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HFE gene mutation and oxidative damage biomarkers in patients with myelodysplastic syndromes and its relation to transfusional iron overload: an observational cross-sectional study.

De Souza GF, Ribeiro HL, De Sousa JC, Heredia FF, De Freitas RM, Martins MR, Gonçalves RP, Pinheiro RF, Magalhães SM - BMJ Open (2015)

Bottom Line: Serum MDA levels were highest in patients with MDS with IOL, with a significant difference when compared with patients without IOL and healthy volunteers, pointing to the relationship between IOL and oxidative stress.The GPx and SOD were also significantly higher in these patients, indicating that lipid peroxidation increase was followed by an increase in antioxidant capacity.We observed a significant increase in MDA levels in patients with MDS and IOL, suggesting an increased lipid peroxidation in these patients.

View Article: PubMed Central - PubMed

Affiliation: Post-Graduate Program in Medical Science, Department of Clinical Medicine, Federal University of Ceara, Fortaleza, Ceara, Brazil Cancer Cytogenomic Laboratory, Federal University of Ceara, Fortaleza, Ceara, Brazil.

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Related in: MedlinePlus

PCR-RFLP (restriction fragment length polymorphism) blots of the C282Y, H63D and S65C mutations. Six patients are represented. M: DNA marker 100bp (Invitrogen). Band sizes are indicated on the left and right of the panel. Homozygous allele of C282Y (fragments 296bp, 116bp, 29bp); heterozygous allele of C282Y (fragments 296bp, 145bp, 116bp, 29bp); homozygous allele of H63D (only fragment of 496bp); heterozygous allele of H63D (fragments 496bp, 352bp and 144bp). Wild allele S65C (fragments of 274bp, 147bp, 69bp and 6bp).
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BMJOPEN2014006048F1: PCR-RFLP (restriction fragment length polymorphism) blots of the C282Y, H63D and S65C mutations. Six patients are represented. M: DNA marker 100bp (Invitrogen). Band sizes are indicated on the left and right of the panel. Homozygous allele of C282Y (fragments 296bp, 116bp, 29bp); heterozygous allele of C282Y (fragments 296bp, 145bp, 116bp, 29bp); homozygous allele of H63D (only fragment of 496bp); heterozygous allele of H63D (fragments 496bp, 352bp and 144bp). Wild allele S65C (fragments of 274bp, 147bp, 69bp and 6bp).

Mentions: Genomic DNA was extracted from peripheral blood using the method described by Biometrix Diagnostica/DNA Biopur. HFE genotyping for the C282Y, H63D and S65C mutations was performed using the PCR/restriction fragment length polymorphism (PCR-RFLP). The protocol was the same as that described by Feder11 and Simonsen.12 Fragments of 296bp and 145bp for the wild alleles (282CC), of 296bp, 116bp and 29bp for the homozygous allele (282YY) and of 296bp, 145bp, 116bp and 29bp for the heterozygous allele (282CY) were used for identification. Only one fragment of 496bp was used for the homozygous allele (63DD) and 496bp, 352bp and 144bp were used for the heterozygous allele (63HD). The fragments 352bp and 144bp corresponded to the wild allele (63HH). The fragments of 274bp, 147bp, 69bp and 6bp were used to detect the wild allele (65SS), the fragments 274bp, 216bp, 147bp, 69bp and 6bp for the heterozygous allele (65SC) and the fragments 274bp, 216bp and 6bp for the allele mutant (65CC) (figure 1).


HFE gene mutation and oxidative damage biomarkers in patients with myelodysplastic syndromes and its relation to transfusional iron overload: an observational cross-sectional study.

De Souza GF, Ribeiro HL, De Sousa JC, Heredia FF, De Freitas RM, Martins MR, Gonçalves RP, Pinheiro RF, Magalhães SM - BMJ Open (2015)

PCR-RFLP (restriction fragment length polymorphism) blots of the C282Y, H63D and S65C mutations. Six patients are represented. M: DNA marker 100bp (Invitrogen). Band sizes are indicated on the left and right of the panel. Homozygous allele of C282Y (fragments 296bp, 116bp, 29bp); heterozygous allele of C282Y (fragments 296bp, 145bp, 116bp, 29bp); homozygous allele of H63D (only fragment of 496bp); heterozygous allele of H63D (fragments 496bp, 352bp and 144bp). Wild allele S65C (fragments of 274bp, 147bp, 69bp and 6bp).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4390728&req=5

BMJOPEN2014006048F1: PCR-RFLP (restriction fragment length polymorphism) blots of the C282Y, H63D and S65C mutations. Six patients are represented. M: DNA marker 100bp (Invitrogen). Band sizes are indicated on the left and right of the panel. Homozygous allele of C282Y (fragments 296bp, 116bp, 29bp); heterozygous allele of C282Y (fragments 296bp, 145bp, 116bp, 29bp); homozygous allele of H63D (only fragment of 496bp); heterozygous allele of H63D (fragments 496bp, 352bp and 144bp). Wild allele S65C (fragments of 274bp, 147bp, 69bp and 6bp).
Mentions: Genomic DNA was extracted from peripheral blood using the method described by Biometrix Diagnostica/DNA Biopur. HFE genotyping for the C282Y, H63D and S65C mutations was performed using the PCR/restriction fragment length polymorphism (PCR-RFLP). The protocol was the same as that described by Feder11 and Simonsen.12 Fragments of 296bp and 145bp for the wild alleles (282CC), of 296bp, 116bp and 29bp for the homozygous allele (282YY) and of 296bp, 145bp, 116bp and 29bp for the heterozygous allele (282CY) were used for identification. Only one fragment of 496bp was used for the homozygous allele (63DD) and 496bp, 352bp and 144bp were used for the heterozygous allele (63HD). The fragments 352bp and 144bp corresponded to the wild allele (63HH). The fragments of 274bp, 147bp, 69bp and 6bp were used to detect the wild allele (65SS), the fragments 274bp, 216bp, 147bp, 69bp and 6bp for the heterozygous allele (65SC) and the fragments 274bp, 216bp and 6bp for the allele mutant (65CC) (figure 1).

Bottom Line: Serum MDA levels were highest in patients with MDS with IOL, with a significant difference when compared with patients without IOL and healthy volunteers, pointing to the relationship between IOL and oxidative stress.The GPx and SOD were also significantly higher in these patients, indicating that lipid peroxidation increase was followed by an increase in antioxidant capacity.We observed a significant increase in MDA levels in patients with MDS and IOL, suggesting an increased lipid peroxidation in these patients.

View Article: PubMed Central - PubMed

Affiliation: Post-Graduate Program in Medical Science, Department of Clinical Medicine, Federal University of Ceara, Fortaleza, Ceara, Brazil Cancer Cytogenomic Laboratory, Federal University of Ceara, Fortaleza, Ceara, Brazil.

Show MeSH
Related in: MedlinePlus