Estudio de la utilidad clínica del perfil de estrés oxidativo en pacientes con déficit de alfa-1 antitripsina

Introducción: Investigaciones recientes en modelos animales sugieren la participación del estrés oxidativo y el daño de oxidativo en la patogénesis del déficit de alfa 1 antitripsina (DAAT). Sin embargo, no hay datos disponibles sobre el estado de estrés oxidativo y la actividad de los enzimas antioxidantes en estos pacientes. Se sabe que el estrés oxidativo está implicado en la patogénesis de enfermedades en otros organismos (aterosclesosis, enfermedades neurodegenerativas, diabetes mellitus, enfermedades reumáticas, etc). También que el tabaco es un importante oxidante, disminuyendo la afinidad de la alfa-1 antitripsina, principal antiproteasa del organismo, por la elastasa del neutrófilo, pudiendo favorecer la enfermedad pulmonar (enfisema) en pacientes con DAAT al existir un desequilibrio proteasa-antiproteasa; aunque hay un reducido grupo de enfermos no fumadores que también presentan esta enfermedad, por lo que se plantea el estudio con el fin de analizar el perfil de estrés oxidativo, los mecanismos enzimáticos de defensa antioxidante y parámetros de daño oxidativo en una población pediátrica sin manifestaciones clínicas de la enfermedad y sin factores distorsionadores, como puede ser el hábito tabáquico, con DAAT. Métodos: Se determina el perfil de estrés oxidativo, la actividad de las principales enzimas antioxidantes (catalasa, glutatión peroxidasa, glutatión reductasa y superóxido dismutasa) así como el daño oxidativo a nivel del ADN (determinación de 8 hidroxideoxiguanosina -8-OHdG-), lípidos (malondialdéhido -MDA-) y proteinas (proteínas carboniladas) en el suero de cincuenta y cuatro niños diagnosticados de déficit de alfa-1 antitripsina (según los criterios diagnósticos de las guias internacionales) y treinta y cinco niños incluidos como grupo control (sin déficit de alfa-1 antitripsina con fenotipo MM). Los pacientes con DAAT fueron agrupados en base al riesgo de enfermedad pulmonar y/o hepática correlacionado con el fenotipo en: bajo riesgo (MS y SS), riesgo intermedio (MZ y SZ) y alto riesgo (ZZ) Resultados: El estrés oxidativo está aumentado en el suero de niños con riesgo intermedio (MZ y SZ) y alto (ZZ) para desarrollar enfisema y/o enfermedad hepática relacionado con el déficit de alfa-1 antitripsina. Cuando se compara con el grupo control, los grupos de riesgo intermedio y alto presentan diferencias estadísticamente significativas caracterizadas por: disminución de los niveles de glutatión total y glutatión reducido, así como disminución de la actividad de la enzima catalasa y aumento de la actividad de la enzima glutatión peroxidasa, todo ello conduce a un acúmulo de peróxido de hidrógeno celular, lo cual explicaría el aumento significativo de los niveles de marcadores biológicos de daño oxidativo (8-OHdG, MDA y proteinas carboniladas) observado en estos pacientes. No se encuentran diferencias significativas al comprar el grupo control (MM) con el grupo de bajo riesgo (MS y SS). Además, al comparar a los pacientes con déficit de alfa-1 antitripsina entre sí, se observa una gradación en los parámetros de estrés oxidativo, actividad enzimática y marcadores de daño oxidativo, objetivando que la expresión del alelo Z produce un estado de estrés oxidativo mayor en pacientes homocigotos (ZZ) que en heterocigotos (MZ; SZ). Conclusiones: El aumento de estrés oxidativo, junto con la disminución de las defensas antioxidantes, y el aumento de parámetros de daño oxidativo podrían estar implicados en la fisiopatología del DAAT en tempranas etapas; fundamentalmente en los pacientes que presentan alelo Z en su fenotipo

Adaptation to bronchial asthma in pediatrics: anxiety and psychological well-being

Marco teórico: el asma bronquial es la enfermedad crónica más prevalente en la infancia y adolescencia. Su presencia es un factor de riesgo para desarrollar un trastorno psicológico, siendo la ansiedad la sintomatología más común en estos pacientes. Por otro lado, el bienestar psicológico es un factor de protección para un adecuado ajuste a la enfermedad. Nuestro objetivo es analizar los niveles de ansiedad y de bienestar psicológico en adolescentes con asma bronquial. Métodos: la muestra incluye 73 adolescentes asmáticos de 9 a 18 años. Se analizaron las variables Ansiedad (HADS) y Bienestar Psicológico (BIEPS-J) mediante análisis descriptivos, pruebas t para muestras independientes y correlaciones de Pearson. Resultados: un tercio de la muestra presenta niveles elevados de ansiedad (diagnóstico probable= 9,60%; caso probable= 26%). Las medias obtenidas en la escala BIEPS-J fueron altas, encontrando diferencias significativas en el factor “aceptación de sí mismo” (t=2,42, p=,02, d=,57), siendo mayor en los hombres. No se han encontrado relaciones significativas entre las variables. Conclusiones: resulta necesario esclarecer los factores que influyen en la existencia de niveles elevados de bienestar psicológico en dichos pacientes y tomar medidas sanitarias de prevención multidisciplinares con el fin de detectar tempranamente la presencia de ansiedad y tomar actuaciones psicológicas pertinentesTheoretical Framework:bronchial asthma is the most prevalent chronic disease in childhood and adolescence. Its presence is a risk factor to develop a psychological disorder, with anxiety being the most common symptomatology in these patients. On the other hand, psychological well-being is a protective factor for an adequate adjustment to the disease. Our objective is to analyze levels of anxiety and psychological well-being in adolescents with bronchial asthma.Methods:sample includes 73 asthmatic adolescents from 9 to 18 years old. The variables Anxiety (HADS) and Psychological Wellbeing (BIEPS-J) were analyzed by descriptive analysis, independent samples t-test and Pearson correlations. Results:one third of the sample presented high levels of anxiety (probable diagnosis = 9.60%, probable case = 26%). The means obtained in the BIEPS-J scale were high, finding significant differences in the factor "acceptance of oneself" (t = 2.42, p = .02, d = .57), being higher in men. No significant relationships were found between the variables.Conclusions:it is necessary to clarify the factors that influence the existence of high levels of psychological well-being in these patients and to take sanitary multidisciplinary prevention measures to detect early the presence of anxiety and take relevant psychological action

Protective factors in the adjustment to cystic fibrosis in paediatrics

Marco teórico: la fibrosis quística es la enfermedad rara más frecuente. Su diagnóstico tiene un impacto sobre el bienestar delos pacientes. El objetivo de nuestro estudio es analizar los factores de protección en el ajuste a la enfermedad.Métodos:seevaluaron25 pacientes pediátricos (60% chicas), de edades entre 9-18 años (M=11,52, DT= 2,86). Se administraron dos cuestionarios: Fortalezas y dificultades y la Escala de Bienestar Psicológico. Se realizaron análisis descriptivos y correlaciones de Pearson entre las variables. Resultados: los datos obtenidos evidenciaron que un 44% de los pacientes presentaba un bajo bienestar psicológico, sin embargo, la mayoría mostraron una alta conducta prosocial. Existían relaciones positivas entre el bienestar psicológico y la prosocialidad. Conclusiones: conocer los aspectos protectores para un adecuado ajuste a la enfermedad, resulta relevante para desarrollar programas que potencien estos aspectos y favorezcan el bienestar del paciente.Theoretical Framework:cystic fibrosis is the most common rare disease. Their diagnosis has an impact on the well-being of patients. The objective of our study is to analyze the protective factors in the adjustment to the diseaseMethods:25 paediatric patients (60% female), aged 9-18 years (M=11.52, DT=2.86) were evaluated. Two questionnaires were administered: Strengths and difficulties and the Psychological Well-being Scale. Descriptive analyses and Pearson correlations between variables were performed.Results:the data obtained showed that 44% of the patients presented a low psychological well-being, however, the majority showed a high prosocial behaviour. There were positive relationships between psychological well-being and prosociality.Conclusions:knowing the protective aspects for an adequate adjustment to the disease is relevant to develop programs that seek to enhance these aspects and promote the patient's well-bein

Comparison of Physical Activity and Sedentary Behaviour between Schoolchildren with Cystic Fibrosis and Healthy Controls: A Gender Analysis

The purpose of this study was to examine differences in sports participation and the levels of physical activity (PA) and sedentary behaviour (SB) between schoolchildren with cystic fibrosis (CF) and a healthy control group (CG) taking into account the gender variable. PA and SB were measured with an accelerometer for 7 consecutive days in 44 children (24 girls; 11.0 (3.2) years) with CF and 45 age-, sex-, and socioeconomic status-matched controls (24 girls; 11.1 (3.0) years). CF patients and CG did not differ in moderate-to-vigorous PA (54 (31) vs. 59 (27) min/day respectively) or in SB (558 (106) vs. 553 (92) min/day respectively). There were no differences in meeting the PA guidelines between both groups (CF: 36.4% vs. CG: 42.4%). Gender analysis revealed that boys were more active and met more PA guidelines than girls regardless of the group (CF or CG), girls with CF being the least active group (only 16.7% met PA guidelines). A possible compensatory effect was found between SB and PA only in the CF sample, as for each minute/day spent in SB the odds of meeting PA guidelines decreased by 34%. These findings suggest that promoting a reduction in SB is as important as promoting PA in the CF population, especially in girls. Health caregivers, coaches, teachers, or parents could offer appealing supervised and unsupervised physical activities, foster the adoption of active lifestyles, or incorporate PA into daily routines to improve the health of CF schoolchildren

Pulmonary function testing in children's interstitial lung disease

The use of pulmonary function tests (PFTs) has been widely described in airway diseases like asthma and cystic fibrosis, but for children's interstitial lung disease (chILD), which encompasses a broad spectrum of pathologies, the usefulness of PFTs is still undetermined, despite widespread use in adult interstitial lung disease. A literature review was initiated by the COST/Enter chILD working group aiming to describe published studies, to identify gaps in knowledge and to propose future research goals in regard to spirometry, whole-body plethysmography, infant and pre-school PFTs, measurement of diffusing capacity, multiple breath washout and cardiopulmonary exercise tests in chILD. The search revealed a limited number of papers published in the past three decades, of which the majority were descriptive and did not report pulmonary function as the main outcome.PFTs may be useful in different stages of management of children with suspected or confirmed chILD, but the chILD spectrum is diverse and includes a heterogeneous patient group in all ages. Research studies in well-defined patient cohorts are needed to establish which PFT and outcomes are most relevant for diagnosis, evaluation of disease severity and course, and monitoring individual conditions both for improvement in clinical care and as end-points in future randomised controlled trials

The disease-specific clinical trial network for primary ciliary dyskinesia: PCD-CTN

Primary ciliary dyskinesia (PCD) is a rare genetic disorder characterised by impaired mucociliary clearance leading to irreversible lung damage. In contrast to other rare lung diseases like cystic fibrosis (CF), there are only few clinical trials and limited evidence-based treatments. Management is mainly based on expert opinions and treatment is challenging due to a wide range of clinical manifestations and disease severity. To improve clinical and translational research and facilitate development of new treatments, the clinical trial network for PCD (PCD-CTN) was founded in 2020 under the framework of the European Reference Network (ERN)-LUNG PCD Core. Applications from European PCD sites interested in participating in the PCD-CTN were requested. Inclusion criteria consisted of patient numbers, membership of ERN-LUNG PCD Core, use of associated standards of care, experience in PCD and/or CF clinical research, resources to run clinical trials, good clinical practice (GCP) certifications and institutional support. So far, applications from 22 trial sites in 18 European countries have been approved, including >1400 adult and >1600 paediatric individuals with PCD. The PCD-CTN is headed by a coordinating centre and consists of a steering and executive committee, a data safety monitoring board and committees for protocol review, training and standardisation. A strong association with patient organisations and industrial companies are further cornerstones. All participating trial sites agreed on a code of conduct. As CTNs from other diseases have demonstrated successfully, this newly formed PCD-CTN operates to establish evidence-based treatments for this orphan disease and to bring new personalised treatment approaches to patients

Redox Imbalance in Nasal Epithelial Cells of Primary Ciliary Dyskinesia Patients

Background: Primary Ciliary Dyskinesia (PCD) represents a rare condition marked by an abnormal mobility pattern of cilia and flagella, resulting in impaired mucociliary clearance. This deficiency leads to recurrent infections and persistent inflammation of the airways. While previous studies have indicated heightened oxidative stress levels in the exhaled breath condensate of pediatric PCD patients, the assessment of oxidative stress within the affected respiratory tissue remains unexplored. Aims: To assess the oxidative status of human nasal epithelial cells (NECs) in PCD patients. Methods: Thirty-five PCD patients and thirty-five healthy control subjects were prospectively included in the study. Levels of reactive oxygen species (ROS), reactive nitrogen species (RNS), glutathione (GSH), intracellular Ca2+, plasma membrane potential, and oxidative damage in lipids and proteins were measured. In addition, apoptosis and mitochondrial function were analyzed by flow cytometry in NECs. Results: NECs from PCD patients showed reduced levels of apoptosis (p = 0.004), superoxide anion (O2−, p = 0.018), peroxynitrite (ONOO−, p = 0.007), nitric oxide (NO, p = 0.007), mitochondrial hydrogen peroxide (mtH2O2, p 2−, p = 0.0004) and increased mitochondrial mass (p = 0.009) compared to those from healthy individuals. No significant differences were observed in oxidized proteins (p = 0.137) and the oxidized/reduced lipid ratio (p = 0.7973). The oxidative profile of NEC cells in PCD patients, according to their ciliary motility, recurrent otitis, recurrent pneumonia, atelectasis, bronchiectasis, and situs inversus, showed no statistically significant differences in the parameters studied. Conversely, patients with chronic rhinosinusitis exhibited lower levels of ONOO− than PCD patients without this condition, with no significant differences related to other symptoms. Conclusions: Our findings strongly suggest the presence of a redox imbalance, specifically leaning toward a reductive state, in PCD patients

Hypoxia Enhances Oxidative Stress in Neutrophils from ZZ Alpha-1 Antitrypsin Deficiency Patients

Alpha-1 antitrypsin deficiency (AATD) is a neutrophilic inflammatory disorder that may result in local hypoxia, reactive oxygen and nitrogen species (ROS/RNS) production, and increased damage in adjacent tissues. This study aims to determine the impact of hypoxia on neutrophil oxidative stress profile in AATD patients. Neutrophils were isolated from AATD patients and control volunteers and exposed to hypoxia (1% O2 for 4 h), ROS/RNS, mitochondrial parameters, and non-enzymatic antioxidant defenses measured by flow cytometry. The expression of enzymatic antioxidant defenses was determined by qRT-PCR. Our results indicate that ZZ-AATD neutrophils produce higher amounts of hydrogen peroxide, peroxynitrite, and nitric oxide and decreased levels of the antioxidant enzymes catalase, superoxide dismutase, and glutathione reductase. Likewise, our results show a decrease in mitochondrial membrane potential, indicating that this organelle could be involved in the production of the reactive species observed. No decrease in glutathione and thiol levels were observed. The accumulation of substances with high oxidative capacity would explain the greater oxidative damage observed in proteins and lipids. In conclusion, our results indicate that, compared to MM control individuals, ZZ-AATD neutrophils show increased ROS/RNS production under hypoxic conditions opening a new rationale for using antioxidant therapies to treat the disease

Immunofluorescence Analysis as a Diagnostic Tool in a Spanish Cohort of Patients with Suspected Primary Ciliary Dyskinesia

Primary ciliary dyskinesia (PCD) is an autosomal recessive rare disease caused by an alteration of ciliary structure. Immunofluorescence, consisting in the detection of the presence and distribution of cilia proteins in human respiratory cells by fluorescence, has been recently proposed as a technique to improve understanding of disease-causing genes and diagnosis rate in PCD. The objective of this study is to determine the accuracy of a panel of four fluorescently labeled antibodies (DNAH5, DNALI1, GAS8 and RSPH4A or RSPH9) as a PCD diagnostic tool in the absence of transmission electron microscopy analysis. The panel was tested in nasal brushing samples of 74 patients with clinical suspicion of PCD. Sixty-eight (91.9%) patients were evaluable for all tested antibodies. Thirty-three cases (44.6%) presented an absence or mislocation of protein in the ciliary axoneme (15 absent and 3 proximal distribution of DNAH5 in the ciliary axoneme, 3 absent DNAH5 and DNALI1, 7 absent DNALI1 and cytoplasmatic localization of GAS8, 1 absent GAS8, 3 absent RSPH9 and 1 absent RSPH4A). Fifteen patients had confirmed or highly likely PCD but normal immunofluorescence results (68.8% sensitivity and 100% specificity). In conclusion, immunofluorescence analysis is a quick, available, low-cost and reliable diagnostic test for PCD, althouh it cannot be used as a standalone tes

Implementation of a Gene Panel for Genetic Diagnosis of Primary Ciliary Dyskinesia

Introduction: Primary ciliary dyskinesia (PCD) is characterized by an alteration in the ciliary structure causing difficulty in the clearance of respiratory secretions. Diagnosis is complex and based on a combination of techniques. The objective of this study was to design a gene panel including all known causative genes, and to corroborate their diagnostic utility in a cohort of Spanish patients. Methods: This was a multicenter cross-sectional study of patients with a high suspicion of PCD, according to European Respiratory Society criteria, designed around a gene panel for massive sequencing using SeqCap EZ capture technology that included 44 genes associated with PCD. Results: We included 79 patients, 53 of whom had a diagnosis of confirmed or highly probable PCD. The sensitivity of the gene panel was 81.1%, with a specificity of 100%. Candidate variants were found in some of the genes of the panel in 43 patients with PCD, 51.2% (22/43) of whom were homozygotes and 48.8% (21/43) compound heterozygotes. The most common causative genes were DNAH5 and CCDC39. We found 52 different variants, 36 of which were not previously described in the literature. Conclusions: The design and implementation of a tailored gene panel produces a high yield in the genetic diagnosis of PCD. This panel provides a better understanding of the causative factors involved in these patients and lays down the groundwork for future therapeutic approaches