Kenji Takakura

Mutation analysis of the growth differentiation factor-9 and -9B genes in patients with premature ovarian failure and polycystic ovary syndrome

OBJECTIVES To investigate whether growth differentiation factor (GDF)-9 or GDF-9B/bone morphogenetic protein (BMP)-15 mutation is present in Japanese women with premature ovarian failure (POF) and polycystic ovary syndrome (PCOS). DESIGN Clinical and molecular studies. SETTING Outpatient clinic in a university hospital. PATIENT(S) Fifteen women with POF, 38 women with PCOS, and 3 normal fertile controls. INTERVENTION(S) Extraction of DNA from blood samples for subsequent polymerase chain reaction (PCR). MAIN OUTCOME MEASURE(S) Amplified DNA was analyzed by denaturing gradient gel electrophoresis (DGGE) and/or by direct sequencing. RESULT(S) No missense mutation was found in any exons of the GDF-9 gene and the GDF-9B/BMP-15 gene in patients with POF and PCOS. CONCLUSION(S) The missense mutation in the GDF-9 gene or the GDF-9B/BMP-15 gene is uncommon in anovulatory Japanese women with POF and PCOS.

Follicle-stimulating hormone receptor gene mutations are rare in Japanese women with premature ovarian failure and polycystic ovary syndrome

OBJECTIVE To determine whether the known inactivating FSH receptor gene mutations are present in Japanese women with secondary amenorrhea because of premature ovarian failure (POF) and polycystic ovary syndrome (PCOS). DESIGN Clinical and molecular studies. SETTING An outpatient clinic in a university hospital. PATIENT(S) Fifteen women with idiopathic POF, 38 women with PCOS, and three normal controls. INTERVENTION(S) Extraction of DNA from blood samples for subsequent polymerase chain reaction (PCR). MAIN OUTCOME MEASURE(S) PCR fragments digested with MunI, BsmI, and HhaI were compared in patients and controls. PCR fragments were also analyzed by denaturing gradient gel electrophoresis (DGGE) and direct sequencing. RESULT(S) No inactivating mutations reported thus far in exons 6, 7, 9, and 10 of the FSH receptor gene were identified in Japanese women with POF and PCOS. DGGE analysis of PCR fragments of exon 10 also revealed no FSH receptor gene mutations in this region. CONCLUSION(S) Although we cannot exclude the presence of point mutations in other regions of the FSH receptor gene, the described FSH receptor mutations may be uncommon in Japanese patients with POF and PCOS.

Mutational analysis of the PTEN gene in women with premature ovarian failure

Recent studies in mammals have suggested that the phosphatase and tensin homolog deleted on chromosome 10 (PTEN) – phosphatidylinositol‐3, 4, 5‐trisphosphate pathway in oocytes might be related to the pathogenesis of premature ovarian failure (POF). The aim of this study was to investigate whether mutations of the PTEN gene are present in women with POF. We analyzed the coding region of the PTEN gene in 20 women with idiopathic POF and 20 normal controls. The PTEN gene was amplified by the polymerase chain reaction using genomic DNA isolated from blood samples. Amplified DNA was analyzed by denaturing gradient gel electrophoresis and direct sequencing. No causative mutation was detected in the coding regions of this gene. Although we found a point variation in exon 7 of one POF patient, this was a single nucleotide polymorphism that has already been reported.

DNA flow cytometric quantification and DNA polymorphism analysis in the case of a complete mole with a coexisting fetus.

Purpose:Our purpose was to investigate whether DNA flow cytometric quantification and DNA polymorphism analysis are useful for cytogenetic diagnosis in the case of a complete hydatidiform mole that coexists with a living fetus.Methods:Flow cytometric analysis of the nuclear DNA content and polymerase chain reaction (PCR) amplification of the minisatellite locus with the MCT118 probe were performed on the tissues (fetus, placenta and mole) obtained at the initial evacuation.Results:DNA histograms of placental, fetal, and molar tissues showed diploid peaks. PCR products demonstrated that the allele of the mole was homozygous and inherited solely from the husband and that the mole differed genetically from the fetus and the placenta.Conclusions:These results suggested that DNA flow cytometry and DNA polymorphism analysis may be useful for the cytogenetic diagnosis of a complete hydatidiform mole and a coexisting fetus.

Adenocarcinoma in situ of the cervix detected after long term follow up. A case report.

子宮頸部細胞診にて細胞異常を指摘された57ヵ月後に, 組織学的に子宮頸部上皮内腺癌に上皮内 (扁平上皮) 癌を合併して認めた1症例について報告する.症例は38歳の月経周期整なる2回経妊1回経産の女性で, 不正性器出血の既往はなく, 自覚症状は認めない.子宮頸癌集団検診にてクラスIIIaと診断されたため当科を受診した.当科における検査の結果, 子宮頸部細胞診では軽度腺細胞異型, 病理組織診では扁平上皮細胞の中等度異形成を指摘されたのみで6ヵ月ごとの外来通院にて経過観察することになった. 以後, 細胞診による腺細胞系の異常は指摘されていたが悪性を疑うような形態は認めず, 扁平上皮も異形成が消失して扁平上皮化生を認めるのみとなっていた. 4年半経過後, 細胞診にて悪性腫瘍病変を疑わせる腺細胞異型を指摘され, 精査・加療目的にて入院となる. PCR法によるHPV検査では18型と33型が陽性であった. 診断的子宮膣部円錐切除術の結果, 組織学的に上皮内腺癌と上皮内 (扁平上皮) 癌をともに認めたため, 後日単純子宮全摘術を施行した. 術後24ヵ月経過した現在, 再発徴候は認めていない.