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Physiologic and pathologic regulation of the renin angiotensin aldosterone system (RAAS)
Hungary(HuMSIRC)-University of Debrecen, Debrecen
Division of Clinical Physiology, Faculty of Medicine, University of Debrecen
Zoltan Pap MD.
Dr. Miklós Fagyas
Type of Research Project
- Basic science
What is the background of the project?
The renin angiotensin aldosterone system (RAAS) is one of the most important regulator of blood pressure and salt-water homeostasis. Therapeutic inhibition of RAAS is beneficial for the cardiovascular patients. Angiotensin converting enzyme (ACE) and its homologue, ACE2 are members of the RAAS which ensure the balance of tissue angiotensin II concentration. Angiotensin II is the main effector molecule of the RAAS, excess production of angiotensin II may contribute to the development and progression of cardiovascular diseases.
What is the aim of the project?
We hypothesize, that angiotensin II production is altered in cardiovascular diseases in aspect of tissue ACE/ACE2 activity imbalance, lack of endogenous inhibition of ACE etc. We try to find connection between pathological regulation of RAAS and cardiovascular diseases and disease profression.
What techniques and methods are used?
ACE and ACE2 activity measurement with a fluorescent kinetic assay. ACE and 2 concentration measurement with an ELISA technique. Genotyping of RAAS-genes using allele specific PCR technique and gel-electrophoresis. Preparation of samples from blood and human tissues. What is PCR (polymerase chain reaction)? PCR is a technique used in the lab to make millions of copies of a particular section of DNA. • Using PCR it is possible to generate thousands to millions of copies of a particular section of DNA from a very small amount of DNA. How does PCR work? • The principles behind every PCR, whatever the sample of DNA, are the same. • Five core ‘ingredients’ are required to set up a PCR. These are: o the DNA template to be copied o primers, short stretches of DNA that initiate the PCR reaction, designed to bind to either side of the section of DNA you want to copy o DNA bases (A, C, G and T) are the building blocks of DNA and are needed to construct the new strand of DNA o Taq polymerase enzyme to add in the new DNA bases o buffer to ensure the right conditions for the reaction. • PCR involves a process of heating and cooling called thermal cycling which is carried out by machine. • There are three main stages: 1. Denaturing – when the double-stranded template DNA is heated to separate it into two single strands. 2. Annealing – when the temperature is lowered to enable the DNA primers to attach to the template DNA. 3. Extending – when the temperature is raised and the new strand of DNA is made by the Taq polymerase enzyme. • These three stages are repeated 20-40 times, doubling the number of DNA copies each time.
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?
The student has to attain the basic laboratory techniques ( it includes pipetting, preparation of solvents, determining pH, using and adjusting microscopes, basic laboratory calculations) and the basics of measurement assay used in our laboratory, than the student has to perform measurements first with supervision than on his/her own. The student will learn the procedure of analysis and analyze the measured data with different computer programs.
Will there be any theoretical teaching provided (preliminary readings, lectures, courses, seminars etc)
No, but all of the methods and applied techniques will be explained in details.
What is expected from the student at the end of the research exchange? What will be the general outcome of the student?
- The student’s name will be mentioned in a future publication - The student will have the opportunity to present the results together with the supervisor at a conference
What skills are required of the student? Is there any special knowledge or a certain level of studies needed?
knowledge of basic laboratory techniques (pipetting, preparation of solvents, determining pH, using and adjusting microscopes, basic laboratory calculations etc.) basic computer skills (Word, Excel, PowerPoint)
Are there any legal limitations in the student’s involvement
Type of students accepted
This project accepts: - Medical students - Graduated students (less than 6 months)
- (1) Fagyas M. et al: New perspectives in the renin- angiotensin-aldosterone system (RAAS) II: albumin suppresses angiotensin converting enzyme (ACE) activity in human. PLoS One. 2014 Apr 1;9(4):e87844
- Úri K. et al: Circulating ACE2 activity correlates with cardiovascular disease development. J Renin Angiotensin Aldosterone Syst. 2016 Dec 12;17(4).
- Csongrádi et al: Optimized angiotensin- converting enzyme activity assay for the accurate diagnosis of sarcoidosis. Clin Chem Lab Med. 2018 Feb 9.
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