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Methylenetetrahydrofolate reductase C677T variant in Indian children with craniosynostosis: Its role in the pathogenesis, risk of craniosynostosis.

Pandey RK, Ali A, Singh A, Gayan S, Bajpai M - Indian J Hum Genet (2014)

Bottom Line: After successful amplification, a small aliquot (5 μl) of the MTHFR reaction mixture was treated with 1 units of Hinf I restriction enzyme (NEB).Results were obtained and compiled.When comparing the offspring of mothers statistically significant differences were found.

View Article: PubMed Central - PubMed

Affiliation: Department of Pediatric Surgery, All India Institute of Medical Sciences, New Delhi, India.

ABSTRACT

Background: 677C to T allele in the 5, 10-methylenetetrahydrofolate reductase (MTHFR) gene has been implicated in the etiology of various syndromes and nonsyndromic diseases but till date no direct studies have been reported with craniosynostosis.

Objectives: The aim was to study the family-based association of MTHFR polymorphism in different categories of craniosynostosis patients.

Materials and methods: This was a cross-sectional study in which 30 patients classified as Apert syndrome, Pfeiffr syndrome and nonsyndromic craniosynostosis patients with their family were recruited. A sample of 3 ml intravenous blood was taken from patients and from their family members (father and mother) in ethylenediaminetetraacetic acid-anticoagulated vacutainer for the purpose of the study. Genomic DNA was extracted from peripheral blood lymphocytes by phenol chloroform extraction method. Primers for MTHFR gene were designed. The polymerase chain reaction was carried out. After successful amplification, a small aliquot (5 μl) of the MTHFR reaction mixture was treated with 1 units of Hinf I restriction enzyme (NEB). Results were obtained and compiled.

Results: A total of 30 patients/participants with craniosynostosis of Indian descent and their parents formed the study group. The genotyping did not confirm an association between the MTHFR 677C to T polymorphism and between different categories of craniosynostosis. When comparing the offspring of mothers statistically significant differences were found.

Conclusion: C667T polymorphism of the MTHFR gene is unlikely to play a role in the pathogenesis of craniosynostosis though maternal MTHFR C677T polymorphism may be a genetic risk factor.

No MeSH data available.


Related in: MedlinePlus

Three percent agarose gel, Hinf I restriction fragment length polymorphism analysis of methylenetetrahydrofolate reductase 677; Lane 1-50 bp ladder; Lanes 2, 3, 4, 6 and 8 - homozygous wild type; Lane 5 and 7 -heterozygous polymorphic
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Figure 2: Three percent agarose gel, Hinf I restriction fragment length polymorphism analysis of methylenetetrahydrofolate reductase 677; Lane 1-50 bp ladder; Lanes 2, 3, 4, 6 and 8 - homozygous wild type; Lane 5 and 7 -heterozygous polymorphic

Mentions: The amplified PCR products (MTHFR) were subjected to Hinf I restriction enzyme digestion at 37°C for 1 h. The PCR products subjected to enzyme digestion was visualized on 3% agarose gel stained with ethidium bromide. Gel photography was done with bio-red gel doc system. For MTHFR 677, the PCR yielded a 315 bp product, which on digestion with Hinf I produced a 176 bp and 139 bp fragments for TT condition (homozygous polymorphic) and a 315,176 and 139 bp fragments for CT condition (heterozygous polymorphic). An undigested product length of 315 bp was retained by the wild types [Figure 2]. Results were obtained and compiled.


Methylenetetrahydrofolate reductase C677T variant in Indian children with craniosynostosis: Its role in the pathogenesis, risk of craniosynostosis.

Pandey RK, Ali A, Singh A, Gayan S, Bajpai M - Indian J Hum Genet (2014)

Three percent agarose gel, Hinf I restriction fragment length polymorphism analysis of methylenetetrahydrofolate reductase 677; Lane 1-50 bp ladder; Lanes 2, 3, 4, 6 and 8 - homozygous wild type; Lane 5 and 7 -heterozygous polymorphic
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4228567&req=5

Figure 2: Three percent agarose gel, Hinf I restriction fragment length polymorphism analysis of methylenetetrahydrofolate reductase 677; Lane 1-50 bp ladder; Lanes 2, 3, 4, 6 and 8 - homozygous wild type; Lane 5 and 7 -heterozygous polymorphic
Mentions: The amplified PCR products (MTHFR) were subjected to Hinf I restriction enzyme digestion at 37°C for 1 h. The PCR products subjected to enzyme digestion was visualized on 3% agarose gel stained with ethidium bromide. Gel photography was done with bio-red gel doc system. For MTHFR 677, the PCR yielded a 315 bp product, which on digestion with Hinf I produced a 176 bp and 139 bp fragments for TT condition (homozygous polymorphic) and a 315,176 and 139 bp fragments for CT condition (heterozygous polymorphic). An undigested product length of 315 bp was retained by the wild types [Figure 2]. Results were obtained and compiled.

Bottom Line: After successful amplification, a small aliquot (5 μl) of the MTHFR reaction mixture was treated with 1 units of Hinf I restriction enzyme (NEB).Results were obtained and compiled.When comparing the offspring of mothers statistically significant differences were found.

View Article: PubMed Central - PubMed

Affiliation: Department of Pediatric Surgery, All India Institute of Medical Sciences, New Delhi, India.

ABSTRACT

Background: 677C to T allele in the 5, 10-methylenetetrahydrofolate reductase (MTHFR) gene has been implicated in the etiology of various syndromes and nonsyndromic diseases but till date no direct studies have been reported with craniosynostosis.

Objectives: The aim was to study the family-based association of MTHFR polymorphism in different categories of craniosynostosis patients.

Materials and methods: This was a cross-sectional study in which 30 patients classified as Apert syndrome, Pfeiffr syndrome and nonsyndromic craniosynostosis patients with their family were recruited. A sample of 3 ml intravenous blood was taken from patients and from their family members (father and mother) in ethylenediaminetetraacetic acid-anticoagulated vacutainer for the purpose of the study. Genomic DNA was extracted from peripheral blood lymphocytes by phenol chloroform extraction method. Primers for MTHFR gene were designed. The polymerase chain reaction was carried out. After successful amplification, a small aliquot (5 μl) of the MTHFR reaction mixture was treated with 1 units of Hinf I restriction enzyme (NEB). Results were obtained and compiled.

Results: A total of 30 patients/participants with craniosynostosis of Indian descent and their parents formed the study group. The genotyping did not confirm an association between the MTHFR 677C to T polymorphism and between different categories of craniosynostosis. When comparing the offspring of mothers statistically significant differences were found.

Conclusion: C667T polymorphism of the MTHFR gene is unlikely to play a role in the pathogenesis of craniosynostosis though maternal MTHFR C677T polymorphism may be a genetic risk factor.

No MeSH data available.


Related in: MedlinePlus