Cell intercalation driven by SMAD3 underlies secondary neural tube formation.

Body axis elongation is a hallmark of the vertebrate embryo, involving the architectural remodeling of the tail bud. Although it is clear how neuromesodermal progenitors (NMPs) contribute to embryo elongation, the dynamic events that lead to de novo lumen formation and that culminate in the formation of a 3-dimensional, neural tube from NMPs, are poorly understood. Here, we used in vivo imaging of the chicken embryo to show that cell intercalation downstream of TGF-ß/SMAD3 signaling is required for secondary neural tube formation. Our analysis describes the events in embryo elongation including lineage restriction, the epithelial-to-mesenchymal transition of NMPs, and the initiation of lumen formation. We show that the resolution of a single, centrally positioned lumen, which occurs through the intercalation of central cells, requires SMAD3/Yes-associated protein (YAP) activity. We anticipate that these findings will be relevant to understand caudal, skin-covered neural tube defects, among the most frequent birth defects detected in humans.

In Vivo Analysis of the Mesenchymal-to-Epithelial Transition During Chick Secondary Neurulation.

The neural tube in amniotic embryos forms as a result of two consecutive events along the anteroposterior axis, referred to as primary and secondary neurulation (PN and SN). While PN involves the invagination of a sheet of epithelial cells, SN shapes the caudal neural tube through the mesenchymal-to-epithelial transition (MET) of neuromesodermal progenitors, followed by cavitation of the medullary cord. The technical difficulties in studying SN mainly involve the challenge of labeling and manipulating SN cells in vivo. Here we describe a new method to follow MET during SN in the chick embryo, combining early in ovo chick electroporation with in vivo time-lapse imaging. This procedure allows the cells undergoing SN to be manipulated in order to investigate the MET process, permitting their cell dynamics to be followed in vivo.

Multimerization of Zika Virus-NS5 Causes Ciliopathy and Forces Premature Neurogenesis.

Zika virus (ZikV) is a flavivirus that infects neural tissues, causing congenital microcephaly. ZikV has evolved multiple mechanisms to restrict proliferation and enhance cell death, although the underlying cellular events involved remain unclear. Here we show that the ZikV-NS5 protein interacts with host proteins at the base of the primary cilia in neural progenitor cells, causing an atypical non-genetic ciliopathy and premature neuron delamination. Furthermore, in human microcephalic fetal brain tissue, ZikV-NS5 persists at the base of the motile cilia in ependymal cells, which also exhibit a severe ciliopathy. Although the enzymatic activity of ZikV-NS5 appears to be dispensable, the amino acids Y25, K28, and K29 that are involved in NS5 oligomerization are essential for localization and interaction with components of the cilium base, promoting ciliopathy and premature neurogenesis. These findings lay the foundation for therapies that target ZikV-NS5 multimerization and prevent the developmental malformations associated with congenital Zika syndrome.

A centrosomal view of CNS growth.

Embryonic development of the central nervous system (CNS) requires the proliferation of neural progenitor cells to be tightly regulated, allowing the formation of an organ with the right size and shape. This includes regulation of both the spatial distribution of mitosis and the mode of cell division. The centrosome, which is the main microtubule-organizing centre of animal cells, contributes to both of these processes. Here, we discuss the impact that centrosome-mediated control of cell division has on the shape of the overall growing CNS. We also review the intrinsic properties of the centrosome, both in terms of its molecular composition and its signalling capabilities, and discuss the fascinating notion that intrinsic centrosomal asymmetries in dividing neural progenitor cells are instructive for neurogenesis. Finally, we discuss the genetic links between centrosome dysfunction during development and the aetiology of microcephaly.

Shh-mediated centrosomal recruitment of PKA promotes symmetric proliferative neuroepithelial cell division.

Tight control of the balance between self-expanding symmetric and self-renewing asymmetric neural progenitor divisions is crucial to regulate the number of cells in the developing central nervous system. We recently demonstrated that Sonic hedgehog (Shh) signalling is required for the expansion of motor neuron progenitors by maintaining symmetric divisions. Here we show that activation of Shh/Gli signalling in dividing neuroepithelial cells controls the symmetric recruitment of PKA to the centrosomes that nucleate the mitotic spindle, maintaining symmetric proliferative divisions. Notably, Shh signalling upregulates the expression of pericentrin, which is required to dock PKA to the centrosomes, which in turn exerts a positive feedback onto Shh signalling. Thus, by controlling centrosomal protein assembly, we propose that Shh signalling overcomes the intrinsic asymmetry at the centrosome during neuroepithelial cell division, thereby promoting self-expanding symmetric divisions and the expansion of the progenitor pool.

Changes in management of chronic obstructive pulmonary disease (COPD) in primary care: EMMEPOC study.

BACKGROUND: Changes in management of COPD in primary care were studied following a training exercise among Spanish general practitioners (GPs). The exercise involved dissemination of the Spanish Society for Pulmonology and Thoracic Surgery (SEPAR) guidelines. The use of a portable device to perform spirometry tests was evaluated to adequately categorize COPD and reduce other diagnostic interventions. METHODS: A representative sample of GPs from general practices in Spain was recruited for the study. In phase I, GPs performed an initial evaluation of 5 patients with COPD. In phase II, GPs were randomly allocated to the following groups: 1) control group-G1 (GPs managing COPD according to usual clinical practice); 2) training group-G2 (dissemination of SEPAR guidelines); and 3) training group-G3 (dissemination of SEPAR guidelines and distribution of the Koko Peak Pro to measure FEV(1), FEV(6), and FEV(1)/FEV(6)). RESULTS: Phase-I included 3254 physicians, who selected 16,024 patients. In phase II, 301 physicians in G1, 1182 in G2, and 1141 in G3 selected 1481, 5798, and 5556 patients respectively. Evaluation of the changes in COPD stratification according to the SEPAR guidelines showed that physicians in G1 adequately classified 60% of patients, compared to 69% in G2 and 88.5% in G3 (p<0.0001). On comparing groups G1 and G3, a significant reduction was seen in chest X-rays (from 42% to 32%, p=0.0002) and arterial blood gas studies performed (from 34% to 22%, p<0.0001). CONCLUSIONS: The dissemination of the SEPAR guidelines and the utilization of the portable device for spirometry may improve management of COPD in primary care.

An economic analysis of pharmacological treatment of COPD in Spain.

BACKGROUND: COPD is a prevalent disease that generates high use of resources. The objective of this study was to quantify the economic consequences of non-adherence to GOLD guidelines for the management of COPD patients. METHOD: An economic model was generated to compare different scenarios of observed vs. expected costs of COPD treatment. A pooled analysis of data derived from a systematic review of studies describing treatment of COPD in Spain was combined with drug costs (using different assumptions) to obtain the observed cost of COPD treatment. An expected cost was obtained with the minimum and maximum treatment intensity derived from the GOLD recommendations. RESULTS: A total of 8 studies were identified, comprising 6339 patients. Average medication cost of COPD patients was estimated as being between euro 1218 and euro 1314 per patient per year, higher than the ideal expected average cost (between euro 1007 and euro 1021 per patient/year). Thus, implementation of guidelines would result in a mean reduction of euro 198-euro 293 per patient/year. Sensitivity analysis showed that about 13% of patients had higher treatment costs than the maximum expected cost. This proportion is much higher in moderately/severely affected patients than in mildly affected patients (28.0% and 11.1%, respectively). CONCLUSIONS: Treatment of COPD allows for the identification of areas of inefficiency. An improvement in the adherence to the GOLD guidelines would imply potential savings of medication costs of about 20% of the observed costs.

Health-related quality of life in outpatients with COPD in daily practice: the VICE Spanish Study.

BACKGROUND: The objective of this study was to measure health-related quality of life (HRQL) in outpatients with chronic obstructive pulmonary disease (COPD) and to assess differences in HRQL according to age, gender, and severity of COPD. METHODS: A total of 9405 patients (79% men, mean age 68 years) participated in a cross-sectional study. HRQL was measured with the Short Form 12 Health Survey Questionnaire (SF-12). Severity of COPD was graded into three levels according to forced expiratory volume in one second value. RESULTS: COPD severity was mild in 33.8% of cases, moderate in 49.3% and severe in 16.8%. The mean physical component summary (PCS-12) and mental component summary (MCS-12) scores were 36.8 +/- 10.4 and 47.2 +/- 11.2, respectively. General health and physical functioning domains were those with the lowest scores. The mean MCS-12 scores were significantly higher in men (47.9 +/- 10.9) than in women (44.1 +/- 11.8) (P < 0.001). Patients older than 60 years rated HRQL worse than patients aged 40-59 years. There were statistically significant differences according to severity of disease in the mean scores of all domains of the PCS-12 and MCS-12 scales. CONCLUSIONS: The present findings show the influence of female gender, older age and moderate-to-severe of airflow limitation on HRQL in outpatients with COPD attended in daily practice.

Use of spirometry and patterns of prescribing in COPD in primary care.

OBJECTIVE: To investigate the use and interpretation of spirometry in primary care (PC) in the diagnosis of chronic obstructive pulmonary disease (COPD) and to identify the treatment schedules administered. METHODS: An observational study was performed in a randomized sample of 251 PC physicians including 2130 patients with COPD. Data on the performance of spirometry and the results and the treatment administered were collected as were sociodemographic and clinical data. RESULTS: Spirometric results were obtained in 1243 (58.4%). Most (1118/1243; 89.9%) corresponded to FEV1 (%) values with a mean of 57% (SD=21.5%). It is of note that only 31.8% of spirometric results provided post-bonchodilator results, and 42.9% and 43.1% of the spirometries presented not plausible FVC or FEV1 values, respectively. Treatment varied greatly, with more than 3 drugs being prescribed in 30.6% of the cases. Long-acting beta-2 agonists and inhaled corticosteroids were prescribed in more than 50% of the patients. Tiotropium was administered in 32.4%. According to the GOLD guidelines, 22.8% of the patients in GOLD II, 50% in III and 66.7% in IV were receiving incorrect treatment. CONCLUSIONS: Only 58.4% of the cases included had undergone spirometry. Important deficiencies were observed in the interpretation of the results of spirometry. These difficulties may influence the low implementation of treatment guidelines in COPD in PC.

[Impact of chronic obstructive pulmonary disease on activities of daily living: results of the EIME multicenter study]. / Impacto de la EPOC en la vida diaria de los pacientes. Resultados del estudio multicéntrico EIME.

OBJECTIVE: The aim of this study was to determine the impact of chronic obstructive pulmonary disease (COPD) on activities of daily living in a large group of patients in Spain who responded to a specific questionnaire. A second aim was to explore the practical utility of the questionnaire and determine which variables could be used to identify "fragile" patients or patients in greater need of attention. To do this, we examined the relationship between questionnaire results and clinical variables, lung function measurements, socioeconomic status, and validated quality of life questionnaires. METHODS: We conducted an observational, descriptive, multicenter, cross-sectional study in which 227 respiratory specialists from all over Spain collected data from 1057 patients with COPD. Each patient was given a specific questionnaire containing 7 items that measured the extent to which COPD affected different aspects of their lives. The patients rated each item on a scale of 0 to 2, depending on the level of impact. Total possible scores, thus, ranged from 0 to 14, and patients with a score of 9 or higher were classified as fragile. We then explored the relationship between questionnaire results and clinical variables, socioeconomic status, spirometric values, and quality of life as measured by the St George's Respiratory Questionnaire (SGRQ). RESULTS: We studied 1057 patients (95.2% male) with a mean (SD) age of 67 (9) years and a mean predicted forced expiratory volume in 1 second (FEV1) of 41.8% (13.3%). The mean questionnaire score was 6.3 (3.1). The activities that were affected most were sport and leisure, habitual physical activity, and sex life (major impact reported by 52.5%, 30.3%, and 20.2% of patients, respectively). We found a correlation between questionnaire scores and known disease severity markers such as SGRQ scores, dyspnea, number of exacerbations, and FEV1 in liters. Patients included in the fragile category were older and had a lower socioeconomic status. CONCLUSIONS: COPD impact questionnaire scores correlated well with SGRQ scores and the usual clinical variables and lung function measurements for evaluating disease severity (dyspnea, FEV1, and exacerbations). The questionnaire could, therefore, be a useful tool for identifying fragile patients who, in addition to having poorer clinical status and lung function measurements, have a lower socioeconomic status.

[Use of spirometry in the diagnosis and treatment of chronic obstructive pulmonary disease in primary care]. / Utilización de la espirometría en el diagnóstico y tratamiento de la EPOC en atención primaria.

OBJECTIVE: The aim of this study was to assess the use of spirometry for the diagnosis and follow-up of patients with chronic obstructive pulmonary disease (COPD) in primary care in terms of deficiencies and the requirements for its correct use, and to identify the regimens most commonly used in patients with COPD. METHODS: The study included 839 primary care physicians, each of whom completed 2 questionnaires, one on treatment of COPD and the other on the use of spirometry for diagnosis and follow-up of the disease. RESULTS: Notable among the results was the high number of questionnaires in which no response was given to the question on classification of patients according to the severity of airway obstruction (10.7% of cases) and the low number of correct responses to questions on treatment with bronchodilators during the stable phase of COPD (15.1%). The highest rate of correct responses was for questions regarding the indication for spirometry, all of which were answered correctly in more than 60% of cases. Only 59.2% of primary health care centers performed spirometry, mainly due to a lack of training. In more than 30% of cases the nursing staff had not received specific training, a finding that was reflected in the poor compliance with guidelines for calibration (10.9% of health care centers performed daily calibrations), cleaning of the spirometer (in 13.9% of cases the equipment was never cleaned), and providing patients with pretest recommendations (30% did not provide recommendations the day before spirometry). CONCLUSIONS: Primary care physicians are aware of the usefulness of spirometry for the diagnosis and follow-up of COPD. Although they are able to recognize airflow obstruction, they do not classify patients correctly in terms of severity. Very limited availability of spirometry was observed in primary health care centers and there was little training in the use of the technique, a finding reflected in the poor compliance with guidelines for its use.

Utilización de la espirometría en el diagnóstico y tratamiento de la EPOC en atención primaria / Use of spirometry in the diagnosis and treatment of chronic obstructive pulmonary disease in primary care

Objetivo: El objetivo del estudio ha sido evaluar las deficiencias y necesidades para la correcta utilización de la espirometría en el diagnóstico y seguimiento del paciente con enfermedad pulmonar obstructiva crónica (EPOC) en atención primaria (AP) y conocer las pautas habituales de tratamiento de esta enfermedad. Métodos: Participaron 839 médicos de AP y cada uno de ellos cumplimentó 2 cuestionarios, uno sobre el tratamiento de la EPOC y otro de utilización de la espirometría en su diagnóstico y seguimiento. Resultados: Destacó el bajo índice tanto de respuestas a la pregunta sobre la clasificación de los pacientes en función de la gravedad de la obstrucción (no respondió el 10,7%) como de respuestas correctas en las preguntas sobre el tratamiento broncodilatador en fase estable (respuestas correctas: 15,1%). Las mayores tasas de respuestas correctas se obtuvieron en las preguntas referentes a la indicación de la espirometría, todas ellas con un índice de acierto superior al 60%. Sólo un 59,2% de los centros de AP realizaban espirometrías, sobre todo debido a la falta de formación. En más de un 30% de los casos el personal de enfermería no había recibido formación específica, lo que se reflejaba en un escaso seguimiento de las normativas en cuanto a calibración (un 10,9% de los centros la realizaba diariamente), limpieza de los aparatos (un 13,9% no la hacía nunca) y recomendaciones al paciente (un 30% no daba recomendaciones el día antes). Conclusiones: Los médicos de AP conocen la utilidad de la espirometría en el diagnóstico y seguimiento de la EPOC. Identifican la presencia de una obstrucción al flujo aéreo, pero no se clasifica correctamente a los pacientes en función de su gravedad. Se ha observado una escasa disponibilidad de la espirometría en los centros de AP, así como una escasa formación en su manejo, lo que se refleja en un escaso seguimiento de las normativas de realización de la prueba

Objective: The aim of this study was to assess the use of spirometry for the diagnosis and follow-up of patients with chronic obstructive pulmonary disease (COPD) in primary care in terms of deficiencies and the requirements for its correct use, and to identify the regimens most commonly used in patients with COPD. Methods: The study included 839 primary care physicians, each of whom completed 2 questionnaires, one on treatment of COPD and the other on the use of spirometry for diagnosis and follow-up of the disease. Results: Notable among the results was the high number of questionnaires in which no response was given to the question on classification of patients according to the severity of airway obstruction (10.7% of cases) and the low number of correct responses to questions on treatment with bronchodilators during the stable phase of COPD (15.1%). The highest rate of correct responses was for questions regarding the indication for spirometry, all of which were answered correctly in more than 60% of cases. Only 59.2% of primary health care centers performed spirometry, mainly due to a lack of training. In more than 30% of cases the nursing staff had not received specific training, a finding that was reflected in the poor compliance with guidelines for calibration (10.9% of health care centers performed daily calibrations), cleaning of the spirometer (in 13.9% of cases the equipment was never cleaned), and providing patients with pretest recommendations (30% did not provide recommendations the day before spirometry). Conclusions: Primary care physicians are aware of the usefulness of spirometry for the diagnosis and follow-up of COPD. Although they are able to recognize airflow obstruction, they do not classify patients correctly in terms of severity. Very limited availability of spirometry was observed in primary health care centers and there was little training in the use of the technique, a finding reflected in the poor compliance with guidelines for its use

Chronic respiratory symptoms, spirometry and knowledge of COPD among general population.

RATIONALE: Infradiagnosis of chronic obstructive pulmonary disease (COPD) may be related to the lack of knowledge about the disease and/or the scarce use of diagnostic procedures. This study analyses the frequency of respiratory symptoms and the knowledge about COPD in the general population, together with the use of spirometry in individuals at risk of COPD. POPULATION AND METHOD: A telephone survey was carried out in 6758 subjects older than 40 years, stratified by age, habitat (urban or rural) and region, screened by random-digit dialling. RESULTS: Up to 24% reported having at least one chronic respiratory symptom and 20.9% had a self-reported respiratory diagnosis. A total of 19.2% were active smokers and 40% had never tried to quit. Only 60% of the individuals with chronic symptoms had consulted a physician and, of them, only 45% had undergone spirometry. Spirometry was mentioned more frequently by subjects attended by pulmonologists than by GPs (67.6 vs. 28.6%; P<0.001). The term COPD was identified only by 8.6% of the participants. CONCLUSIONS: Many individuals with respiratory symptoms do not request medical attention and do not attempt to quit smoking. There is a lack of knowledge about COPD. Physicians should more actively inform about the disease and increase the use of spirometry for early detection.

Exacerbations, hospital admissions and impaired health status in chronic obstructive pulmonary disease.

Impaired health status may be a risk factor for frequent exacerbations and hospital admission and, in turn, exacerbations and admissions may further impair the health status of patients with COPD. We have investigated the variables associated with frequent exacerbations (3 or more per year) and admission, with particular interest in health status, in a cohort of ambulatory patients with moderate to severe COPD attended by chest physicians in Spain. A total of 227 investigators included 1057 patients with a mean predicted FEV1 of 41.8%. The mean total score on the St. George's Respiratory Questionnaire (SGRQ) was 47.9 units, and 300 patients (28.4%) had 3 or more exacerbations the previous year and 344 (32.6%) were admitted at least once during the same period. In multivariate analysis only the SGRQ total score was significantly associated with both frequent exacerbations (OR = 1.04; 95% CI = 1.01-2.12; p<0.0001) and admission (OR = 1.01; 95% CI = 1.00-1.02; p = 0.0008). Other variables significantly associated with frequent exacerbations were chronic mucus hypersecretion, increased baseline dyspnea, comorbidity, GOLD stage IV and treatment with inhaled corticosteroids. Variables associated with admission were lower educational level, increased number of exacerbations, use of long-term oxygen therapy and use of short acting beta-2 agonists. An impaired health status is associated with an increased number of exacerbations and with admission to hospital in the previous year. These results, together with previous studies, confirm that impairment in health status in COPD is both a marker of risk for, and a consequence of, frequent exacerbations and admissions.