Application to quantify fetal lung branching on rat explants

Recently, regulating mechanisms of branching morphogenesis of fetal lung rat explants have been an essential tool for molecular research. The development of accurate and reliable segmentation techniques may be essential to improve research outcomes. This work presents an image processing method to measure the perimeter and area of lung branches on fetal rat explants. The algorithm starts by reducing the noise corrupting the image with a pre-processing stage. The outcome is input to a watershed operation that automatically segments the image into primitive regions. Then, an image pixel is selected within the lung explant epithelial, allowing a region growing between neighbouring watershed regions. This growing process is controlled by a statistical distribution of each region. When compared with manual segmentation, the results show the same tendency for lung development. High similarities were harder to obtain in the last two days of culture, due to the increased number of peripheral airway buds and complexity of lung architecture. However, using semiautomatic measurements, the standard deviation was lower and the results between independent researchers were more coherent

Automated image analysis of lung branching morphogenesis from microscopic images of fetal rat explants

Background: Regulating mechanisms of branching morphogenesis of fetal lung rat explants have been an essential tool for molecular research. This work presents a new methodology to accurately quantify the epithelial, outer contour and peripheral airway buds of lung explants during cellular development from microscopic images. Methods: The outer contour was defined using an adaptive and multi-scale threshold algorithm whose level was automatically calculated based on an entropy maximization criterion. The inner lung epithelial was defined by a clustering procedure that groups small image regions according to the minimum description length principle and local statistical properties. Finally, the number of peripheral buds were counted as the skeleton branched ends from a skeletonized image of the lung inner epithelial. Results: The time for lung branching morphometric analysis was reduced in 98% in contrast to the manual method. Best results were obtained in the first two days of cellular development, with lesser standard deviations. Non-significant differences were found between the automatic and manual results in all culture days. Conclusions: The proposed method introduces a series of advantages related to its intuitive use and accuracy, making the technique suitable to images with different lightning characteristics and allowing a reliable comparison between different researchers.

Automated image analysis of lung branching morphogenesis

Regulating mechanisms of branchingmorphogenesis of fetal lung rat explants have been an essential tool formolecular research.This work presents a new methodology to accurately quantify the epithelial, outer contour, and peripheral airway buds of lung explants during cellular development frommicroscopic images. Methods.Theouter contour was defined using an adaptive and multiscale threshold algorithm whose level was automatically calculated based on an entropy maximization criterion. The inner lung epithelium was defined by a clustering procedure that groups small image regions according to the minimum description length principle and local statistical properties. Finally, the number of peripheral buds was counted as the skeleton branched ends from a skeletonized image of the lung inner epithelia. Results. The time for lung branching morphometric analysis was reduced in 98% in contrast to themanualmethod. Best results were obtained in the first two days of cellular development, with lesser standard deviations. Nonsignificant differences were found between the automatic and manual results in all culture days. Conclusions. The proposed method introduces a series of advantages related to its intuitive use and accuracy, making the technique suitable to images with different lighting characteristics and allowing a reliable comparison between different researchers.

Application to quantify fetal lung branching on rat explant

Recently, regulating mechanisms of branching morphogenesis of fetal lung rat explants have been an essential tool for molecular research. The development of accurate and reliable segmentation techniques may be essential to improve research outcomes. This work presents an image processing method to measure the perimeter and area of lung branches on fetal rat explants. The algorithm starts by reducing the noise corrupting the image with a pre-processing stage. The outcome is input to a watershed operation that automatically segments the image into primitive regions. Then, an image pixel is selected within the lung explant epithelial, allowing a region growing between neighbouring watershed regions. This growing process is controlled by a statistical distribution of each region. When compared with manual segmentation, the results show the same tendency for lung development. High similarities were harder to obtain in the last two days of culture, due to the increased number of peripheral airway buds and complexity of lung architecture. However, using semiautomatic measurements, the standard deviation was lower and the results between independent researchers were more coherent.

Tratamento radiocirúrgico de metástases cerebrais: a relação da histologia, localização cerebral e sintomas

Objetivo – Caracterizar clínica e estatisticamente os doentes com metástases cerebrais submetidos a radiocirurgia. Metodologia – Análise retrospetiva dos doentes com metástases cerebrais submetidos a radiocirurgia com Linac no Hospital do Meixoeiro, sendo a informação analisada no SPSS, versão 18. Resultados – Avaliaram‑se 116 doentes com metástases cerebrais. As localizações primárias de pulmão (54,30%) e mama (21,60%) predominaram. Destacaram‑se como sintomas mais frequentes: cefaleias, fraqueza motora, hemiparesia, paresia e tonturas. Confirma‑se a existência de correlação entre os sintomas decorrentes da presença de metástase e a sua localização cerebral, evidenciando a sua importância no diagnóstico precoce das metástases. O lobo frontal foi a localização cerebral predominante. Discussão e Considerações Finais – Verifica‑se que tendencialmente não existe correlação entre a localização primária e a localização cerebral da metástase. O número de metástases tratadas não sugere ter influência no tempo de sobrevida após o seu diagnóstico. A realização de cirurgia e/ou administração de radioterapia holocraniana previamente à radiocirurgia não apresentou prolongamento de sobrevida em comparação com os doentes não submetidos a tratamento prévio. ABSTRACT: Objective – To characterize clinically and statistically patients with brain metastases who underwent radiosurgery. Methodology – Retrospective analysis of patients with brain metastases that underwent linear accelerator‑based radiosurgery in Hospital do Meixoeiro, and the information analyzed in SPSS version 18. Results – Were evaluated 116 patients with brain metastases. Primary tumors of lung (54.30%) and breast (21.60%) were predominant. Symptoms that stood out as common: headache, motor deficit, hemiparesis, paresis and dizziness. It was confirmed the existence of a correlation between the symptoms arising from the presence of metastasis and its brain location, showing its importance in early diagnosis of metastases. The frontal lobe and the parietal lobe represented the most affected locations by brain metastases. Discussion of results and Concluding Remarks – It verified that tends to be no correlation between the primary location of the tumor and the location of brain metastasis. The number of treated metastases didn’t suggest influence on survival after their diagnosis. The realization of surgery and/or administration of whole‑brain irradiation therapy prior to radiosurgery, showed no prolongation of survival compared with patients that were not submitted to previous treatment.

Prevalência da doença pulmonar obstrutiva crónica em Lisboa, Portugal: estudo burden of obstructive lung disease

Introdução - A prevalência da doença pulmonar obstrutiva crónica (DPOC) apresenta valores muito heterogéneos em todo o mundo. A iniciativa Burden of Obstructive Lung Disease (BOLD) foi desenvolvida para que a prevalência da DPOC possa ser avaliada com metodologia uniformizada. O objetivo deste estudo foi estimar a prevalência da DPOC em adultos com 40 ou mais anos numa população alvo de 2 700 000 habitantes na região de Lisboa, de acordo com o protocolo BOLD. Métodos - A amostra foi estratificada de forma aleatória multifaseada selecionando-se 12 freguesias. O inquérito compreendia um questionário com informação sobre fatores de risco para a DPOC e doença respiratória autoreportada; adicionalmente, foi efetuada espirometria com prova de broncodilatação. Resultados - Foram incluídos 710 participantes com questionário e espirometria aceitáveis. A prevalência estimada da DPOC na população no estadio GOLD I+ foi de 14,2% (IC 95%: 11,1; 18,1) e de 7,3% no estadio ii+ (IC 95%: 4,7; 11,3). A prevalência não ajustada foi de 20,2% (IC 95%: 17,4; 23,3) no estadio i+ e de 9,5% (IC 95%: 7,6; 11,9) no estadio ii+. A prevalência da DPOC no estadio GOLD II+ aumentou com a idade, sendo mais elevada no sexo masculino. A prevalência estimada da DPOC no estadio GOLD I+ foi de 9,2% (IC 95%: 5,9; 14,0) nos não fumadores versus 27,4% (IC 95%: 18,5; 38,5) nos fumadores com carga tabágica de ≥ 20 Unidades Maço Ano. Detetou-se uma fraca concordância entre a referência a diagnóstico médico prévio e o diagnóstico espirométrico, com 86,8% de subdiagnósticos. Conclusões - O achado de uma prevalência estimada da DPOC de 14,2% sugere que esta é uma doença comum na região de Lisboa, contudo com uma elevada proporção de subdiagnósticos. Estes dados apontam para a necessidade de aumentar o grau de conhecimento dos profissionais de saúde sobre a DPOC, bem como a necessidade de maior utilização da espirometria nos cuidados de saúde primários.

Comparação entre o pencil beam convolution algorithm e o analytical anisotropic algorithm em tumores de mama

Mestrado em Radioterapia

The predominance of M2-polarized macrophages in the stroma of low-hypoxic bladder tumors is associated with BCG immunotherapy failure

OBJECTIVE: Bacillus Calmette-Guérin (BCG) immunotherapy is the gold standard treatment for superficial bladder tumors with intermediate/high risk of recurrence or progression. However, approximately 30% of patients fail to respond to the treatment. Effective BCG therapy needs precise activation of the type 1 helper cells immune pathway. Tumor-associated macrophages (TAMs) often assume an immunoregulatory M2 phenotype and may directly interfere with the BCG-induced antitumor immune response. Thus, we aim to clarify the influence of TAMs, in particular of the M2 phenotype in stroma and tumor areas, in BCG treatment outcome. PATIENTS AND METHODS: The study included 99 patients with bladder cancer treated with BCG. Tumors resected before treatment were evaluated using immunohistochemistry for CD68 and CD163 antigens, which identify a lineage macrophage marker and a M2-polarized specific cell surface receptor, respectively. CD68+ and CD163+ macrophages were evaluated within the stroma and tumor areas, and high density of infiltrating cells spots were selected for counting. Hypoxia, an event known to modulate macrophage phenotype, was also assessed through hypoxia induced factor (HIF)-1α expression. RESULTS: Patients in whom BCG failed had high stroma-predominant CD163+ macrophage counts (high stroma but low tumor CD163+ macrophages counts) when compared with the ones with a successful treatment (71% vs. 47%, P = 0.017). Furthermore, patients presenting this phenotype showed decreased recurrence-free survival (log rank, P = 0.008) and a clear 2-fold increased risk of BCG treatment failure was observed in univariate analysis (hazard ratio = 2.343; 95% CI: 1.197-4.587; P = 0.013). Even when adjusted for potential confounders, such as age and therapeutic scheme, multivariate analysis revealed 2.6-fold increased risk of recurrence (hazard ratio = 2.627; 95% CI: 1.340-5.150; P = 0.005). High stroma-predominant CD163+ macrophage counts were also associated with low expression of HIF-1α in tumor areas, whereas high counts of CD163+ in the tumor presented high expression of HIF-1α in tumor nests. CONCLUSIONS: TAMs evaluation using CD163 is a good indicator of BCG treatment failure. Moreover, elevated infiltration of CD163+ macrophages, predominantly in stroma areas but not in the tumor, may be a useful indicator of BCG treatment outcome, possibly owing to its immunosuppressive phenotype.

Modeling of the lung impedance using a fractional order ladder network with constant phase elements

The self similar branching arrangement of the airways makes the respiratory system an ideal candidate for the application of fractional calculus theory. The fractal geometry is typically characterized by a recurrent structure. This study investigates the identification of a model for the respiratory tree by means of its electrical equivalent based on intrinsic morphology. Measurements were obtained from seven volunteers, in terms of their respiratory impedance by means of its complex representation for frequencies below 5 Hz. A parametric modeling is then applied to the complex valued data points. Since at low-frequency range the inertance is negligible, each airway branch is modeled by using gamma cell resistance and capacitance, the latter having a fractional-order constant phase element (CPE), which is identified from measurements. In addition, the complex impedance is also approximated by means of a model consisting of a lumped series resistance and a lumped fractional-order capacitance. The results reveal that both models characterize the data well, whereas the averaged CPE values are supraunitary and subunitary for the ladder network and the lumped model, respectively.

Budget impact from the incorporation of positron emission tomography – computed tomography for staging lung cancers

OBJECTIVE To estimate the budget impact from the incorporation of positron emission tomography (PET) in mediastinal and distant staging of non-small cell lung cancer.METHODS The estimates were calculated by the epidemiological method for years 2014 to 2018. Nation-wide data were used about the incidence; data on distribution of the disease´s prevalence and on the technologies’ accuracy were from the literature; data regarding involved costs were taken from a micro-costing study and from Brazilian Unified Health System (SUS) database. Two strategies for using PET were analyzed: the offer to all newly-diagnosed patients, and the restricted offer to the ones who had negative results in previous computed tomography (CT) exams. Univariate and extreme scenarios sensitivity analyses were conducted to evaluate the influence from sources of uncertainties in the parameters used.RESULTS The incorporation of PET-CT in SUS would imply the need for additional resources of 158.1 BRL (98.2 USD) million for the restricted offer and 202.7 BRL (125.9 USD) million for the inclusive offer in five years, with a difference of 44.6 BRL (27.7 USD) million between the two offer strategies within that period. In absolute terms, the total budget impact from its incorporation in SUS, in five years, would be 555 BRL (345 USD) and 600 BRL (372.8 USD) million, respectively. The costs from the PET-CT procedure were the most influential parameter in the results. In the most optimistic scenario, the additional budget impact would be reduced to 86.9 BRL (54 USD) and 103.8 BRL (64.5 USD) million, considering PET-CT for negative CT and PET-CT for all, respectively.CONCLUSIONS The incorporation of PET in the clinical staging of non-small cell lung cancer seems to be financially feasible considering the high budget of the Brazilian Ministry of Health. The potential reduction in the number of unnecessary surgeries may cause the available resources to be more efficiently allocated.

Evaluation of lung function abnormalities prevalence in poultry workers

Introduction - Poultry workers can be at an increased risk of occupational respiratory diseases, like asthma, chronic obstructive pulmonary disease and extrinsic allergic alveolitis. Spirometry screening is fundamental to early diagnosis trough the identification of related ventilatory defects. Purpose - We aimed to assess the prevalence of lung function abnormalities in poultry workers.

Characterisation of adventitious lung sounds in people with dementia

Introduction: Lower Respiratory Tract Infections (LRTIs) are highly prevalent in institutionalised people with dementia, constituting an important cause of morbidity and mortality. Computerised auscultation of Adventitious Lung Sounds (ALS) has shown to be objective and reliable to assess and monitor respiratory diseases, however its application in people with dementia is unknown. Aim: This study characterised ALS (crackles and wheezes) in institutionalised people with dementia. Methods: An exploratory descriptive study, including 6 long-term care institutions was conducted. The sample included a dementia group (DG) of 30 people with dementia and a match healthy group (HG) of 30 elderly people. Socio-demographic and anthropometric data, cognition, type and severity of dementia, cardio-respiratory parameters, balance, mobility and activities and participation were collected. Lung sounds were recorded with a digital stethoscope following Computerised Respiratory Sound Analysis (CORSA) guidelines. Crackles’ location, number (N), frequency (F), two-cycle duration (2CD), initial deflection width (IDW) and largest deflection width (LDW) and wheezes’ number (N), ratio (R) and frequency (F) were analysed per breathing phase. Statistical analyses were performed using PASW Statistics(v.19). Results: There were no significant differences between the two groups in relation to the mean N of crackles during inspiration and expiration in both trachea and thorax. DG trachea crackles had significant higher F during inspiration and lower IDW, 2CD and LDW during expiration when compared with HG. At the thorax, the LDW during inspiration was also significantly lower in the DG. A significant higher N of inspiratory wheezes was found in the HG. Both groups had a low ratio of high frequency wheezes. Conclusion: Computerised analyses of ALS informed on the respiratory system and function of people with dementia and elderly people. Hence, this could be the step towards prevention, early diagnosis and continuous monitoring of respiratory diseases in people with cognitive impairment.

Inapparent lung involvement in patients with the subacute juvenile type of paracoccidioidomycosis

Three patients with the diagnosis of subacute juvenile paracoccidioidomycosis who, at the time of their first visit, had no signs or symptoms of lung involvement, were studied. Initially the diagnosis was confirmed by the observation of P. brasiliensis in biopsy material obtained from clinically involved lymphadenopathies. The lung X-rays done in all patients, did not reveal pathologic changes, although it was possible to observe and isolate the fungus from sputum samples obtained from the three patients. This fact reinforces the pulmonary genesis of the mycosis and proofs the existence of a pulmonary primary infection, even in patients with the juvenile manifestations, in whom the lung component is obscured by the predominant lymph node involvement.