Diagnosis of Hemophilia B Carriers Using Two Novel Dinucleotide Polymorphisms and Hha I RFLP of the Factor IX Gene in Japanese Subjects

Toyozumi, Hisato; Kojima, Tetsuhito; Matsushita, Tadashi; Hamaguchi, Motohiro; Tanimoto, Mitsune; Saito, Hidehiko

doi:10.1055/s-0038-1649870

Cited by 18 publications

(15 citation statements)

References 4 publications

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“…After obtaining informed consent from all patients, blood samples from the patients, family members, and healthy volunteers were collected in a 1/ 10 volume of 3.13% sodium citrate. Plasma was separated by centrifugation at Â 2000g for 20 min, and genomic DNA was isolated from peripheral blood leukocytes as previously described [7].…”

Section: Patients and Sample Preparationsmentioning

confidence: 99%

Molecular basis of antithrombin deficiency in four Japanese patients with antithrombin gene abnormalities including two novel mutations

et al. 2007

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We analyzed the antithrombin (AT) gene in four unrelated Japanese patients with an AT deficiency, and individually identified four distinct mutations in the heterozygous state. There were two novel mutations, 2417delT leading to a frameshift with a premature termination at amino acid -3 (FS-3Stop) and C2640T resulting in a missense mutation (Ala59Val). Previously reported mutations, T5342C (Ser116Pro) and T72C (Met-32Thr), were also found in the other two patients. To understand the molecular basis responsible for the AT deficiency in these patients, in vitro expression experiments were performed using HEK293 cells transfected with either wild type or respective mutant AT expression vector. We found that -3Stop-AT and -32Thr-AT were not secreted into the culture media, whereas 116Pro-AT and 59Val-AT were secreted normally. We further studied the heparin cofactor activity and the binding to heparin of each recombinant AT molecule. Ser116Pro mutation significantly impaired the binding affinity to heparin resulting in a reduced heparin cofactor activity. In contrast, we found that Ala59Val mutant AT unexpectedly showed a normal affinity to heparin, but severely impaired the heparin cofactor activity. Our findings suggested that FS-3Stop and Met-32Thr mutations are responsible for type I AT deficiency, whereas Ser116Pro and Ala59Val mutations contribute to type II AT deficiency, confirming that there were diverse molecular mechanisms of AT deficiency depend upon discrete AT gene abnormalities as reported previously. Am. J. Hematol. 82:702-705, 2007. V

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Section: Patients and Sample Preparationsmentioning

confidence: 99%

Molecular basis of antithrombin deficiency in four Japanese patients with antithrombin gene abnormalities including two novel mutations

et al. 2007

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“…It was also mentioned that although the study of restriction fragment length polymorphisms is the preferred method in familial haemophilia, it is less useful in isolated cases [14]. DNA polymorphism studies also revealed that some single polymorphic sites can assess the carrier status up to approximately 30% whereas three or more probes can give the results up to 90% [15][16][17][18][19]. In another study, racial variations were also seen, i.e.…”

Section: Resultsmentioning

confidence: 99%

Assessment of the carrier status by pedigree analysis in some families from India

Singh

Kaur

2002

Haemophilia

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Pedigree analysis is an important tool to assess the carrier status of the females in the families of haemophiliac patients. A study was conducted on 85 families, of which 75 were of haemophilia type A and 10 of haemophilia type B. The probability values of the females being carriers were calculated by the Bayesian method. The results revealed that 45 mothers were genetic obligate carriers and the remaining 40 probable, hence the transmission was of familial nature in 45 families, whereas the remaining 40 were categorized as isolated cases. The probability values of 425 females were calculated and a wide range of probability values ranging from 0.0295 to 1 was observed.

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“…To analyse the X chromosome haplotype of the family members, we used polymorphic markers, such as monoamine oxidase A gene ( MAOA , Xp11.23) promoter [18], androgen receptor gene ( AR , Xq11–12) [19], phosphoglycerate kinase 1 gene ( PGK1 , Xq13.3) [16], glutamate receptor ionotropic AMPA 3 gene ( GRIA3 , Xq25–26) [17], FIX gene ( F9 ‐ 793, Xq27.1–2) [20] and Fragile X mental retardation gene 1 ( FMR1 , Xq27.3) [16]. We also analysed short tandem repeat [13] polymorphisms, such as reference SNP IDs: rs3223331 (Xq22), rs3220745 (Xq24) and rs3220178 (Xq26).…”

Section: Patients Materials and Methodsmentioning

confidence: 99%

Skewed X chromosome inactivation in fraternal female twins results in moderately severe and mild haemophilia B

et al. 2008

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Female carriers of haemophilia B are usually asymptomatic; however, the disease resulting from different pathophysiological mechanisms has rarely been documented in females. In this study, we investigated the mechanisms responsible for haemophilia B in fraternal female twins. We sequenced the factor IX gene (F9) of the propositus, her father, a severe haemophilia B patient and the other family members. X chromosome inactivation was assessed by the methylation-sensitive HpaII-PCR assay using X-linked polymorphisms in human phosphoglycerate kinase 1 gene (PGK1) and glutamate receptor ionotropic AMPA 3 gene (GRIA3). The twins were found to be heterozygotes with a nonsense mutation (p.Arg384X) inherited from their father. The propositus, more severely affected twin, exhibited a significantly higher percentage of inactivation in the maternally derived X chromosome carrying a normal F9. The other twin also showed a skewed maternal X inactivation, resulting in a patient with mild haemophilia B. Thus, the degree of skewing of maternal X inactivation is closely correlated with the coagulation parameters and the clinical phenotypes of the twins. Furthermore, we identified a crossing-over in the Xq25-26 region of the maternal X chromosome of the more severely affected twin. This crossing-over was absent in the other twin, consistent with their fraternal state. Differently skewed X inactivation in the fraternal female twins might cause moderately severe and mild haemophilia B phenotypes, respectively.

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Diagnosis of Hemophilia B Carriers Using Two Novel Dinucleotide Polymorphisms and Hha I RFLP of the Factor IX Gene in Japanese Subjects

Cited by 18 publications

References 4 publications

Molecular basis of antithrombin deficiency in four Japanese patients with antithrombin gene abnormalities including two novel mutations

Molecular basis of antithrombin deficiency in four Japanese patients with antithrombin gene abnormalities including two novel mutations

Assessment of the carrier status by pedigree analysis in some families from India

Skewed X chromosome inactivation in fraternal female twins results in moderately severe and mild haemophilia B

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