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| | =The Team= | | =The Team= |
| - | [[File:Ufmgteam.jpg|400px|thumb|right|UFMG_Brazil Team]] | + | [[File:Ufmgteam.jpg|400px|thumb|left|UFMG_Brazil Team]] |
| | + | In the beginning, we decided to meet weekly to discuss ideas to be developed in iGEM competition. We scheduled a fixed day and time for such discussion and the presence of all members was considered mandatory, as we had in mind that a good project would require the involvement of everybody. So, based on this perspective and considering that most of our colleagues have a knowhow applied to human health, we thought about tropical diseases, like dengue. We had imagined what we could do to precociously diagnose that disease as a measure to provide a fast and precise health care to patients positively diagnosed. Summarizing, we have firstly considered to develop a fast diagnostic tool for dengue, which everyone could use without restrictions, based on a GMO (Genetically Modified Organism). Despite having made a big effort to implement it, many factors turned it an unviable project. First, we didn’t have means to deal with the vector, Aedes aegypti, and we couldn’t establish a viable way to use a GMO to our primary purposes. To try to solve these problems, we invited a researcher from Funed (Fundação Ezequiel Dias - Brazil), Alzira Batista Cecilio, to talk to us about the disease and the fast diagnostic test that she was developing in her studies. This gave us some possibilities, but all of them were too complex to be applied to iGEM in short time. We kept working on building new ideas to be implemented. So, in order to perform a search, we divided our team in groups, which were encouraged to give new and viable ideas to be developed. One of them had a performable idea: check for biomarkers in order to precociously diagnose heart diseases, a priori based in choline detection. But, as this substance is released to blood flow in response to many disturbs, we thought that more biomarkers would be necessary in order to provide a reliable diagnostic. Occurred to us that it would be interesting to add a biomarker already validated and well described. We thought of using creatine-kinase MB (CK-MB), but it does not have an useable receptor or induciblepromoter available, at least one that we could find, to be expressed on our chassis. Troponin was cogitated to be another of our relevant biomarkers, however, it has also shown to be unviable due to the absence of a receptor we could use and or a channel to transport it into the cell. After a intense search, in the end we have agreed to use three biomarkers: Brain Natriuretic Peptide (BNP), Trimethylamine-N-Oxide (TMAO) and Ischemia Modified Albumin (IMA). BNP is a validated biomarker for Acute Coronary Syndrome (ACS) and it has a receptor that could be used to detect BNP, despite of huge size of its receptor (NPR-A) which have a transmembrane site. TMAO, is not considered a validated biomarker, but it came out in our latest searches it could be used as a heart failure predictor, since this substance attacks the heart muscle tissue and provokes necrosis, the main factor of myocardial infarction. IMA is an indicative of any sort of ischemia and it was validated by FDA as a biomarker for ACS, although it is best used as a negative predictor than a positive one (meaning that its absence indicates that everything is probably fine, but its presence means that there’s something wrong). |
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| | ==Undergraduates== | | ==Undergraduates== |
| | ===Alan Sales Barbosa=== | | ===Alan Sales Barbosa=== |
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| | </div> | | </div> |
| | </html> | | </html> |
| - |
| |
| - | =History=
| |
| - | ==Our history==
| |
| - | In the beginning, we decided to meet weekly to discuss ideas to be developed in iGEM competition. We scheduled a fixed day and time for such discussion and the presence of all members was considered mandatory, as we had in mind that a good project would require the involvement of everybody. So, based on this perspective and considering that most of our colleagues have a knowhow applied to human health, we thought about tropical diseases, like dengue. We had imagined what we could do to precociously diagnose that disease as a measure to provide a fast and precise health care to patients positively diagnosed. Summarizing, we have firstly considered to develop a fast diagnostic tool for dengue, which everyone could use without restrictions, based on a GMO (Genetically Modified Organism). Despite having made a big effort to implement it, many factors turned it an unviable project. First, we didn’t have means to deal with the vector, Aedes aegypti, and we couldn’t establish a viable way to use a GMO to our primary purposes. To try to solve these problems, we invited a researcher from Funed (Fundação Ezequiel Dias - Brazil), Alzira Batista Cecilio, to talk to us about the disease and the fast diagnostic test that she was developing in her studies. This gave us some possibilities, but all of them were too complex to be applied to iGEM in short time. We kept working on building new ideas to be implemented. So, in order to perform a search, we divided our team in groups, which were encouraged to give new and viable ideas to be developed. One of them had a performable idea: check for biomarkers in order to precociously diagnose heart diseases, a priori based in choline detection. But, as this substance is released to blood flow in response to many disturbs, we thought that more biomarkers would be necessary in order to provide a reliable diagnostic. Occurred to us that it would be interesting to add a biomarker already validated and well described. We thought of using creatine-kinase MB (CK-MB), but it does not have an useable receptor or induciblepromoter available, at least one that we could find, to be expressed on our chassis. Troponin was cogitated to be another of our relevant biomarkers, however, it has also shown to be unviable due to the absence of a receptor we could use and or a channel to transport it into the cell. After a intense search, in the end we have agreed to use three biomarkers: Brain Natriuretic Peptide (BNP), Trimethylamine-N-Oxide (TMAO) and Ischemia Modified Albumin (IMA). BNP is a validated biomarker for Acute Coronary Syndrome (ACS) and it has a receptor that could be used to detect BNP, despite of huge size of its receptor (NPR-A) which have a transmembrane site. TMAO, is not considered a validated biomarker, but it came out in our latest searches it could be used as a heart failure predictor, since this substance attacks the heart muscle tissue and provokes necrosis, the main factor of myocardial infarction. IMA is an indicative of any sort of ischemia and it was validated by FDA as a biomarker for ACS, although it is best used as a negative predictor than a positive one (meaning that its absence indicates that everything is probably fine, but its presence means that there’s something wrong).
| |
| - | ==Brainstorming==
| |
| - | ==Day by Day==
| |
| - | ===January 2013===
| |
| - | * Team was formed.
| |
| - | * We decided to have meetings every Tuesday’s afternoon.
| |
| - | * Introductory presentations: what is iGEM, what are Biobricks, computational science.
| |
| - | ===February and March 2013===
| |
| - | * We researched about previous projects developed for iGEM, to give us some direction.
| |
| - | *Each team member had to bring ideas of a project. There were several presentations and discussion until we get to our project: cardiovascular diseases biomarkers.
| |
| - | ===April 2013===
| |
| - | *19th April: we presented our project to professors and students of Biochemistry and Immunology Department. It was very important to test our ability of putting our ideas on a screen and discussing them with people that were not part of the team. It was good to have a feedback.
| |
| - | *At our meetings, we were always studying and discussing about cardiac diseases biomarkers, in order to improve our project.
| |
| - | ===May 2013===
| |
| - | * More improvements to our project.
| |
| - | * Discussions about human practices.
| |
| - |
| |
| - | ===June 2013===
| |
| - | * Final design of our project.
| |
| - | * In late June, we received iGEM’s biobricks kit.
| |
| - |
| |
| - | ===July 2013===
| |
| - | * We started our experiments. We had a lot of trouble, as the chloramphenicol that we were using was expired and didn’t work well (we took a long time to solve this problem...).
| |
| - | * Biosafety Human Practices: a one-week course with Neuza Antunes about biosafety. By the end of the week, an interview was made to conclude the course (colocar link para o vídeo).
| |
| - | * 19th July - Human Practices at UFMG Escolas: it was a wonderful experience! Teaching synthetic biology to children and teenagers was very pleasant. We used our Brickard Game to make it more attractive (colocar links para o arquivo do jogo de cartas e para o vídeo). - Cloning of RCNA+YFP into PSB1A3.
| |
| - |
| |
| - | ===August 2013===
| |
| - | * We finally started fluorimetric assays with the composite RCNA+YFP, using cobalt.
| |
| - | * The oligonucleotide that we had sent to be synthesized (Promoter TorCAD) arrived.
| |
| - | * Cloning of TorCAD into PSB1C3.
| |
| - | * Primers arrived.
| |
| - | * PCRs and digestions to confirm the constructs PSB1A3_RCNA+YFP and PSB1C3_TorCAD.
| |
| - | * 17th August: Jamboré at USP - Meeting of Brazilian iGEM teams.
| |
| - |
| |
| - | ===September 2013===
| |
| - | * More fluorimetric assays.
| |
| - | * Cloning of TorCAD+RFP into PSB1C3.
| |
| - | * We sent our new biobricks to iGEM Headquarters.
| |
| - | * The E. coli Dilemma video (colocar link).
| |
| - | * 21th September: an interview with our team was published in the biggest newspaper of Minas Gerais’ State, “Estado de Minas”.
| |
| - | * 27th September: WIKI FREEZE!!!!
| |
| - |
| |
| - | ===October 2013===
| |
| - | * Regional Jamboree in Chile.
| |
| | | | |
| | =Acknowledgment= | | =Acknowledgment= |
| | {{Team:UFMG Brazil/sponsor}} | | {{Team:UFMG Brazil/sponsor}} |
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