Seminario Biologia Molecular
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Study of DNA damage caused by dipyrone in presence of some transition metal ions
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Seminario Biologia Molecular
- 1. ALEJANDRO HENAO VILLADA MEDICINE UPB – 2018-1 III SEMESTER PONTIFICAL BOLIVARIAN U N IV E R S IT Y | M E D MOLECULAR BIOLOGY
- 2. Introduction PONTIFICAL BOLIVARIAN U N IV E R S IT Y | M E D MOLECULAR BIOLOGY DNA damage It refers to alterations in the DNA structure caused by physical or chemical agents. If the damage is not repaired, it could cause mutations in genes and then, cell death Dipyrone • It´s a potent analgesic, antipyretic and spasmolytic agent. • The mechanism responsible for the analgesic effect, most probably rests on the inhibition of the COX 3 • Dipyrone also can block both PG- dependent and PG-independent pathways of fever induced by LPS
- 3. Introduction PONTIFICAL BOLIVARIAN U N IV E R S IT Y | M E D MOLECULAR BIOLOGY Some chemical compounds: • L-Dopa • N-acetylcysteine • Gentamicine, etc. Induce DNA damage In presence of transition metal ions Reducing those ions, which in the presence of H2O2 generate free radicals Dipyrone or metamizole is a NSAID that is still prescribed in many countries despite of its dangerous side effects. However, no studies have been made to evaluate the possible DNA damage induced by NaDip in presence of metal ions.
- 4. General objective PONTIFICAL BOLIVARIAN U N IV E R S IT Y | M E D MOLECULAR BIOLOGY To evidentiate in vitro the DNA damage caused by Dipyrone (NaDip) in presence of some transition metal ions which normally, play important roles in a biological system
- 5. Materials and methods PONTIFICAL BOLIVARIAN U N IV E R S IT Y | M E D MOLECULAR BIOLOGY STRATEGY AND PREPARATION OF RECOMBINANT DNA BASIS This concept implies to create deliberately DNA molecules composed by fragments of DNA coming of diferent sources. IT WAS USED FOR… pUC19 is a cloning vector, it means that is used to carry the recombinant DNA from a donor cell to the recipient cell into which the gene to be transferred is inserted.
- 6. Materials and methods PONTIFICAL BOLIVARIAN U N IV E R S IT Y | M E D MOLECULAR BIOLOGY PLASMID DNA EXTRACTION BASIS To extract plasmid DNA, whatever the method used, involves the following steps: 1. Culture of transformed cells 2. Collection and purification of bacteria 3. Purification of the plasmids IT WAS USED FOR… They did not extracted the DNA from a bacteria because they buyed it. The fabricator of the plasmid had to do it in order to produce the artificial plasmids.
- 7. Materials and methods PONTIFICAL BOLIVARIAN U N IV E R S IT Y | M E D MOLECULAR BIOLOGY NUCLEIC ACID ELECTROPHORESIS BASIS It is an analytical technique used to separate DNA or RNA fragments by size and reactivity, in a viscous medium. Nucleic acids (which are negatively charge) migrate to the anode. IT WAS USED FOR… Measuring how much plasmid DNA suffered damage, passing from SC to OC Also it was used for knowing how much smear was formed.
- 8. Results PONTIFICAL BOLIVARIAN U N IV E R S IT Y | M E D MOLECULAR BIOLOGY
- 9. Results PONTIFICAL BOLIVARIAN U N IV E R S IT Y | M E D MOLECULAR BIOLOGY
- 10. Results PONTIFICAL BOLIVARIAN U N IV E R S IT Y | M E D MOLECULAR BIOLOGY
- 11. Results PONTIFICAL BOLIVARIAN U N IV E R S IT Y | M E D MOLECULAR BIOLOGY
- 12. Discussion PONTIFICAL BOLIVARIAN U N IV E R S IT Y | M E D MOLECULAR BIOLOGY AUTHOR ACCORDING TO THEM… IS THIS TRUE HERE? Lloyd and Phillips, 1999 Cu2+ ions are able to interact with DNA in-between the bases Schweigert et al., 2000 DNA-copper-hydroperoxo complexes generates highly reactive species as O2 and •OH Alipázaga et al., 2005 Ni2+ was able of causing pUC19 DNA damage, induced by autoxidation of S(IV), when complexed with GGH (Ni3+ /GGH) with no formation of none Ni+ complexes Iwamoto et al., 2003 DNA damage does not occur in a solution with Mn3+ and AZT even when exposed to UVB
- 13. Conclusions PONTIFICAL BOLIVARIAN U N IV E R S IT Y | M E D MOLECULAR BIOLOGY 1. The effects of Dipyrone or Metamizole should be studied in detail because the authors of this article can not ensure the same behavior of the drug in vivo according to its effects on the DNA structure. 2. Once more, free radicals play an important role in the damage of cellular structures such as DNA, and now in consequense of a drug´s effect with metals. According to this, is important to determine which other drugs could work by the same way.