologic atypia on cervical cytologic smears performed at the gynecology Outpatient Clinic, Hospital del Mar, between December 1989 and June 1993. Patients with inflammatory atypia, a reparative process or findings consistent with human papillomavirus infection had been excluded. Follow-up smears were reviewed. Compliance with follow-up recommendations was also evaluated. RESULTS: Thirty-nine patients returned to the clinic for follow-up after a diagnosis of atypia (51.3%). Progression from atypia to dysplasia was found in 38.5% of patients with at least a follow-up smear. Changes consistent with intraepithelial neoplasia were found in 17 patients: low grade dysplasia was found in 10 patients (mild dysplasia in 8 and atypia in 2) and high grade dysplasia in 7 (moderate in 4 and severe in 3). This progression was most commonly found at the first follow-up visit and within the first year. CONCLUSION: Patients with cytologic atypia were at higher risk of developing dysplasia than those with reparative atypia. Our results support the theory of progression from atypia to dysplasia in a considerable percentage of patients. Follow-up compliance in our patient population was low. One should emphasize the importance of this diagnosis and encourage close cytologic follow-up, especially within the first year after the diagnosis of atypia.

[Chylothorax secondary to blunt thoracic trauma. Report of 6 cases]. / Quilotórax secundario a traumatismo torácico cerrado. A propósito de 6 casos.

Chylothorax is a rare condition that even more rarely arises as a result of closed thoracic trauma. We report a series of 6 patients who suffered chylothorax after closed trauma, who were diagnosed early and treated conservatively. Either total parenteral feeding or adjusted enteral feeding, depending with circumstances, was started early such that complications from chylothorax were few. Surgery was required in only one case, and outcomes were satisfactory in all patients, none of whom died.

[Traumatic bilateral chylothorax (author's transl)]. / Quilotórax bilateral traumático.

A case is presented of a bilateral chylothorax caused by rupture of the thoracic duct, secondary to a closed traumatism of the thorax in a polytraumatized patient, and produced by a mechanism of hyperextension of the dorsal segment of the vertebral column. Considerations are made with reference to the anatomy of the thoracic duct, the rupture mechanism, diagnosis, complications and treatment. The chylothorax could not be distinguished clinically and by simple radiology from the traumatic hemothorax. The preliminary diagnosis will be made according to the macroscopic appearance of the drained fluid, and the definite diagnosis by the analytical and histopathologic characteristics of the same fluid. The lymphographic examination reveals the exact point of the lesion. The immediate complications are related to the respiratory insufficiency due to the occupation of the pleural space, and under a long period of time undernutrition provoked by the loss of fats and proteins. The treatment of choice is based upon the early drainage of the chylothorax and on the absolute suppression of the oral intake, establishing a complete parenteral feeding during at least 15 days. Afterwards oral diet can be initiated with a gradual increase in its quantity and always without fat content, while at the same time the parenteral nutrition is reduced. Following this protocol a great number of ruptures of the thoracic duct can be corrected, achieving a spontaneous closure of the duct. In the cases in which this not occur, at the end of 15 days it would be advisable to perform a surgical operation according to the Lampson's ligature technique.

[Myocardial infarction due to buckshot lodged in the right coronary artery (author's transl)]. / Infarto de miocardio por perdigón alojado en arteria coronaria derecha.

A rare case of myocardial infarction produced by a buckshot lodged in the right coronary artery is presented. It resulted from a shotgun blast aimed at the right side of the chest. The patient was admitted to an Intensive Care Unit in a state of shock 1 hour after the accident. The diagnosis was suspected from the first moment because of ECG disturbances and it was later confirmed by coronary angiography. Hemodynamic studies were carried out simultaneously. The patient also presented hemothorax requiring pleural drainage. X-rays revealed a lung contusion with an intraparenchymatous hematoma the whole length of the projectile pathway. During the clinical course a slight hemopericardium was demonstrated by echocardiography; it was originally suspected on the basis of the X-rays. Drainage was not necessary. The patient left the Intensive Care Unit 7 days after entry and was discharged from the Hospital 5 days later. His clinical and hemodynamic condition was good and ECG indicate cicatrization of the myocardial infarction of the diaphragmatic face. The diagnosis and treatment of open or closed traumas of the thorax with similar cardiac involvement in emergency conditions are discussed. The authors prefer not to operate immediately in these cases.