A fibrose cística (FC) é diagnosticada na presença de achados fenotípicos, história familiar ou triagem neonatal positiva acompanhada de evidência laboratorial de disfunção da CFTR, seja pelo teste do suor, diferença de potencial nasal ou pela identificação de duas mutações conhecidas como causa de FC nos genes da CFTR. Objetivos: Avaliar a contribuição da análise molecular do gene CFTR na investigação diagnóstica da fibrose cística em pacientes com suspeita de FC leve ou doença atípica. Secundariamente, comparar as características dos pacientes em 3 grupos: grupo com identificação de duas mutações conhecidas como causadoras da FC, grupo com identificação de apenas uma mutação e grupo sem mutação identificada. Métodos: Estudo transversal em adolescentes e adultos (≥14 anos). Os pacientes foram submetidos à avaliação clínica, laboratorial e radiológica; espirometria, microbiologia do escarro, ecografia hepática, teste do suor e análise molecular do gene CFTR. Resultados: Foram avaliados 37 pacientes com achados fenotípicos de FC, com ou sem confirmação pelo teste do suor. Houve predomínio do sexo feminino (75,7%) com média de idade de 32,5 ± 13,6 anos. A análise molecular contribuiu para o diagnóstico definitivo de FC em 3 casos (8,1%) dentre 37 pacientes em avaliação. Em 7 pacientes (18,9%) foram identificadas apenas uma mutação causadora de FC e em 26 pacientes (70,3%) não foram identificadas mutações. Nenhuma característica clínica estudada se associou com o diagnóstico genético. A mutação p.F508del foi a mais comum, encontrada em 5 pacientes. A associação de p.V232D e p.F508del foi encontrada em 2 pacientes. Outras mutações encontradas foram: p.A559T, p.D1152H, p.T1057A, p.I148T, p.V754M, p.P1290P e p.R1066H e p.T351S. Conclusão: A análise molecular da região codificante do gene CFTR apresentou contribuição limitada para investigação diagnóstica de pacientes com suspeita de fibrose cística leve ou doença atípica. Além disso, não houve associação entre as características clínicas e o diagnóstico genético. / Cystic fibrosis (CF) is diagnosed in the presence of phenotypic findings, family history or positive neonatal screening accompanied by laboratory evidence of CFTR dysfunction, either by sweat test, nasal potential difference or the identification of two mutations known to cause CF in the CFTR gene. Objectives: To evaluate the contribution of molecular analysis of CFTR gene in cystic fibrosis diagnostic investigation in patients with suspected mild FC or atypical disease. Secondarily, to compare the characteristics of patients into 3 groups: group with identification of two mutations known to cause CF, group with identification of just one mutation and group without mutations. Methods: Cross-sectional study in adolescent and adult (≥ 14 years). The patient underwent clinical, laboratory and radiological spirometry, sputum microbiology, liver ultrasound, sweat test and molecular analysis of the CFTR gene. Results: We evaluated 37 patients with phenotypic findings of FC, with or without confirmation by the sweat test. There was a predominance of females (75.7%) with a mean age of 32.5 ± 13.6 years. Molecular analysis contributed to the definitive diagnosis of CF in 3 cases (8.1%) among 37 patients under evaluation. In 7 patients (18.9%) were identified only one mutation that causes CF and in 26 patients (70.3%) were not identified mutations. No clinical feature studied was associated with genetic diagnosis. The P.F508del mutation was the most common, found in 5 patients. The association p.V232D and p.F508del was found in 2 patients. Other mutations found were: p.A559T, p.D1152H, p.T1057A, p.I148T, p.V754M, and p.P1290P p.R1066H and p.T351S. Conclusion: Molecular analysis of the CFTR gene coding region showed limited contribution to the diagnostic investigation of patients with suspected cystic fibrosis mild or atypical disease. Moreover, there was no association between clinical features and genetic diagnosis.

A fibrose cística (FC) é diagnosticada na presença de achados fenotípicos, história familiar ou triagem neonatal positiva acompanhada de evidência laboratorial de disfunção da CFTR, seja pelo teste do suor, diferença de potencial nasal ou pela identificação de duas mutações conhecidas como causa de FC nos genes da CFTR. Objetivos: Avaliar a contribuição da análise molecular do gene CFTR na investigação diagnóstica da fibrose cística em pacientes com suspeita de FC leve ou doença atípica. Secundariamente, comparar as características dos pacientes em 3 grupos: grupo com identificação de duas mutações conhecidas como causadoras da FC, grupo com identificação de apenas uma mutação e grupo sem mutação identificada. Métodos: Estudo transversal em adolescentes e adultos (≥14 anos). Os pacientes foram submetidos à avaliação clínica, laboratorial e radiológica; espirometria, microbiologia do escarro, ecografia hepática, teste do suor e análise molecular do gene CFTR. Resultados: Foram avaliados 37 pacientes com achados fenotípicos de FC, com ou sem confirmação pelo teste do suor. Houve predomínio do sexo feminino (75,7%) com média de idade de 32,5 ± 13,6 anos. A análise molecular contribuiu para o diagnóstico definitivo de FC em 3 casos (8,1%) dentre 37 pacientes em avaliação. Em 7 pacientes (18,9%) foram identificadas apenas uma mutação causadora de FC e em 26 pacientes (70,3%) não foram identificadas mutações. Nenhuma característica clínica estudada se associou com o diagnóstico genético. A mutação p.F508del foi a mais comum, encontrada em 5 pacientes. A associação de p.V232D e p.F508del foi encontrada em 2 pacientes. Outras mutações encontradas foram: p.A559T, p.D1152H, p.T1057A, p.I148T, p.V754M, p.P1290P e p.R1066H e p.T351S. Conclusão: A análise molecular da região codificante do gene CFTR apresentou contribuição limitada para investigação diagnóstica de pacientes com suspeita de fibrose cística leve ou doença atípica. Além disso, não houve associação entre as características clínicas e o diagnóstico genético. / Cystic fibrosis (CF) is diagnosed in the presence of phenotypic findings, family history or positive neonatal screening accompanied by laboratory evidence of CFTR dysfunction, either by sweat test, nasal potential difference or the identification of two mutations known to cause CF in the CFTR gene. Objectives: To evaluate the contribution of molecular analysis of CFTR gene in cystic fibrosis diagnostic investigation in patients with suspected mild FC or atypical disease. Secondarily, to compare the characteristics of patients into 3 groups: group with identification of two mutations known to cause CF, group with identification of just one mutation and group without mutations. Methods: Cross-sectional study in adolescent and adult (≥ 14 years). The patient underwent clinical, laboratory and radiological spirometry, sputum microbiology, liver ultrasound, sweat test and molecular analysis of the CFTR gene. Results: We evaluated 37 patients with phenotypic findings of FC, with or without confirmation by the sweat test. There was a predominance of females (75.7%) with a mean age of 32.5 ± 13.6 years. Molecular analysis contributed to the definitive diagnosis of CF in 3 cases (8.1%) among 37 patients under evaluation. In 7 patients (18.9%) were identified only one mutation that causes CF and in 26 patients (70.3%) were not identified mutations. No clinical feature studied was associated with genetic diagnosis. The P.F508del mutation was the most common, found in 5 patients. The association p.V232D and p.F508del was found in 2 patients. Other mutations found were: p.A559T, p.D1152H, p.T1057A, p.I148T, p.V754M, and p.P1290P p.R1066H and p.T351S. Conclusion: Molecular analysis of the CFTR gene coding region showed limited contribution to the diagnostic investigation of patients with suspected cystic fibrosis mild or atypical disease. Moreover, there was no association between clinical features and genetic diagnosis.

Trata-se de uma pesquisa exploratória descritiva com abordagem qualitativa cujo objetivo foi conhecer como a criança de seis a doze anos que vive com fibrose cística percebe seu corpo no cotidiano. O contexto foi a residência das crianças, que moram em Porto Alegre e Região Metropolitana. Os participantes do estudo foram cinco crianças atendidas na Associação de Apoio a Portadores de Mucoviscidose do Rio Grande do Sul (AMUCORS) com idades entre seis e doze anos que vivem com fibrose cística, sendo a seleção realizada de forma intencional, por meio da indicação da presidente da associação. A coleta das informações ocorreu por meio de fotografia associada com entrevista no domicílio. Para realização da coleta foram realizadas três visitas domiciliares, após um primeiro contato propiciado pela AMUCORS. Na primeira visita ocorreu a apresentação à criança e ao responsável da proposta do estudo e a obtenção do seu aceite e do seu responsável. Nesse encontro também foi entregue uma máquina fotográfica para a criança e solicitado que fizesse registros fotográficos tentando responder a questão do estudo. Na segunda visita era recolhida a máquina para posterior impressão das fotos e agendado o próximo encontro. Na terceira visita, com as fotos impressas, ocorreu a entrevista a partir das fotos selecionadas pelo participante. A análise das informações foi realizada de acordo com a análise de conteúdo proposta por Minayo. Os aspectos éticos da pesquisa foram respeitados, sendo o projeto aprovado pela Comissão de Pesquisa da Escola de Enfermagem da UFRGS (COMPESQ/EE) e pelo Comitê de Pesquisa e Ética da UFRGS. Os temas emergidos na interpretação das informações foram: cuidados diários com a doença, atividades habituais modificadas para a faixa etária, lazer prejudicado e fé e esperança. Destaca-se que, para a maioria das crianças do estudo, o cotidiano de medicamentos e fisioterapia está incorporado no seu modo de ser no cotidiano e parece não acarretar muitas frustrações. Nesse sentido, a criança com fibrose cística não percebe seu corpo diferente de um corpo saudável e, sim, sentem que necessitam de mais cuidados para poder realizar as atividades correspondentes a sua faixa etária. A ampla divulgação dos resultados de estudos como esse poderá contribuir para o aprimoramento da prática do cuidado na área da saúde e, em especial para a enfermagem, visando melhorar a qualidade de vida dessas crianças. Enfatiza-se ainda, o uso de máquina fotográfica como estratégia de coleta de informações em pesquisa qualitativa com crianças. / This is an exploratory descriptive qualitative approach aims to better understand how the six to twelve years living with cystic fibrosis realizes her body in everyday. The context was the residence of the children, who live in Porto Alegre and Metropolitan Area. Study participants were five children attending the Association for Supporting People with Cystic Fibrosis of Rio Grande do Sul (AMUCORS) aged between six and twelve living with cystic fibrosis, the selection being made intentionally, by indicating the president of the association. Data collection occurred through photography associated with household interview. To collect de information were three visits, after a first contact propitiated by AMUCORS. The first visit took place the presentation of children and responsibility of the proposed study and obtains their acceptance and their responsibility. This meeting was also given a camera for the child and asked him made photographic records trying to answer the question of the study. On the second visit was collected the machine for later print of the photos and the next scheduled meeting. On the third visit, with photos printed, the interview took place from the pictures selected by the participant. Analysis of the data was performed according to content analysis proposed by Minayo. The ethical aspects of the research were respected, and the project is approved by the Research Committee of the School of Nursing at UFRGS (COMPESQ/EE) and the Research and Ethics Committee of UFRGS. The themes that emerged in the interpretation of information were: daily care with the disease, usual activities for age, leisure harmed and faith and hope. It is noteworthy that, for most children in the study, the daily medication and physiotherapy is embedded in their way of being in everyday and does not seem to cause much frustration. In this sense, the child with cystic fibrosis does not realize his body other than a healthy body, and yes, they feel they need more care to be able to carry out the activities corresponding to their age group. The wide dissemination of the results of such studies can contribute to improving the care practice in health and in particular for nursing, to improve the quality of life of these children. It also emphasizes the use of camera as a strategy for collecting information in qualitative research with children. / Se trata de un enfoque cualitativo descriptivo exploratorio con el objetivo de comprender mejor cómo los niños de seis a doce años que viven con fibrosis quística se da cuenta de su cuerpo en el diario. El contexto fue la residencia de los hijos, que viven en Porto Alegre y el Área Metropolitana. Los participantes del estudio fueron cinco niños que asisten a la Asociación de Apoyo a Personas con Fibrosis Quística de Rio Grande do Sul ( AMUCORS ) de edades comprendidas entre los seis y los doce años que viven con fibrosis quística, la selección se hizo intencionadamente, indicando el presidente de la asociación . Los datos fueron recolectados a través de fotografías asociados con la entrevista en el hogar. Para recopilar la información de tres visitas, después de un primer contacto proporcionada por los AMUCORS. La primera visita tuvo lugar la presentación de los niños y la responsabilidad del estudio propuesto y obtener su aceptación y su responsabilidad. Esta reunión también se le entregó una cámara para el niño y le pidió que hiciera los registros fotográficos que tratan de responder a la pregunta del estudio. En la segunda visita se recogió la máquina para su posterior impresión de fotos y programada la próxima reunión. En la tercera visita, con fotos impresas, la entrevista se llevó a cabo a partir de las fotografías seleccionadas por el participante. El análisis de los datos se realizó de acuerdo al análisis de contenido propuesto por MINAYO. Se respetarán los aspectos éticos de la investigación, y el proyecto es aprobado por el Comité de Investigación de la Escuela de Enfermería de la UFRGS (COMPESQ / EE) y el Comité de Investigación y Ética de la UFRGS. Los temas que surgieron en la interpretación de la información fueron: el cuidado diario con la enfermedad, las actividades normales modificado para la edad, el ocio discapacitado y la fe y la esperanza. Es de destacar que, para la mayoría de los niños en el estudio, la medicación diaria y la fisioterapia está incrustado en su forma de ser en el diario y no parece causar mucha frustración. En este sentido, el niño con fibrosis quística no se da cuenta que su cuerpo no sea un cuerpo sano, y sí, ellos sienten que necesitan más atención para poder llevar a cabo las actividades correspondientes a su grupo de edad. La amplia difusión de los resultados de tales estudios puede contribuir a mejorar la práctica de la atención en salud y en particular para enfermería, para mejorar la calidad de vida de estos niños. El énfasis es seguir utilizando la cámara como una estrategia para la recopilación de información en la investigación cualitativa con los niños.

Orientador: José Dirceu Ribeiro / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Made available in DSpace on 2018-08-21T08:23:59Z (GMT). No. of bitstreams: 1 Aguiar_KatiaCristinaAlberto_M.pdf: 5558123 bytes, checksum: 9ecbadfc1ac6fd35012e0cfd9b4cfeff (MD5) Previous issue date: 2012 / Resumo: Introdução: As características emocionais da Fibrose Cistica na adolescência, na visão pessoal de quem a vivencia é um tema pouco referenciado na literatura científica. A adolescência é um período por si próprio, carregado de possibilidades de conflitos emocionais. A associação com uma doença crônica e seu manejo, nesta etapa da vida pode maximizar estes conflitos. Objetivo: O presente estudo teve como objetivo compreender como é para o adolescente vivenciar a fibrose cística (FC), doença que necessita tratamento diário por toda vida e participação ativa do próprio paciente. A pesquisa estudou como os adolescentes convivem com a doença, quais suas necessidades e como eles percebem o futuro. Método: Utilizou-se o método clinico qualitativo, empregando-se a técnica de entrevista semi estruturada de questões abertas. Quarenta e dois pacientes aceitaram participar do estudo. Suas falas foram divididas em oito categorias e dez subcategorias que descrevem suas respostas em relação á vivência com a FC. Resultados: Os resultados evidenciam que os adolescentes apresentam sentimentos de medo da morte, vergonha, raiva, perda da liberdade, da escola, dos amigos e da confiança em si mesmo, necessidade de ajustamento ás rotinas diferenciadas, necessidade de igualdade e aceitação, perspectiva negativa e positiva em relação ao futuro e expectativas em relação ao futuro com FC. Conclusões: Os sentimentos relatados são influenciados pela doença e dificultam a maneira como eles lidam com ela. Espera-se que a compreensão dessas vivências possa estimular estudos longitudinais nos centros que atendem pacientes com FC / Abstract: Introduction: The emotional characteristics of a chronic disease in adolescence, in a personal vision of those who experience it, is a topic not much referenced in the scientific literature. Adolescence by itself is a life period full of emotional conflict possibilities. The association with a chronic disease and how to cope with it at this specific stage of life can increase these conflicts. Objective: The present research goal is to understand how adolescents deal with cystic fibrosis (CF), an illness which needs a diary treatment for a lifetime and active patient participation. It highlighted how adolescents live with the disease, what are their needs and what they expect from the future. Method: A clinical qualitative method was used employing a semi structured interview technique of open questions. A total of forty two patients accept to be involved in the study. Their speeches were divided into eight categories that describe the patient's answers according to how adolescents cope with CF. Results: Results highlighted that adolescents show feelings like fear of death, shame, rage, lost of freedom, school year, friend and self-confidence, needs of adjustment to differentiated routines, needs of equality and acceptance, negative perspective for the future, positive perspective for the future and expectations about the future with CF. Conclusions: Such feelings are influenced by the disease and input barriers on how adolescents cope with it. The comprehension of how to deal with a chronic disease should stimulate longitudinal studies at all centers that assist CF patients / Mestrado / Saude da Criança e do Adolescente / Mestra em Ciências

Class of 2012 Abstract / Specific Aims: The epithelial sodium channels (ENaC) found on the apical membranes of epithelial cells including those lining the respiratory tract are the rate limiting step of the absorption of excess fluid from the airspace of the alveoli. ENaC function is modulated by the effects of various physiologic signals such as the adrenergic and purinergic pathways, in addition to other local channels which control the flow of negatively charged ions such as the cystic fibrosis transmembrane conductance regulator (CFTR). We sought to determine the influence of genetic variation on the alpha subunit of ENaC at amino acid position 663 on baseline exhaled ions and pulmonary function in patients with CF. Methods: We assessed pulmonary function ( forced vital capacity[FVC], forced expiratory volume in one second [FEV1], forced expiratory flow maximum[FEFmax]) using a Medical Graphics cardiopulmonary testing device (Minneapolis, MN). Measures of exhaled sodium (Na+) and chloride (Cl-) were obtained using exhaled breathe condensate collected on a Jaeger Ecoscreen condenser unit (Cardinal Health, Yorba Linda, CA) with Na+ quantification using an atomic absorption spectrophotometer (Analyst 100; Perkin Elmer, Norwalk, CT) and Cl- anion quantification using a Dionex AS11 HC column. Healthy n=31 (n=18[58%], 9[29%], and 4[13%] subjects; Body mass index (BMI)=23±1, 25±2, and 25±2kg/ m2 for AA, AT and TT groups respectively). CF n= 42 (n=33[79%], 7[16%], and 2[5%] subjects; BMI equals 23±7, 19±0.4, and 20±2.2kg/m2 for AA, AT and TT groups respectively). Main Results: We found that the distribution of genotypes in CF differed from healthy subjects, with the AA genotype in 80% of CF and 59% in healthy. No significant difference were demonstrated in healthy subjects between genotype groups for pulmonary function and exhaled chloride while the genotypes did differ in exhaled Na (Na=2.9±0.4, 1.7±0.3, and 3.7±1.1mmol/L for AA, AT, and TT respectively, ANOVA p=0.07). CF subjects with the AA genotype had a higher baseline exhaled Cl-, FEV1, and FEFmax than those in the AA group (Cl=0.125±0.038,0.0 27±0.007, and 0.033±0.02 mmol/L ; FEV1=71±5, 68±11, and 40±22L; FEFmax=86±4, 72±7, and 44±24L/sec; for AA, AT, and TT respectively, ANOVA p<0.05, Tukey [AA vs. TT] p<0.05) while exhaled Na+ and FVC were similar between genotypes. Conclusions: Our results suggest that CF subjects with the AA genotype of the alpha subunit of the ENaC have a higher baseline exhaled Cl- and a resulting increase in pulmonary function when compared to the overactive TT groupCF patients with the TT αENaC genotype are likely candidates for early identification and treatment with inhaled ENaC inhibitors or other modulators of this pathway in order to improve survival.

Fibrose du tissu adipeux humain: relations avec l'élasticité du tissu, des changements de poids et les comorbiditiés de l'obésité.Contexte Le remodelage de la matrice extracellulaire (MEC), e.g. la fibrose, est une altération pathologique du tissu adipeux sous-cutané (TAsc) dans l'obésité. La fibrose du TAsc est associée aux troubles métaboliques et à la perte de poids à la chirurgie bariatrique (CB). Un outil non-invasif a été développé pour évaluer l'élasticité du TAsc (la VS) et son lien avec les comorbidités de l'obésité.Hypothèses 1) La VS associée aux troubles métaboliques prédit la perte de poids et les améliorations cardiométaboliques à la CB; 2) La VS est liée avec les collagènes et avec d'autres caractères du TAsc; 3) la VS change avec le remodelage de la MEC du TAsc durant la perte de poids; 4) le développement de l'outil évaluant la VS est utile dans la pratique clinique. Méthodes Une étude clinque a été réalisée chez plus de 100 obèses candidats à la CB, avec les mesures transcriptomique, histologique et secretomique in vitro et ex vivo pour évaluer des altérations du TAsc. La validation de l'outil a été aussi réalisée. Résultats 1) La VS est associée à la fibrose du TAsc, aux paramètres biocliniques, aux anomalies cardiométaboliques chez les obèses morbides; 2) La VS pré-CB prédit la perte de poids d'un an à la CB; 3) Le TAsc subit un remodelage majeur durant la perte de poids, en particulier une augmentation des collagènes qui est lié à une élévation de la dégradation des collagènes, à une diminution du pontage de la MEC, non à un changement détectable de la VS. Ces observations suggèrent une adaptation adéquate de la MEC du TAsc durant la perte de poids. Conlusion : L’exploration du remodelage du TAsc (e.g. la fibrose et la VS) est un intérêt principal dans cette maladie complexe. / Context Extracellular matrix (ECM) remodeling, e.g. fibrosis, is a hallmark pathological alteration of subcutaneous adipose tissue (scAT) in obesity. ScAT fibrosis associates with metabolic disorders and bariatric surgery (BS)-induced weight loss. ScAT fibrosis increases mechanical stress on adipocytes and induces fibro-inflammation. A non-invasive tool was recently developed to evaluate scAT stiffness, shear wave speed (SWS) and its link with obesity comorbidities. Hypotheses 1) SWS associates with metabolic disorders and predicts BS-induced weight loss and cardiometabolic improvements, 2) SWS is explained by collagen deposition and other structural features; 3) SWS changes with scAT ECM remodeling during weight loss; 4) developing a relevant tool to measure SWS is helpful in clinical practice.Methods We conducted a clinical study in more than 100 obese candidates for BS, combined with a series of in vitro and ex vivo transcriptomic, histological and secretomic measures of tissue alteration. Validation steps were realized to confirm the clinical use of the tool. Results 1) SWS associated with scAT fibrosis, various bioclinical parameters, cardiometabolic abnormalities at baseline in morbid obesity; 2) pre-BS SWS has a predictive role in one-year BS-induced weight loss; 3) scAT underwent major remodeling, particularly collagens accumulation, during BS-induced weight loss. This was related to increased collagen degradation, decreased cross-linking, but not with detectable change in SWS. These observations reflected an adequate ECM adaptation during fat mass loss. Conclusion Exploring scAT remodeling (e.g. fibrosis and stiffness) is of main interest in this complex disease.

A descriptive correlational study investigated the possible relationship between family functioning and the self-concept development of adolescents with cystic fibrosis (CF). Twenty-two adolescents ranging from 13 to 19 years of age and members of their families volunteered to participate. The adolescents completed the Offer Self-image Questionnaire (OSIQ) and the Family Assessment Device (FAD). Thirty-four family members completed the FAD. Mean standard scores for the study population were compared to normative values for the OSIQ and the FAD. The Spearman rho correlation procedure was used to investigate relationships between scales of the two measures. Findings for the self-concept measure (OSIQ) revealed that the mean scores for the adolescents with CF were better than normative values with two exceptions; the mean score was lower than normative values on the Sexual Attitudes Scale for males and females ranging from 13 to 15 years of age and on the Body and Self-image Scale for males 13 to 19 years and females 13 to 15 years of age. On the family functioning measure (FAD) the mean scores for adolescents and their family members were lower than suggested healthy cut off scores (Epstein, Baldwin, & Bishop, 1983) with the exception of a higher score on the Roles Scale. However, scores of the sample were similar to FAD scores generated from a random sample considered by the authors of the FAD to be descriptive of the general family population (Miller, Bishop, Epstein, & Keitner, 1985). A positive relationship between well developed adolescent self-concept and positive family functioning was not indicated; most correlations between the OSIQ and FAD scales were negative. Health care for adolescents with CF should include addressing their sexual and body and self-image concerns and promoting healthy family functioning. Further investigation of self-concept and family functioning for adolescents with CF is warranted. / Applied Science, Faculty of / Nursing, School of / Graduate

<p>Radiation-induced lung injury (RILI) is a common condition in the setting of lung and breast cancer. Often, patients who suffer from RILI experience pneumonitis and pulmonary fibrosis months after treatment. These pathologies have commonly been modeled using mice and observing their deterioration until mortality and quantifying pathology on histological sections.<b> </b></p> <p>With this study, we used a longitudinal microCT and a 7T MRI to characterize male C57Bl/6 mice irradiated with a single dose of 20 Gy to the whole thoracic area delivered by an X-Rad cabinent irradiator. CT was performed with a respiratory gating sequence at 2 week timepoints to construct an RIPF model. The fraction of RIPF to total lung volume was calculated at each time point from images, and the data was anaylzed using one-way ANOVA Welch and Dunnett’s T3 multiple comparisons tests. Tidal lung volumes were also calculated and anlyazed in a simlar manner. Mice were then imaged using MRI and CT at 0, 5, and 8 week timepoints to compare results. These results were analyzed for comparison (ANOVA and Dunnett’s T3) and correlation (Pearson’s r) with each other. Histology was later performed using H&E and Trichrome stains to provide ex-vivo verification of pathology.<b> </b>At the 10-12 week time point ( ) significant RIPF formed. Weeks proceeding showed increased significance until the 22+ week timepoint, which showed less statistical significance ( ) due to increased variance at this timepoint. Dunnett’s T3 test showed no significant differences between tidal lung volumes over time. Tests also showed no significant differences between CT and MRI results with a correlation coefficient of .<b> </b>Early in the study, problems arose when pre-marture mortality was occurring to a significant portion of our subjects. Analysis later showed issues during irradiation that resulted in significant dose being absorbed by the stomach. Adjusting our shiedling lead to increased early survival of our subjects enabling us to contine our study. Significant RIPF development was not significant until 10-12 weeks post-irradiation, then RIPF became more severe at proceeding timepoints. Tidal lung volume showed no significant deviation over the development of RIPF. This result is most likely affected by the variation of results at later timepoints, since several mice with severe RIPF were significantly hindered in their ability to breathe during the study. MRI results showed close correlation with CT results and prodcued similar values at early timepoints. However, noticeable differences were seen at later timepoints when significant RIPF developed ( ). <b></b></p>

Indiana University-Purdue University Indianapolis (IUPUI) / The Gram-negative bacterium Pseudomonas aeruginosa is an opportunistic pathogen which readily establishes itself in the lungs of people with cystic fibrosis (CF). Most CF patients have life-long P. aeruginosa infections. By modulating its own virulence and forming biofilms, P. aeruginosa is able to evade both host immune responses and antibiotic treatments. Previous studies have shown that the magnesium transporter MgtE plays a role in virulence modulation by inhibiting transcription of the type III secretion system, a mechanism by which bacteria inject toxins directly into the eukaryotic host cell. MgtE had already been identified as a magnesium transporter, and thus its role in regulating cytotoxicity was indicative of dual functions for this protein. This research focused on a structure-function analysis of MgtE, with the hypothesis that the magnesium transport and cytotoxicity functions could be exerted independently. Cytotoxicity assays were conducted using a co-culture model system of cystic fibrosis bronchial epithelial cells and a ∆mgtE strain of P. aeruginosa transformed with plasmids carrying wild type or mutated mgtE. Magnesium transport was assessed using the same mgtE plasmids in a Salmonella strain deficient in all magnesium transporters. Through analysis of a number of mgtE mutants, we found two constructs – a mutation in a putative magnesium binding site, and an N-terminal truncation – which demonstrated a separation of functions. We further demonstrated the uncoupling of functions by showing that different mgtE mutants vary widely in their ability to regulate cytotoxicity, whether or not they are able to transport magnesium. Overall, these results support the hypothesis of MgtE as a dual function protein and may lead to a better understanding of the mechanisms underlying P. aeruginosa virulence. By understanding virulence mechanisms, we may be able to develop treatments to reduce infections and pave the way to better health for people with cystic fibrosis.

Tesis por compendio / [ES] De entre las metodologías disponibles para estudiar la digestión de alimentos, los modelos de digestión in vitro se plantean como procedimientos válidos para este propósito. La digestión in vitro consiste en simular el proceso de digestión en el laboratorio, reproduciendo las condiciones fisiológicas en cuanto a composición de los fluidos digestivos (electrolitos y enzimas), pH, temperatura, fuerzas mecánicas y duración de las etapas oral, gástrica e intestinal. Abordar el estudio de la digestión de nutrientes es de especial relevancia en patologías que cursan con alteraciones pancreáticas o hepáticas, asociadas a una digestión de lípidos comprometida en la etapa intestinal, debido a la disminución de secreción de pancreatina, bicarbonato y sales biliares. Este es el caso de la fibrosis quística con insuficiencia pancreática, y los pacientes que padecen esta afección deben seguir la terapia de sustitución de enzimas pancreáticas, que consiste en el suministro exógeno de pancreatina encapsulada. Sin embargo, la dosis de este suplemento debe ajustarse a las características de los alimentos y no se dispone de ningún método válido para tal fin. Para hacer frente a este reto, en el proyecto financiado con fondos europeos MyCyFAPP se ha logrado desarrollar un método para ajustar la dosis óptima de los suplementos enzimáticos utilizados en la terapia. La presente tesis doctoral se realizó en el marco de dicho proyecto. Concretamente, esta tesis tiene como objetivo abordar el estudio de la digestión de lípidos en los alimentos de origen animal (carne y productos cárnicos, huevos, queso y pescado) en el contexto de la fibrosis quística. Para abordar este objetivo se aplicó un modelo de digestión in vitro estático con el fin de explorar el papel de las características inherentes a los alimentos (estructura de la matriz alimentaria como resultado del procesado) y los factores relacionados con el individuo (pH, concentración de sales biliares y concentración de pancreatina) como factores determinantes de la lipólisis en alimentos de origen animal. A lo largo de los cuatro capítulos presentados, centrados en el huevo, la carne, el queso y el pescado, se presenta un diseño experimental común para estudiar la lipólisis, la proteólisis y la degradación de la matriz. En cada estudio, las diferentes técnicas de procesado aplicadas a los alimentos evaluados también permitieron evaluar el efecto de las propiedades inherentes a los alimentos en los resultados del estudio. Los resultados han contribuido al desarrollo de un nuevo método basado en la evidencia para optimizar la terapia de reemplazo de enzimas pancreáticas e informan a la comunidad científica sobre nuevos conocimientos en el comportamiento de diferentes alimentos sometidos al proceso de digestión. / [CA] De les metodologies disponibles per estudiar la digestió d'aliments, els models de digestió in vitro es plantegen com a procediments vàlids per a aquest propòsit. La digestió in vitro consisteix en simular el procés de digestió al laboratori, reproduint les condicions fisiològiques pel que fa a la composició dels fluids digestius (electròlits i enzims), pH, temperatura, forces mecàniques i durada de les etapes oral, gàstrica i intestinal. Abordar l'estudi de la digestió de nutrients és d'especial rellevància en patologies que cursen amb alteracions pancreàtiques o hepàtiques, associades a una digestió de lípids compromesa en l'etapa intestinal, a causa de la disminució de secreció de pancreatina, bicarbonat i sals biliars. Aquest és el cas de la fibrosi quística amb insuficiència pancreàtica. Els pacients que pateixen aquesta afecció han de seguir la teràpia de substitució d'enzims pancreàtics, que consisteix en el subministrament exogen de pancreatina encapsulada. No obstant això, la dosi d'aquest suplement ha d'ajustar-se a les característiques dels aliments i actualment no es disposa de cap mètode vàlid per a tal fi. Per enfrontar a aquest repte, en el projecte finançat amb fons europeus, MyCyFAPP, s'ha aconseguit desenvolupar un mètode per a ajustar la dosi òptima dels suplements enzimàtics utilitzats en la teràpia. La present tesi doctoral es va realitzar en el marc d'aquest projecte. Concretament, aquesta tesi té com a objectiu abordar l'estudi de la digestió de lípids en els aliments d'origen animal (ous, carn i productes carnis, formatge i peix) en el context de la fibrosi quística. Per a abordar aquest objectiu es va aplicar un model de digestió in vitro estàtic amb la finalitat d'explorar el paper de les característiques inherents als aliments (estructura de la matriu alimentària com a resultat del processament) i els factors relacionats amb l'individu (pH, concentració de sals biliars i concentració de pancreatina) com a factors determinants de la lipòlisi en aliments d'origen animal. Als quatre capítols presentats, centrats en l'ou, carn, formatge i peix, es presenta un disseny experimental comú per a estudiar la lipòlisi, la proteòlisi i la degradació de la matriu. En cada estudi, les diferents tècniques de processament aplicades als aliments avaluats també van permetre avaluar l'efecte de les propietats inherents als aliments en els resultats de l'estudi. Els resultats han contribuït al desenvolupament d'un nou mètode basat en l'evidència per a optimitzar la teràpia de substitució d'enzims pancreàtics i informen la comunitat científica sobre nous coneixements en el comportament de diferents aliments sotmesos al procés de digestió. / [EN] Among the available methodologies to study food digestion, in vitro digestion models have raised as a valid procedure. In vitro digestion consists of simulating the digestion process in the laboratory, by reproducing the physiological conditions in terms of digestive fluids composition (electrolytes and enzymes), pH, temperature, mechanical forces and duration of the oral, gastric and intestinal stages. Addressing the study of nutrient digestion is of special relevance in pathologies coursing with pancreatic or hepatic alterations, which are associated with compromised intestinal lipid digestion due to reduced secretion of pancreatin, bicarbonate and bile salts. This is the case of Cystic Fibrosis along with pancreatic insufficiency, and the patients suffering this condition have to follow pancreatic enzyme replacement therapy, the exogenous supply of encapsulated pancreatin. However, the dose of this supplement should be adjusted to the specific characteristics of foods, and no valid method was available for such purpose. To tackle this challenge, the EU-funded project MyCyFAPP succeed to develop a method to adjust the optimal dose the enzyme supplements used in the therapy. The present doctoral thesis was conducted as a relevant part of this project. Concretely, this thesis aims at addressing the study of lipid digestion in foods to generate new knowledge regarding nutrient digestion in animal origin dietary sources (egg, meat and meat products, cheese and fish) in the context of Cystic Fibrosis. To address this goal a static in vitro digestion model was applied. The role of inherent-to-food characteristics (resulting food matrix structure from processing) and host related factors (pH and bile salts concentration and pancreatin concentration) were explored as determinants of lipolysis in animal-origin foods. Along the four chapters presented, focused on egg, meat, cheese and fish, a common experimental design was applied to study lipolysis, proteolysis and matrix degradation. In each study, different processing techniques applied to the assessed foods allowed for evaluating the effect of inherent-to-food properties on the study outcomes as well. The results have contributed to the development of a new evidence-based method to optimise pancreatic enzyme replacement therapy, and inform the scientific community about new insights on the behaviour of different foods undertaking the digestion process. / Authors of this paper acknowledge the European Union and the Horizon 2020 Research and Innovation Framework Programme (PHC-26-2014 call Self-management of health and disease: citizen engagement and mHealth) for fully funding this research in the context of MyCyFAPP Project, under grant agreement number 643806. The authors would like to thank the Conselleria de Educació i Investigació de la Generalitat Valenciana and also the European Union (“El Fondo Social Europeo (FSE) invierte en tu futuro”) for the PhD scholarship given to Andrea Asensio Grau (ACIF/2017/008). This study was developed thanks to the equipment funded with the support from the Generalitat Valenciana IDIFEDER/2018/041 (PO FEDER Comunitat Valenciana 2014-2020). Finally, we thank Antonio Martínez Cañada, from the Data Science and Biostatistics Unit of Instituto de Investigación Sanitaria La Fe, and Arash Javanidejad for the English corrections. / Asensio Grau, A. (2021). In vitro approach of dietary and host related factors affecting digestion of animal-origin foods under cystic fibrosis disease [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/171512 / Compendio