Masaya Hirose

Targeted Disruption of the Mouse Rho-Associated Kinase 2 Gene Results in Intrauterine Growth Retardation and Fetal Death

ABSTRACT Rho-associated kinase (ROCK), including the ROCK-I and ROCK-II isoforms, is a protein kinase involved in signaling from Rho to actin cytoskeleton. However, in vivo functions of each ROCK isoform remain largely unknown. We generated mice deficient in ROCK-II by gene targeting. ROCK-II−/− embryos were found at the expected Mendelian frequency until 13.5 days postcoitum, but approximately 90% died thereafter in utero. ROCK-II−/− mice of both genders that survived were born runts, subsequently developed without gross abnormality, and were fertile. Whole-mount staining for a knocked-in lacZ reporter gene revealed that ROCK-II was highly expressed in the labyrinth layer of the placenta. Disruption of architecture and extensive thrombus formation were found in the labyrinth layer of ROCK-II−/− mice. While no obvious alteration in actin filament structures was found in the labyrinth layer of ROCK-II−/− placenta and stress fibers were formed in cultured ROCK-II−/− trophoblasts, elevated expression of plasminogen activator inhibitor 1 was found in ROCK-II−/− placenta. These results suggest that ROCK-II is essential in inhibiting blood coagulation and maintaining blood flow in the endothelium-free labyrinth layer and that loss of ROCK-II leads to thrombus formation, placental dysfunction, intrauterine growth retardation, and fetal death.

Successful intrauterine treatment with radiofrequency ablation in a case of acardiac twin pregnancy complicated with a hydropic pump twin

Twin reversed arterial perfusion sequence is a serious complication of monochorionic twin pregnancy, as the pump twin that perfuses blood to the acardiac twin may experience heart failure and fetal hydrops resulting in a poor perinatal outcome. A woman with an acardiac twin pregnancy complicated by a hydropic pump twin underwent intrauterine treatment with radiofrequency ablation (RFA) at 27 weeks of gestation. Obliteration of blood flow to the acardiac twin from the pump twin was successful. Fetal hydrops resolved by the time of delivery at 32 weeks of gestation, in spite of transient deterioration, and a good postnatal outcome was achieved for the pump twin. We found that RFA was an effective intrauterine treatment for acardiac twin pregnancy and suggest that it could be introduced in cases complicated by a hydropic pump twin. Copyright

Fibrinogen Otsu I:A γ Asn319,Asp320 deletion dysfibrinogen identified in an asymptomatic pregnant woman

Fibrinogen Otsu I:A γ Asn319,Asp320 deletion dysfibrinogen identified in an asymptomatic pregnant woman -

In utero diagnosis of an aneurysm of the vein of Galen causing hydrocephalus and heart failure.

A prenatally diagnosed aneurysm of the vein of Galen was presented in the fetus of a patient referred to our hospital at 31 weeks of gestation. Ultrasonography demonstrated polyhydramnios, cardiomegaly, dilatation of the right atrium and superior vena cava, tricuspid valve regurgitation, hydrocephalus, and a large hypoechoic mass with blood flow in the suboccipital region. Skin edema was shown thereafter. A 3262‐g male was delivered by cesarean at 35 weeks of gestation. Computed tomography imaging demonstrated a large mass in the suboccipital region, after which thrombocytopenia appeared and the neonate died at 18 days of age.

Fetal hemolytic disease due to anti-Rh17 alloimmunization

Objective: To delineate clinical features of a case of fetal hemolytic disease due to anti-Rh17, along with a review of relevant studies published in English and Japanese. Methods: We present clinical features of a –D-/-D- phenotype woman with anti-Rh17 alloimmunization during pregnancy. Relevant English literature in the MEDLINE database was reviewed, while Japanese studies were searched in the Japana Centra Revuo Medicina database. Results: A Japanese –D-/-D- woman with anti-Rh17 (Hro) was treated during pregnancy. Serial ultrasonography, antibody titers, amniocenteses, and cordocenteses were conducted for perinatal management. Amniocentesis results demonstrated a high delta optical density level of 450 in the amniotic fluid, while cordocentesis revealed alloimmunization between the mother and the fetus as well as fetal hemolytic anemia. Blood flow velocity in the middle cerebral artery indicated a rapid development of fetal anemia. The newborn demonstrated severe anemia and hyperbilirubinemia, which were successfully treated with exchange transfusions. Two cases of prenatally diagnosed fetal hemolytic disease due to anti-Rh17 were found published in English and 5 in Japanese. Conclusion: A –D-/-D- phenotype patient with anti-Rh17 was successfully managed during pregnancy and a good outcome for the neonate was achieved. Our results and a review of related literature led to the following suggestions. The first pregnancy in a –D-/-D- woman may be affected, an anamnestic immune response can easily occur during pregnancy, the level of anti-Rh17 titer is indicative of the degree of fetal hemolysis, and appropriate intrauterine intervention is warranted for achievement of a good outcome.

Protein S gene mutation in a young woman with type III protein S deficiency and venous thrombosis during pregnancy.

AbstractBackground: We attempted to identify a gene defect in a young woman with type III protein S deficiency and venous thrombosis during pregnancy. Methods: Measurements of total and free PS antigen levels in plasma were carried out using an enzyme-linked immunosorbent assay. Plasma PS cofactor activity was determined by a clotting assay using activated factor V as the substrate. Genomic DNA prepared from peripheral blood was amplified by polymerase chain reaction (PCR) with PROS1-specific oligonucleotide primers. PCR products were sequenced on both strands using specific oligonucleotide primers. Results: Plasma PS cofactor activity was undetectable in every measurement at 36 weeks of gestation, as well as at 2 weeks and 4 months after delivery. Plasma total PS antigen levels were 70% and 67% at 2 weeks and 4 months after delivery, respectively. Free PS antigen level was 24% at 4 months after delivery. Of all exons analyzed, codon 295 of GGC in exon 10 was substituted for AGC. This missense mutation predicted an amino acid change of glycine to serine. Conclusions: Measurements of total and free PS antigen levels along with PS activity indicated that this was a case of type III PS deficiency. DNA analysis identified a heterozygous missense mutation of codon 295 in the PS gene, substituting glycine for serine.

Current status of non‐invasive prenatal testing in Japan

The purpose of this study was to report the 3‐year experience of a nationwide demonstration project to introduce non‐invasive prenatal testing (NIPT) of maternal plasma for aneuploidy, and review the current status of NIPT in Japan.

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.