PMID- 10648790
OWN - NLM
STAT- MEDLINE
DCOM- 20000328
LR  - 20231105
IS  - 1362-4962 (Electronic)
IS  - 0305-1048 (Print)
IS  - 0305-1048 (Linking)
VI  - 28
IP  - 4
DP  - 2000 Feb 15
TI  - A catalytic antioxidant metalloporphyrin blocks hydrogen peroxide-induced 
      mitochondrial DNA damage.
PG  - 968-73
AB  - Reactive oxygen species (ROS) have been implicated as the cause of cumulative 
      damage to DNA, proteins and lipids that can ultimately result in cell death. A 
      common problem when measuring oxidative DNA damage has been the introduction of 
      modifications in the native state of the molecule by many DNA isolation methods. 
      We circumvented this problem by employing direct PCR (DPCR) of whole cell 
      lysates. DPCR of mouse lung fibroblasts performed better than PCRs containing 
      template acquired by phenol/chloroform extraction or a commercially available 
      genomic DNA isolation kit. We investigated the direct use of whole cell 
      preparations in the polymerase chain reaction (PCR) to detect hydrogen peroxide 
      (H(2)O(2))-mediated DNA damage. We observed a concentration-dependent decrease in 
      amplification efficiency of a 4.3 kb mitochondrial (mt)DNA target in 
      H(2)O(2)-treated mouse lung fibroblasts (MLFs). At low doses the efficiency of 
      amplification returns to control levels over 24 h. We detected no change in 
      amplification efficiency of a plasmid control containing our mtDNA target under 
      any of the culture conditions employed in these studies. Treatment of MLFs with 
      the catalytic antioxidant manganese(III) meso -tetrakis(4-benzoic acid)porphyrin 
      (MnTBAP) attenuates the effects of H(2)O(2)exposure. When quantitated with an 
      external standard the use of DPCR in tandem with a PCR amplification efficiency 
      assay provides a powerful approach to assess oxidative mtDNA damage.
FAU - Milano, J
AU  - Milano J
AD  - Department of Medicine, Room K706, Goodman Building, National Jewish Research 
      Center, 1400 Jackson Street, Denver, CO 80206, USA.
FAU - Day, B J
AU  - Day BJ
LA  - eng
GR  - R01HL59602/HL/NHLBI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - England
TA  - Nucleic Acids Res
JT  - Nucleic acids research
JID - 0411011
RN  - 0 (Antioxidants)
RN  - 0 (DNA Primers)
RN  - 0 (DNA, Mitochondrial)
RN  - 0 (Metalloporphyrins)
RN  - BBX060AN9V (Hydrogen Peroxide)
SB  - IM
MH  - Animals
MH  - Antioxidants/*pharmacology
MH  - Base Sequence
MH  - Catalysis
MH  - *DNA Damage
MH  - DNA Primers
MH  - DNA, Mitochondrial/drug effects
MH  - Hydrogen Peroxide/*antagonists & inhibitors
MH  - Male
MH  - Metalloporphyrins/*pharmacology
MH  - Mice
MH  - Mice, Inbred C57BL
MH  - Mitochondria/*drug effects/metabolism
MH  - Polymerase Chain Reaction
PMC - PMC102572
EDAT- 2000/01/29 00:00
MHDA- 2000/04/01 00:00
PMCR- 2000/02/15
CRDT- 2000/01/29 00:00
PHST- 2000/01/29 00:00 [pubmed]
PHST- 2000/04/01 00:00 [medline]
PHST- 2000/01/29 00:00 [entrez]
PHST- 2000/02/15 00:00 [pmc-release]
AID - gkd189 [pii]
AID - 10.1093/nar/28.4.968 [doi]
PST - ppublish
SO  - Nucleic Acids Res. 2000 Feb 15;28(4):968-73. doi: 10.1093/nar/28.4.968.