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Study of Chronic Pulmonary Diseases in Pediatric Patients: bronchiolitis obliterans and cystic fibrosis
Universidad Nacional de Tucuman
Cátedra de Inmunología. Instituto de Microbiología “Dr. Luis C. Verna”. Facultad de Bioquímica, Química, Farmacia y Biotecnología. Universidad Nacional de Tucumán (UNT).
Dr Juan Carlos Valdez
Dra. Gobbato, Nadia
Type of Research Project
- Clinical Project with Laboratory work
What is the background of the project?
The inflammatory response in pediatric patients is studied in a Public Hospital. The clinic, images and laboratory hardly determine the lung damage. Chronic obstructive pulmonary disease, bronchiolitis obliterans and cystic fibrosis are pediatric pulmonary diseases with high morbidity and mortality. The pulmonary deterioration develops as a result of infections and inflammatory response that leads to pulmonary dysfunction. It is necessary to develop new techniques that determine more precisely the inflammatory state of the lungs. In addition to the clinical diagnosis, it is necessary to determine the metabolites of the oxidative stress characteristic of these processes, measuring the fraction exhaled from NO. We are developing the study of the mucus of these patients by nuclear magnetic resonance by determining spectral profiles and metabolites of oxidative stress and relating them to the clinical state. The study of the infecting bacteria and their interaction with polymorphonuclear cells will contribute to a greater knowledge of the pathophysiology of these diseases to project treatments and follow-up forms.
What is the aim of the project?
The general objective of this project is to contribute to the knowledge of pediatric chronic pulmonary infections (Chronic Obstructive Pulmonary Disease, Obliterating Bronchiolitis(BO) and Cystic Fibrosis (CF)) from a scientific perspective that allows to establish the physiopathogenic mechanisms and based on these optimize diagnosis, treatment and follow-up using conventional methods
What techniques and methods are used?
The determination of the pulmonary exacerbation by the exaled fraction of nitric oxide (feNO) comparatively with characteristic profiles and metabolites derived from oxidative stress, using sputum samples (non-invasive method) and relatively inexpensive such as Nuclear Magnetic Resonance, will allow to know more certainty the condition in patients with BO and CF, difficult to detect by other methods. This will allow establishing adequate treatments to improve the evolution of both diseases. The interaction of the infecting bacteria and the polymorphonuclear of the patients will determine whether or not they are prone to develop an exacerbated inflammatory response. The techniques to use are: Determination of exhaled fraction of NO (feNO): a NIOX-MINO (portable electrochemical analyzer) is used, which is the unique breathing measurement method, performed by the doctor in the hospital. Spectral analysis of sputum: by nuclear magnetic resonance that will be analyzed with software. It will be supervised by professionals who work in the Laboratory of Research and Analytical Services (LISA) of the Faculty of Biochemistry, Chemistry Pharmacy. Apoptosis and necrosis: Cell damage will be measured by determination of apoptosis with Annexin V (commercial kit) and necrosis by propidium iodide, by flow cytometry when bacteria isolated from sputum (alone and in co-cultures) interact with polymorphonuclear cells. Polymorphonuclear and Mononuclear cells will be isolated from heparinized circulating blood from healthy volunteers and patients with COPD. NETosis: Induction of Netosis in PMN is performed on polymorphonuclear cells triggered by bacteria using Sytox Green (nuclear dyes). Microbicidal activity: of polymorphonuclear cells on bacteria isolated from sputum. Cellularity of sputum: staining with Giemsa.
What is the role of the student?
- The student will observe the practical experiments but will be highly involved in the analysis of the results
- The tasks will be done under supervision
What are the tasks expected to be accomplished by the student?
All the tasks carried out by the students are supervised by the teachers. The student works under sterile conditions and all safety measures are taken. Students must work before, during and after obtaining clinical samples. They must prepare all the necessary material to carry out the investigations. Students work with clinical samples such as blood and sputum obtained from patients and blood from healthy volunteers, which are obtained when patients attend the Pneumology Service of the Hospital of the Child Jesus. The samples are transported to the faculty laboratory where they separate the blood cells to perform the inflammatory response investigation and the preparation of the sputum samples for determination of cellularity and analysis by nuclear magnetic resonance.
Will there be any theoretical teaching provided (preliminary readings, lectures, courses, seminars etc)
Students receive a very detailed explanation of the research project accompanied by the reading of different authors on the research topic. The results of the laboratory and the clinical studies (carried out by the pneumonologist) are analyzed jointly in order to reach the conclusions that allow us to present the work in national and international congresses.
What is expected from the student at the end of the research exchange? What will be the general outcome of the student?
- The student will prepare a scientific report - No specific outcome is expected
What skills are required of the student? Is there any special knowledge or a certain level of studies needed?
Subject Passed: Basic inmunology and I hope the students are interested in the project
Are there any legal limitations in the student’s involvement
Type of students accepted
This project accepts: - Medical students - Students in biomedical fields
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