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Purpose Although non‐invasive prenatal testing (NIPT) based on cell‐free DNA (cfDNA) in maternal plasma has been prevailing worldwide, low levels of fetal DNA fraction may lead to false‐negative results. Since fetal cells in maternal blood provide a pure source of fetal genomic DNA, we aimed to establish a workflow to isolate and sequence fetal nucleated red blood cells (fNRBCs) individually as a target for NIPT. Methods Using male‐bearing pregnancy cases, we isolated fNRBCs individually from maternal blood by FACS, and obtained their genomic sequence data through PCR screening with a Y‐chromosome marker and whole‐genome amplification (WGA)‐based whole‐genome sequencing. Results The PCR and WGA efficiencies of fNRBC candidates were consistently lower than those of control cells. Sequencing data analyses revealed that although the majority of the fNRBC candidates were confirmed to be of fetal origin, many of the WGA‐based genomic libraries from fNRBCs were considered to have been amplified from a portion of genomic DNA. Conclusions We established a workflow to isolate and sequence fNRBCs individually. However, our results demonstrated that, to make cell‐based NIPT targeting fNRBCs feasible, cell isolation procedures need to be further refined such that the nuclei of fNRBCs are kept intact.

The incidence of chromosomal abnormalities in twin pregnancies is not well-studied. In this retrospective study, we investigated the frequency of chromosomal abnormalities in twin pregnancies and compared the incidence of chromosomal abnormalities in dichorionic diamniotic (DD) and monochorionic diamniotic (MD) twins. We used data from 57 clinical facilities across Japan. Twin pregnancies of more than 12 weeks of gestation managed between January 2016 and December 2018 were included in the study. A total of 2899 and 1908 cases of DD and MD twins, respectively, were reported, and the incidence of chromosomal abnormalities in one or both fetuses was 0.9% (25/2899) and 0.2% (4/1908) in each group (p = 0.004). In this study, the most common chromosomal abnormality was trisomy 21 (51.7% [15/29]), followed by trisomy 18 (13.8% [4/29]) and trisomy 13 (6.9% [2/29]). The incidence of trisomy 21 in MD twins was lower than that in DD twins (0.05% vs. 0.5%, p = 0.007). Trisomy 21 was less common in MD twins, even when compared with the expected incidence in singletons (0.05% vs. 0.3%, RR 0.15 [95% CI 0.04–0.68]). The risk of chromosomal abnormality decreases in twin pregnancies, especially in MD twins.

Aim We aimed to evaluate the sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of noninvasive prenatal testing (NIPT) in high-risk pregnant women. Methods Pregnant women who underwent GeneTech NIPT, the most commonly used NIPT in Japan, between January 2015 and March 2019, at Japan NIPT Consortium medical sites were recruited for this study. The exclusion criteria were as follows: pregnant women with missing survey items, multiple pregnancy/vanishing twins, chromosomal abnormalities in the fetus other than the NIPT target disease, and nonreportable NIPT results. Sensitivity and specificity were calculated from the obtained data, and maternal age-specific PPV and NPV were estimated. Results Of the 45 504 cases, 44 263 cases fulfilling the study criteria were included. The mean maternal age and gestational weeks at the time of procedure were 38.5 years and 13.1 weeks, respectively. Sensitivities were 99.78% (95% confidence interval [95% CI]: 98.78-99.96), 99.12% (95% CI: 96.83-99.76), and 100% (95% CI: 88.30-100) for trisomies 21, 18, and 13, respectively. Specificities were more than 99.9% for trisomies 21, 18, and 13, respectively. Maternal age-specific PPVs were more than 93%, 77%, and 43% at the age of 35 years for trisomies 21, 18, and 13, respectively. Conclusion The GeneTech NIPT data showed high sensitivity and specificity in the detection of fetal trisomies 21, 18, and 13 in high-risk pregnant women, and maternal age-specific PPVs were obtained. These results could provide more accurate and improved information regarding NIPT for genetic counseling in Japan.

Aim 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. Methods Tests were conducted to detect aneuploidy in high-risk pregnant women, and adequate genetic counseling was provided. The clinical data, test results, and pregnancy outcomes were recorded. We discuss the problems of NIPT on the basis of published reports and meta-analyses. Results From April 2013 to March 2016, 30 613 tests were conducted at 55 medical sites participating in a multicenter clinical study. Among the 30 613 women tested, 554 were positive (1.81%) and 30 021 were negative (98.1%) for aneuploidy. Of the 289, 128, and 44 women who tested positive for trisomies 21, 18, and 13, respectively, and underwent definitive testing, 279 (96.5%), 106 (82.8%), and 28 (63.6%) were determined to have a true-positive result. For the 13 481 women with negative result and whose progress could be traced, two had a false-negative result (0.02%). The tests were performed on the condition that a standard level of genetic counseling be provided at hospitals. Conclusion Here, we report on the 3-year nationwide experience with NIPT in Japan. It is important to establish a genetic counseling system to enable women to make informed decisions regarding prenatal testing. Moreover, a welfare system is warranted to support women who decide to give birth to and raise children with chromosomal diseases.

Aim The aim of this study was to investigate the practicability and efficiency of lectin-based isolation of fetal erythroblasts for clinical use in non-invasive prenatal testing. Methods Peripheral blood samples were collected from 39 pregnant women. Leukocytes were removed with an anti-CD45 antibody after density gradient centrifugation. After blood cells were attached to slides by binding to a galactose-specific lectin and galactose-bound vinyl polymer, the slides were stained with May–Grunwald–Giemsa stain and cells were classified by automated image analysis based on their size and the nuclear area/cytoplasmic area ratio. In 14 samples from the women with male fetuses, fetal origin of the isolated erythroblasts was confirmed by detecting the Y chromosome using fluorescence in situ hybridization. In eight samples, single erythroblasts were collected by the laser capture microdissection technique for amplification of the sex-determining region Y gene to confirm fetal origin. Results Panning with an anti-CD45 antibody achieved stable removal of leukocytes without aggregation. In all samples, erythroblasts were successfully identified by automated image analysis (18–6000/10 mL of blood). The number of slides required to examine 10 mL of blood ranged from one to six, which was reasonable for clinical use. The Y chromosome was detected in 7.5–43.6% of erythroblasts by fluorescence in situ hybridization, and the sex-determining region Y gene was amplified in seven of eight samples. Conclusion The combination of lectin-based erythroblast isolation and automated image analysis is a practical and efficient method for isolating fetal erythroblasts as a source of fetal genomes.

The purpose of this noninvasive prenatal testing (NIPT) study was to compare the fetal fraction of singleton gestations by gestational age, maternal characteristics and chromosome-specific aneuploidies as indicated by z-scores. This study was a multicenter prospective cohort study. Test data were collected from women who underwent NIPT by the massively parallel sequencing method. We used sequencing-based fetal fraction calculations in which we estimated fetal DNA fraction by simply counting the number of reads aligned within specific autosomal regions and applying a weighting scheme derived from a multivariate model. Relationships between fetal fractions and gestational age, maternal weight and height, and z-scores for chromosomes 21, 18 and 13 were assessed. A total of 7740 pregnant women enrolled in the study, of which 6993 met the study criteria. As expected, fetal fraction was inversely correlated with maternal weight (P

OBJECTIVE To evaluate the reasons for nonreportable cell-free DNA (cfDNA) results in noninvasive prenatal testing (NIPT), we retrospectively studied maternal characteristics and other details associated with the results. METHODS A multicenter retrospective cohort study in pregnant women undergoing NIPT by massively parallel sequencing (MPS) with failed cfDNA tests was performed between April 2013 and March 2017. The women's data and MPS results were analyzed in terms of maternal characteristics, test performance, fetal fraction (FF), z scores, anticoagulation therapy, and other details of the nonreportable cases. RESULTS Overall, 110 (0.32%) of 34 626 pregnant women had nonreportable cfDNA test results after an initial blood sampling; 22 (20.0%) cases had a low FF (

Objective The purpose of this study is to compare the fetal fractions during non-invasive prenatal testing (NIPT) in singleton pregnancies according to gestational age and maternal characteristics to evaluate the utility of this parameter for the prediction of pregnancy complications including gestational diabetes mellitus (GDM) and hypertensive disorders of pregnancy (HDP). Study design This study was a multicenter prospective cohort study. The present data were collected from women whose NIPT results were negative. The relationships between the fetal fractions and the gestational age, maternal weight and height, and incidences of miscarriage, preterm delivery, and pregnancy complications including GDM, HDP and placental abruption were assessed. Results A total of 5582 pregnant women with verified NIPT negative results were registered in the study. The demographic characteristics of the study populations were statistically analyzed, and the women with HDP tended to have a low fetal fraction in samples taken during early gestation. The area under the curve (AUC) in a receiver operating characteristic curve (ROC) analysis was 0.608 for women with HDP. Conclusion A low fetal fraction on NIPT might be correlated with future HDP. However, predicting HDP during early pregnancy in women with a low fetal fraction might be difficult.

Hypertensive disorders of pregnancy are known to be a risk factor for future cardiovascular diseases. In contrast, there is a paucity of data on the not so distant future prognosis of hypertensive disorders of pregnancy. In the present study, we evaluated the incidence of the diseases causing cardiovascular problems (hypertension, diabetes mellitus, dyslipidemia and metabolic syndrome) 5 years after delivery in Japanese women with hypertensive disorders of pregnancy. We performed a double-cohort study and compared medical conditions between women with and without a history of hypertensive disorders of pregnancy. A total of 1513 women who participated in the cohort study were invited to undergo a medical checkup 5 years after the index delivery, of whom 829 responded. After excluding pregnant and lactating women at the time of examination, 25 women with hypertensive disorders of pregnancy and 746 control subjects were analyzed. The incidence of hypertension was significantly higher among women with hypertensive disorders of pregnancy than women who were normotensive during pregnancy (24.0 vs. 2.5%, P

β-thalassemia is one of the most common genetic disorders worldwide. Concerted efforts are being made to prevent the disease, as the medical and economic burden of thalassemia represents a major public health problem. The molecular diagnosis of the β-globin mutations that cause the disease currently involves a combination of classic methodologies. A microarray-based assay for parallel one-shot detection of mutations has been developed, but the assay remains too expensive for routine application. We developed a cost-effective plastic fiber-based DNA chip for the fast and reliable detection of 25 types of β-thalassemia mutations. Assay conditions were established and genotyping was successfully performed on a genomic sample from a β-thalassemia patient. Our data show that this β-thalassemia genotyping chip is an advantageous platform for mass genotyping because of its low cost, rapid results, and reliability.

BACKGROUND Selection of hematopoietic stem cells can be used to prevent graft-versus-host disease (GVHD) after allograft transplantation. The purpose of the study was to examine a novel cell separation system comprising a galactose-bound vinyl polymer (Gal-VP) and soybean agglutinin (SBA), a galactose-specific lectin. STUDY DESIGN AND METHODS A vinyl polymer (VP) containing alpha-1,6- and beta-1,4-linked galactose terminals was used to facilitate cell separation. A VP containing an alpha-1,4-linked glucose terminal (alpha-1,4-Glu-VP) was also synthesized as a control for alpha-1,6- and beta-1,4-Gal-VP. Peripheral blood samples were collected from healthy volunteers and umbilical cord blood cells were collected after normal labor. RESULTS The sugar-VP was adsorbed on the surface of various materials. In the presence of SBA, T lymphocytes bound to beta-1,4-Gal-VP-coated microbeads, but not to alpha-1,4-Glu-VP-coated microbeads. When peripheral or cord blood cells were cultured on alpha-1,6-Gal-VP-coated plates, most red blood cells, lymphocytes, granulocytes, and monocytes adhered to the plate in the presence of 300 mg per mL SBA, whereas few CD34+ cells attached, even with 800 mg per mL SBA. CONCLUSION SBA binds selectively to blood cells by recognizing cell-surface sugars, which are dependent on the extent of cellular differentiation. Therefore, the combination of alpha-1,6-Gal-VP and SBA might be useful for separation of blood cells according to their stage of differentiation and lineage.

Anterior urethral valves are a rare congenital anomaly associated with distal urethral obstruction, which can result in a poor prognosis. We report on the endoscopic creation of a fetal urethrotomy for obstructive uropathy resulting from anterior urethral valves. A 33-year-old woman was evaluated at 17 weeks gestation due to fetal megacystis. The diagnosis of anterior urethral valves was confirmed by the characteristic sonographic feature of a dilated membranous penile urethra. Oligohydramnios with normal-appearing kidneys and favorable urinary electrolytes led to fetal intervention. Ablation on the ventral site of the fetal penis for a cutaneous urethrotomy was performed using a YAG laser under a 1-mm fetoscope at 19 weeks gestation. Urine was drained from the incision and the dilated penis and the distended bladder shrunk with an increase in amniotic fluid. However, the fetus died unexpectedly on postoperative day 3, and chorioamnionitis was suspected as the etiology. While the outcome was unfavorable, our preliminary experience shows that fetal urethrotomy for obstructive uropathy can be achieved in utero using an endoscopic laser approach. Further experience will be required to evaluate the therapeutic value of this new procedure in the management of fetal anterior urethral valves.

The purpose of this study is to summarize the results from a survey on awareness of genetic counseling for pregnant women who wish to receive non-invasive prenatal testing (NIPT) in Japan. As a component of a clinical study by the Japan NIPT Consortium, genetic counseling was conducted for women who wished to receive NIPT, and a questionnaire concerning both NIPT and genetic counseling was given twice: once after pre-test counseling and again when test results were reported. The responses of 7292 women were analyzed. They expressed high satisfaction with the genetic counseling system of the NIPT Consortium (94%). The number of respondents who indicated that genetic counseling is necessary for NIPT increased over time. Furthermore, they highly valued genetic counseling provided by skilled clinicians, such as clinical geneticists or genetic counselors. The vast majority (90%) responded that there was sufficient opportunity to consider the test ahead of time. Meanwhile, women who received positive test results had a poor opinion and expressed a low-degree satisfaction. We confirmed that the pre-test genetic counseling that we conducted creates an opportunity for pregnant women to sufficiently consider prenatal testing, promotes its understanding and has possibilities to effectively facilitate informed decision making after adequate consideration. A more careful and thorough approach is considered to be necessary for women who received positive test results.

The aim of this study was to investigate the practicability and efficiency of lectin-based isolation of fetal erythroblasts for clinical use in non-invasive prenatal testing.Peripheral blood samples were collected from 39 pregnant women. Leukocytes were removed with an anti-CD45 antibody after density gradient centrifugation. After blood cells were attached to slides by binding to a galactose-specific lectin and galactose-bound vinyl polymer, the slides were stained with May-Grünwald-Giemsa stain and cells were classified by automated image analysis based on their size and the nuclear area/cytoplasmic area ratio. In 14 samples from the women with male fetuses, fetal origin of the isolated erythroblasts was confirmed by detecting the Y chromosome using fluorescence in situ hybridization. In eight samples, single erythroblasts were collected by the laser capture microdissection technique for amplification of the sex-determining region Y gene to confirm fetal origin.Panning with an anti-CD45 antibody achieved stable removal of leukocytes without aggregation. In all samples, erythroblasts were successfully identified by automated image analysis (18-6000/10 mL of blood). The number of slides required to examine 10 mL of blood ranged from one to six, which was reasonable for clinical use. The Y chromosome was detected in 7.5-43.6% of erythroblasts by fluorescence in situ hybridization, and the sex-determining region Y gene was amplified in seven of eight samples.The combination of lectin-based erythroblast isolation and automated image analysis is a practical and efficient method for isolating fetal erythroblasts as a source of fetal genomes.

Aim: We performed in vivo and in vitro adhesion experiments on the use of photocrosslinkable chitosan (PC) for healing of the myometrium and the amniotic membrane to test its usefulness in the perinatal field. Methods: To determine the adhesion conditions of PC in the healing of the myometrium and amniotic membrane, we applied two concentrations (30 and 50 mg/mL) of PC to the postpartum amniotic membrane, and radiated it with ultraviolet (UV) light for 10, 20, 30 or 60 s. Next, we created incisions of three sizes (pinhole, 5 mm and 10 mm), and performed PC adhesion followed by pressure application experiments. Subsequently, we laparotomized 12-day-pregnant rats under anesthesia, created a 5-mm incision, applied PC and closed the abdomen. We removed the uterus at 20 days of pregnancy, and histologically examined the site of PC application. Results: The optimal adhesion conditions of PC were 50 mg/mL, and the optimal time of UV irradiation was 30 s or longer. In pressure application experiments, the bonding strength of PC was so high that even 10-mm incisions withstood a mean pressure of 92 mmH2O. The bonding strength of PC was significantly higher than that of fibrin glue that was tested in parallel. Histological examination of the rat myometrium showed a mild foreign body reaction with no inflammatory findings, confirming the usefulness of PC. Conclusions: PC is a wound-healing drug, an improvement on the naturally occurring polysaccharide chitosan, and is safer and has a higher bonding strength than biological adhesives including fibrin, making it promising for fetal surgical and perinatal applications.

Currently, the standard method of diagnosis of twin reversed arterial perfusion (TRAP) sequence is ultrasound imaging. The use of MRI for flow visualization may be a useful adjunct to US imaging for assessing the presence of retrograde blood flow in the acardiac fetus and/or umbilical artery. The technical challenge in fetal MRI flow imaging, however, is that fetal electrocardiogram (ECG) monitoring required for flow imaging is currently unavailable in the MRI scanner. A non-gated MRI flow imaging technique that requires no ECG monitoring was developed using the t-test to detect blood flow in 20 slices of phase-contrast MRI images randomly scanned at the same location over multiple cardiac cycles. A feasibility study was performed in a 24-week acardiac twin that showed no umbilical flow sonographically. Non-gated MRI flow images clearly indicated the presence of blood flow in the umbilical artery to the acardiac twin; however, there was no blood flow beyond the abdomen. This study leads us to conjecture that non-gated MRI flow imaging is sensitive in detecting low-range blood flow velocity and can be an adjunct to Doppler US imaging.

Non-invasive prenatal diagnostic methods posing no danger to the embryo have been desired for many years in the field of prenatal medicine. We are in the process of improving the lectin method that we developed for recovering fetus-derived nucleated red blood cells (NRBC) in the maternal peripheral blood. We previously used Ficoll density-gradient centrifugation for preliminary concentration in the lectin method. In the present study, we developed a molecular filter method, compared it with the Ficoll method, and tested its applicability to prenatal diagnosis. We tested the usefulness of a high molecular filter method for preliminary concentration. First, in a basic study, we prepared three kinds of non-woven cloth (NWC1-3) and a multi-porous filter (MP) to determine the optimal filter. Next, we compared the recovery rates of the Ficoll and filter methods as preliminary concentration methods in 34 normal pregnant women. Then, to examine whether the recovered NRBC were derived from the fetus, we attempted prenatal diagnosis by the fluorescence in situ hybridization (FISH) technique in 12 women pregnant with a male fetus (determined later by ultrasound) at between 8 and 16 weeks of gestation. Among the four devices used in the basic study of the high molecular filter method, NWC-2 had the best recovery rate. Therefore, we compared the numbers of NRBC recovered by the lectin method after preliminary concentration with NWC-2 or by Ficoll centrifugation, and found that the mean recovery rate of NWC-2 was 4.2 +/- 5.0 times as high as that of the Ficoll method, indicating that the NWC-2 filter method is superior as a preliminary concentration method. Next, FISH analysis of the 12 pregnant women with a male fetus for the Y chromosome showed that 19.5 +/- 12.8 NRBC were recovered, in 12.7 +/- 8.1 (63.6%) of which a Y signal was confirmed, suggesting the NWC-2 filter method can be applied to prenatal diagnosis. We consider the filter-lectin method to be a superior method for isolation and recovery of NRBC in the maternal blood which can be applied to prenatal diagnosis.

To isolate fetal cells from maternal blood, we developed a new method based on galactose-bearing conjugation. Nucleated red blood cells (NRBCs), which highly express galactose on their surface, were selectively attached to a substrate coated with a galactose-containing polymer via soybean agglutinin (SBA), a galactose-specific lectin. Cord blood samples were used to evaluate enrichment efficacy of NRBCs by this method. Blood samples were obtained from 131 pregnant women between 6 and 27 gestational weeks. After preliminary condensation of fetal cells by Ficoll gradient centrifugation, NRBCs were enriched using galactose-positive selection by adjusting SBA concentration. We isolated one to several hundred NRBCs (mean+/-SD, 7.8+/-8.5) in 2.3 ml of peripheral blood samples from 96% of pregnant women. The isolated NRBCs were analyzed by a Y-chromosome FISH probe in eight cases carrying male fetuses. Y-signals were detected in all eight cases and more than half of the NRBCs were off fetal origin. The study demonstrates that our new method using galactose-specific lectin provides effective enrichment of fetal NRBCs allowing non-invasive prenatal diagnosis.

Objective:To evaluate vascular endothelial-cell functioning in placentas with gestosis, using the monoclonal antibody to cadherin 5. Methods:The extra-cellular moiety of cadherin 5 was transfected into L-cells to enable us to examine their cell-adhesion activity. The expression of cadherin 5 was evaluated in human umbilical-vein endothelial cells and in placentas with gestosis by immunostaining using the anti-cadherin 5 antibody. A microspectrophotometric study also was conducted of the placentas with gestosis. Results:We determined the total base sequence for cadherin 5 and found that it is homologus with a known cadherin but is a new, unique clone. Cadherin 5 has cell-adhesion activity and is expressed in endothelial cells at the cell-adhesion surface. The expression of cadherin 5 in endothelial cells took place in the placentas with gestosis, but to a lesser extent than in normal placental endothelial cells. Conclusions:The reduced expression of cadherin 5 in placentas with gestosis suggests that endothelial cell functioning is impaired in placentas with gestosis. Cadherin 5 in endothelial cells might influence placental functions and fetal development.

Objective To assess the efficiency and reliability of the separation of fetal nucleated red blood cells (NRBCs) using the galactose-specific lectin method, we counted the number of NRBCs in the blood of pregnant women at various gestational ages, as well as after amniocentesis and termination. Method Peripheral blood samples were obtained from (1) 22 singleton pregnant women (between 9 and 34 weeks of gestation) and from 23 women who underwent termination (between 6 and 19 weeks of gestation). To determine whether amniocentesis influences numbers of NRBCs, five samples were obtained (2) before and after the procedure. NRBC enrichment was initially performed using density gradients and subsequently using galactose-specific lectin. The cells were then stained with May-Gruenwald Giemsa (MGG) and counted under a light microscope. Results NRBCs were found in all samples, ranging from 1 to 82 (median = 12.5 cells/sample). The multiples of the median (MoM) conversion of the number of cells revealed a raise of 1.66-fold (0.12–6.64) in post-termination samples compared with the control value of 1.00 MoM (0.11–6.92; p = 0.036). The postamniocentesis increase was, instead, 1.11-fold (0.17–4.02), which did not reach statistical significance. Conclusion All blood samples tested contained NRBCs. Samples obtained after termination yielded more cells than those obtained from women whose pregnancies were going on normally. The number of NRBCs in post-termination samples after MoM conversion differed significantly from those in controls. Although separation of NRBCs was not feasible due to extremely low numbers, our results indicated that NRBCs are detectable in all blood samples from normal pregnant women. Copyright © 2006 John Wiley & Sons, Ltd.

We studied the preventive effect against allergies in infants who and whose mothers consumed hypoallergenic formulas until 6 months after birth. Mother and infant pairs were divided into three groups, and the infants were monitored for the development of allergies for the first 2 years. In the MD group (n = 102; n = number of infants), the mothers were given a hypoallergenic formula for mothers (MOM HA), which contained hydrolyzed whey protein as the only protein source, as a substitution for cow's milk during late pregnancy and lactation. In the CD group (n = 127), the mothers were given cow's milk during the corresponding period. All infants in the MD and CD groups were exclusively breast-fed or mixed-fed with breast milk and hypoallergenic infant formula (NAN HA), which contains the same hydrolyzed protein as MOM HA. In the AF group (n = 54), the mothers consumed MOM HA and their infants were mixed-fed with breast milk and a cow's milk-based adopted infant formula during the corresponding period. In the MD group, no infants were positive to cow's milk-specific immunoglobulin E (RAST) at 4 months of age, in contrast to 6% and 3% of infants in the CD and AF groups, respectively. The infants in the MD group showed low incidence of various allergies, especially of eczema, as compared to the CD and AF groups. These results suggest that consumption of cow's milk by mothers and cow's milk-based formula feeding to infants elevate the risk of allergies in infants, and that consumption of hypoallergenic formula for pregnant and lactating women and for infants could be helpful in preventing allergy development in infants.

Food antigens transferred into breast milk sometimes cause an allergic reaction in exclusively breast-fed infants. This study will show whether the intake of a whey hydrolysate formula for lactating women (MOM HA) can reduce the appearance of food antigens in breast milk. Lactating women in the MOM group (n = 12) consumed MOM HA as a substitute for cow's milk and those in the COW group (n = 13) consumed cow's milk for more than 4 months. After the ingestion of 200 mL of MOM HA and cow's milk by the women in the MOM and COW groups, respectively, the first breast milk samples were obtained and beta-lactoglobulin was measured using enzyme-linked immunosorbent assay. The number of subjects with detectable beta-lactoglobulin (> 0.1 ng/mL) in the MOM group was two (17%), which was significantly less than that in the COW group (11 subjects, 85%, p < 0.01). The level of beta-lactoglobulin was also lower in the MOM group than the COW group (p < 0.01). Subsequently, the women in the MOM group consumed cow's milk and those in the COW group consumed MOM HA for one week; then a second sampling was performed. beta-Lactoglobulin was detected in three (25%) and 8 subjects (62%) in the MOM and COW groups, respectively. The level of beta-lactoglobulin was still lower in the MOM group (p < 0.05). The consumption of whey hydrolysate formula by lactating women over a considerable time reduces the transfer of beta-lactoglobulin into their breast milk, and the low level can be maintained even after inadvertent ingestion of cow's milk.

Background: Reports of pregnancy in acromegalic women are uncommon, numbering less than 100, in which a case of acromegaly first diagnosed in pregnancy is rare. Case: A 40-year-old woman in 19-week gestation was presented with a 10-week history of photophobia and visual disturbance. Elevated growth hormone (GH) levels in the serum, an inadequate suppression of GH secretion with glucose administration and a mass image in the pituitary fossa confirmed the diagnosis of acromegaly. We initiated dopamine agonist treatment, bromocriptine at a weekly dose of 1 mg at 27-week gestation. Ten weeks after the start of the bromocriptine treatment, the GH levels gradually decreased to a normal range, and visual symptoms were improved. Enlargement of the tumor was not observed throughout the pregnancy and the patient delivered a healthy infant following an eventful pregnancy. At 3 weeks after delivery, she underwent a transsphenoidal resection of a pituitary adenoma. Conclusion: Pregnancy may lead to pituitary tumor growth and increase the risk of developing loss of vision. It is important to take pituitary tumors into consideration when encountering gravida with visual disturbance.

[Background] Hypertensive disease in pregnancy (HDP) is associated with a risk of subsequent hypertension. However the influence of normotensive blood pressure (BP) during pregnancy on future BP is not clear. [Purpose] To investigate the association between BP measurements at their lowest levels in pregnancy, 20 weeks’ gestation (20 wks BP), and the risk of hypertension both ①during pregnancy and ②5 years after delivery. [Methods] A total of 1542 women who delivered singletons at the National Center for Child Health and Development and Showa University Hospital were recruited between 2003 and 2007. They were invited to undergo a physical examination 5 years after delivery. Following exclusions (see below), 772 women completed the analysis. The influence of 20 wks BP on HDP and 5-year hypertension was assessed by multiple logistic regression analysis. The odds ratio for HDP was adjusted by age at delivery, pre-pregnancy BMI, familial history of hypertension (FH), previous history of HDP and parity (Model 1) and that for subsequent hypertension was adjusted by existence of HDP in index pregnancy, age, BMI, FH and renal disease (Model 2). [Criteria] HDP was defined as sBP ≥ 140 mm Hg or dBP ≥ 90 mm Hg from 20 weeks’ gestation which remits by 12 weeks postpartum w/o proteinuria. Hypertension at physical examination was defined as average sBP ≥ 140 mm Hg or average dBP ≥ 90 mm Hg or treatment with antihypertensive agents. [Exclusion criteria] Chronic hypertension / Women who were pregnant and nursing when the physical examination was done [Results] There were 26 HDP cases and 25 cases of 5-year hypertension (HDP: 6, normotensive control: 19). ① The odds ratio (95%CI) for HDP (adjusted by Model 1) is sBP:1.11 (for every 1 mmHg rise) (1.05~1.17), dBP:1.15(1.06~1.24) and BP category defined by the American Heart Association : 5.82 (for every 1 category increase)(2.15~15.76). A positive correlation between 20 wks BP and HDP risk was observed. ② The odds ratio for subsequent hypertension (adjusted by Model 2) is sBP: 1.06 (1.01-1.11), dBP:1.15(1.06-1.24) and BP category: 4.50(1.64-12.33). 20 wks BP is associated with subsequent hypertension, independent of HDP . [Conclusions] 20 wks BP may predict both HDP and subsequent hypertension 5 years post delivery.

Prenatal screening using the maternal serum markers alpha-fetoprotein, human chorionic gonadotropin, and unconjugated oestriol was investigated in a native Japanese population. Comparison with a Caucasian U.S. population revealed differences which led to modification of the generally used equations for risk calculations. Prenatal screening was shown to be clinically useful.

To study the diversity of the protocadherin family, the cDNA clones for a novel protocadherin were isolated by screening rat brain cDNA libraries with a cDNA fragment obtained by PCR, and some of the properties were then characterized. The overall structure of the protein defined by the clone is similar to that of previously identified protocadherins; however, the cytoplasmic domain is distinct from those of previously cloned protocadherins or any other protein sequences in the data bank. We named this protocad herin-3 (Pcdh3) since this is the third protocadherin of which the entire coding sequence has been determined. Most of the deduced amino acid sequences of other cDNA clones obtained by the screening show high homology with but are distinct from that of Pcdh3, indicating that most of these sequences correspond to homologous but different protocadherins. These results demonstrate that Pcdh3 and the protocadherins defined by these clones constitute a protocadherin subfamily. Chromosome mapping indicates that mouse Pcdh3 is located in a specific region of mouse chromosome 18, close to the location of previously cloned protocadherins, suggesting that various protocadherins form a cluster in this region. In situ hybridization results showed that Pcdh3 and its related proteins were expressed at various areas in brain. The expressed Pcdh3 protein from the cDNA in mouse L cells was about 100 kDa in molecular weight and was localized at cell-cell contact sites. In contrast to the classical cadherins, however, the expressed Pcdh3 was sensitive to trypsin even in the presence of Ca2+, and the transfectants did not show strong Ca(2+)-dependent cell aggregation activity. These results indicate the structural and possibly functional diversity of the protocadherin family and suggest a distinctive biological role for Pcdh3.

Cadherin-11 (cad-11) belongs to the cell adhesion type II cadherin family, which seems to have different functions from the classic cadherin family. This study shows the overall pattern of cad-11 gene expression during rat embryonic development, from the pregastrula to very late embryonic stage. Cad-11 is the first cadherin found to be highly expressed in the dispersed and migrating mesenchymal cells that originate from the neuroectodermal neural crest cells and from the pre-chordal and paraxial mesoderm. A burst of cad-11 expression appears during the epithelial to mesenchymal transition, as observed by sclerotome formation. Cad-11 mRNAs were present in all mesenchymal cells throughout the embryo, regardless of their embryonic origin. A proximo-distal and antero-posterior gradient of cad-11 expression is seen in the limb buds, genitalia, and tail. As development proceeds, while all epithelium are negative, cad-11 is present in all mesenchymal cells involved in various morphogenetic events, such as the mesenchyme condensations during chondrogenesis and in the formation of sclera, cornea, naris, palate and meninges. Cad-11 was strongly expressed in mesenchyme during lung or kidney branching morphogenesis or the many epithelium to mesenchyme inductions that operate in the nasal septum, skin, vibrissae, teeth and various glands. High levels of cad-11 transcripts were also found in the dispersed cells of the hyaloid plexus in the vitreous body and in the invading mesenchyme within the trabeculae of the outflow tract of the heart. Cad-11 is thus specific to the mesenchymal phenotype whatever the stage of embryonic development.

Elevated blood pressure (BP) at early or mid pregnancy is a known risk factor for pregnancy-induced hypertension (PIH). However, the association between BP changes during the first half of pregnancy and subsequent PIH development is unknown. We used changes in maternal BP between 16 and 20 weeks of gestation to evaluate the risk of PIH. A total of 976 pregnant women with BP estimations recorded before 16 weeks and at 20 weeks of gestation participated in this study. BPs were classified by the Japanese Society of Hypertension 2009 Hypertension Treatment Guidelines (JSH 2009). There was a significant trend for future PIH in women whose JSH 2009 BP class increased between 16 and 20 weeks of gestation, and the risk of PIH was highest among women whose BP was Class IV Hypertension (systolic BP≥140 mm Hg and/or diastolic BP≥90 mm Hg). The risk of PIH increased in women whose BPs shifted from Classes I Optimal (systolic BP120 mm Hg and diastolic BP80 mm Hg) and II Normal (systolic BP 120-129 mm Hg and/or diastolic BP 80-84 mm Hg) before 16 weeks to Class III High-Normal (systolic BP 130-139 mm Hg and/or diastolic BP 85-89 mm Hg) at 20 weeks of gestation. These shifts in BP class were significantly correlated with the risk of PIH after adjustments for variables (P-value for trend0.05). Within JSH 2009 Classes I, II and III, a shift in BP from a low to a high class between 16 and 20 weeks of gestation predicts the subsequent development of PIH.

In pregnant women with severe diabetes mellitus, intrauterine growth retardation (IUGR) may occur despite hyperglycemia. Uterine blood flow may be an important factor in this process. Using pregnant streptozotocin-induced diabetic rats (pregnant STZ-diabetic rats), we evaluated functional and morphological changes in the endothelium, as well as platelet thromboxane A2 (TXA) production and their influence on uterine blood flow. Decreased basal endothelial prostacyclin (PGI2) production as well as significant attenuation of the production response to stimulus was noted, suggesting an impairment of PGI2 production. In the platelet, TXA2 production was augmented, leading to a hypercoagulable state. Decreased myometrial blood flow was documented. Scanning electron microscopy (SEM) revealed definite morphological abnormality of the endothelial cells, probably affecting the permeability to molecules, a major endothelial function. Based on these data, the endothelial derangements suggested by SEM and arachidonic acid metabolism alterations may affect fetal growth in the severe pregnant diabetic through alterations in uterine blood flow.

For a parturient with breech presentation, an external cephalic version is sometimes done to enable vaginal delivery. Usually external cephalic version has been done without anesthesiologist's management. However there have been several reports indicating a benefit of anesthesia for external cephalic version. We report successful case of external cephalic version under combined spinal-epidural anesthesia followed by vaginal delivery.

Chronic granulomatous disease (CGD) is a primary immunodeficiency characterized by recurrent infections with certain types of bacteria and fungi. Presented herein is the case of a 29 year old woman with CGD who suffered from bacteria-associated haemophagocytic syndrome and a septic pulmonary embolism following a uterine infection and sepsis, caused by Burkholderia cepacia complex.

Background A considerable effort is being spent in developing noninvasive prenatal DNA diagnostic procedures. We recently reported that nucleated erythrocytes (NRBCs) can be enriched from maternal blood by a galactose-specific lectin method. In the present study, to prove that fetal NRBCs are definitely present in maternal blood and are a good source for fetal genetic diagnosis, we evaluated methods for lectin enrichment and subsequent fluorescence in situ hybridization (FISH) analysis through fetal gender determination. Methods Peripheral blood samples were collected from pregnant women (median 15, range: 10–18 weeks). From the blood samples, NRBCs were enriched based on galactose-specific lectin method. After detecting them by their morphology, NRBCs are separated and taken in a new glass slide by micromanipulator. We analyzed fetal gender using X and Y-chromosome-specific FISH probes. The results were compared with fetal gender analysis using Y-chromosomal sequences in maternal plasma. Results The fetal gender analyses by FISH in 20 pregnant women were all in accordance with the results from maternal plasma analyses. It is confirmed that fetal NRBCs were present in maternal blood and that 30.4% of NRBCs in maternal blood were fetal in origin. Conclusion We have successfully carried out a noninvasive prenatal DNA diagnosis of fetal gender by using galactose-specific lectin method and subsequent FISH analysis. Copyright  2007 John Wiley & Sons, Ltd.

Objectives Hypertensive disease in pregnancy (HDP) was revealed as a risk of subsequent hypertension five years after index delivery in our previous study. However the association between the postnatal blood pressure (BP) and subsequent hypertension is unclear. We investigate the association between the BP one month after index delivery and the risk of subsequent hypertension. Methods The study subjects were recruited from“Seiiku cohort study”at the National Center for Child Health and Development in Tokyo from 2003 to 2005. After that period, women suffering from HDP at high risk of suffering cardiovascular diseases were given follow-up examinations and are included in this study. They were invited to undergo a physical examination five years after delivery and 753 women completed the analysis. The influence of postpartum BP on subsequent hypertension was assessed by a multiple logistic regression analysis. The odds ratio for subsequent hypertension was adjusted by age, BMI, familial history of hypertension, kidney disease and salt intake. [Criteria] HDP was defined as sBP ⩾140 mmHg or dBP ⩾90 mmHg from 20 weeks’ gestation to 12 weeks postpartum w/o proteinuria. Hypertension at physical examination was defined as average sBP ⩾140 mmHg or average dBP ⩾90 mmHg or treatment with antihypertensive agents. [Exclusion criteria] The women with chronic hypertension before pregnancy and those pregnant and nursing when the physical examination was done. Results There were 37 HDP cases and 23 cases of subsequent hypertension (HDP:5, normotensive control:18). The odds ratio for subsequent hypertension is sBP:1.11 (95%CI: 1.04–1.17), dBP:1.13 (1.05–1.22) and mBP:1.15 (1.06–1.24). Despite adding HDP to confounding factors, the odds ratio is still sBP:1.10 (1.04–1.17), dBP:1.12 (1.04–1.22) and mBP:1.14 (1.05–1.23). Conclusions Postnatal BP may predict subsequent hypertension 5 years post-delivery, independent HDP. Disclosures A. Mito: None. N. Arata: None. D. Qiu: None. N. Sakamoto: None. Y. Ohya: None. A. Murashima: None. A. Ichihara: None. M. Kitagawa: None.

Based on 9350 pregnant Japanese women who were screened by serum triple-marker determination, accuracy of predicted risk for Down syndrome was examined using 24 Down syndrome cases detected either prenatally or postnatally. The correlation is statistically very high (r = 0.98) between the predicted risks and the prevalence of Down syndrome cases. Here we emphasize that this could be accomplished only by an extensive follow-up study, implemented in our prospective intervention programme.

We performed in vivo and in vitro adhesion experiments on the use of photocrosslinkable chitosan (PC) for healing of the myometrium and the amniotic membrane to test its usefulness in the perinatal field.To determine the adhesion conditions of PC in the healing of the myometrium and amniotic membrane, we applied two concentrations (30 and 50 mg/mL) of PC to the postpartum amniotic membrane, and radiated it with ultraviolet (UV) light for 10, 20, 30 or 60 s. Next, we created incisions of three sizes (pinhole, 5 mm and 10 mm), and performed PC adhesion followed by pressure application experiments. Subsequently, we laparotomized 12-day-pregnant rats under anesthesia, created a 5-mm incision, applied PC and closed the abdomen. We removed the uterus at 20 days of pregnancy, and histologically examined the site of PC application.The optimal adhesion conditions of PC were 50 mg/mL, and the optimal time of UV irradiation was 30 s or longer. In pressure application experiments, the bonding strength of PC was so high that even 10-mm incisions withstood a mean pressure of 92 mmH2O. The bonding strength of PC was significantly higher than that of fibrin glue that was tested in parallel. Histological examination of the rat myometrium showed a mild foreign body reaction with no inflammatory findings, confirming the usefulness of PC.PC is a wound-healing drug, an improvement on the naturally occurring polysaccharide chitosan, and is safer and has a higher bonding strength than biological adhesives including fibrin, making it promising for fetal surgical and perinatal applications.

We previously reported the separation and recovery of nucleated red blood cells (NRBCs) in maternal blood using the lectin method. In the present study, we verified the lectin method and investigated the appearance of NRBCs during pregnancy. For the concentration of lectin soy bean agglutinin, 7 mL of maternal peripheral blood was collected from 20 subjects, and the relative fluorescence intensity was measured using flowcytometry; 50 mg/mL, used in previous studies, was the optimal concentration. The number of cells recovered at each step of the lectin method was also investigated by FACS using fluorescence-labeled CD11a and CD33, and the results showed the usefulness of the method. Next, 7 mL of maternal peripheral blood was collected from 292 women with a normal single pregnancy (389 specimens), and NRBCs were separated and recovered using the lectin method. NRBCs slightly increased over the course of pregnancy (y = 4.29x + 5.03, r2 = 0.11). When blood was collected multiple times in the same subjects, NRBCs increased in 63 of 77 subjects (83.1%, percent change: 2.4 +/- 19.0). No NRBCs were recovered in 17 subjects (4.7%). Regarding the relationship between fetomaternal disorders and the frequency of NRBCs, 89.4 +/- 92.6 cells appeared per 10 mL of maternal blood in the normal group, but NRBCs increased in patients with 18 trisomy, placenta previa, pre-eclampsia, intrauterine fetal death, and 21 trisomy. NRBC examination may play an assisting role not only in fetal diagnosis but also in fetomaternal diagnosis.

Mirror syndrome is a preeclampsia-like disease first described in a case of severe hydrops fetalis caused by rhesus isoimmunization, later reported in some cases of nonimmunological fetal hydrops. Twin-twin transfusion syndrome (TTTS) is a severe complication associated with monochorionic pregnancies, in particular, severe TTTS with one hydropic fetus leading to a poor prognosis. We report here a case of mirror syndrome that occurred after selective fetoscopic laser photocoagulation in severe TTTS at 24 weeks’ gestation.

Fetal nucleated cells in maternal peripheral blood are a non-invasive source of fetal DNA for prenatal genetic diagnosis. However, the number of fetal cells present in maternal peripheral blood is very small. Therefore, fetal cell enrichment is generally considered necessary to allow detection and subsequent genetic analysis of the rare fetal cells. In the study presented here, we performed fetal cell separation from maternal blood using galactose-specific lectin to concentrate fetal nucleated red blood cells (FNRBCs), and attempted paternal diagnosis using polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP). Fetal cell separation was performed using galactose-specific lectin on a PV-MeA coated slide. Twenty cells consisting of an NRBC and its surrounding 19 maternal cells were collected using laser microdissection for stable DNA amplification. DNA analysis was performed using three sequence tagged site markers (D13S270, D17S5, and D18S474) by PCR-SSCP. All seven cases were informative because they showed heterozygosity at least one locus in D13S270, D17S5, or D18S474, and paternal-specific bands were detected in all cases. These results suggest that our proposed lectin-laser-micromanipulation-PCR-SSCP method may contribute to the development of prenatal diagnosis.

Protocadherin (Pcad) is a group of molecules obtained by polymerase chain reaction (PCR) utilizing the sequence that is well preserved in the extracellular domain of cadherin. Sano et al. analyzed Pcad (PC42,43) that had been cloned from rats, and found that it basically had homology to cadherin, but contained more than six cadherin repeats with a completely different intracellular domains (Sano et al. 1993). In the present study, of the Pcad (Pcad-1,2) cloned from a human cDNA library, as-yet-unspecified Pcad-2 was analyzed for expression in the human fetal central nervous system (CNS). Northern blot analysis of adult human tissue showed that Pcad-2 was expressed in the brain and the placenta, and that Pcad-2 mRNA was expressed in actively dividing neural tumor cell lines. Monoclonal antibodies against Pcad-2 were then made, and the CNS of fetuses were immunohistochemically stained. The expression was hardly visible at the 6th week of pregnancy, and began to become visible along the nerve fiber in the brain stem at the 8th week, and spread over the entire brain at the 11th week. At the 18th week, however, expression in the nerve fascicles, which had been visible by that time, was no longer visible or had decreased. These results suggest that Pcad-2 appears relatively early in the critical stage of development of the fetal CNS, and is involved in the induction, fasciculation, and extension of axons.

A fetal diagnostic method that is without risk to the embryo has been long awaited in the field of gene diagnosis. Establishment of non-invasive fetal diagnosis using maternal peripheral blood will greatly contribute to perinatal medical care. The lectin method that we have studied and developed selectively recovers nucleated red blood cells (NRBC) among fetal cells mixed in maternal peripheral blood. Maternal blood, 7 mL, was collected with ethylene diamine tetraacetic acid (EDTA) treatment in each week of gestation and subjected to preliminary concentration by density centrifugation (Histopaque [Histopaque-1077; Sigma Diagnostics, MO, USA], specific gravity: 1.095), and NRBC were separated and collected on slide glasses by the lectin method (soybean agglutinin [SBA]: 50 microg/mL). To investigate selective adhesion of the erythrocyte fraction, the SBA concentration was set to 50 microg/mL, and the cells were labeled with CD11a and CD33 (anti-white blood cell antibodies) and investigated by flowcytometry. Erythrocytes adhered at a high rate (87.0 +/- 9.7%) while the adhesion rates of granulocytes, monocytes, and lymphocytes were low, confirming the usefulness of the method for separation and recovery of the erythrocyte fraction. When recovery of NRBC was investigated using this method, a mean of 6.57 +/- 7.12 cells were recovered from 1 mL of maternal blood (May-Gluuwald-Giemsa stain). The number of recovered NRBC increased slowly with pregnancy, but differences were not significant. To confirm that recovered NRBC were derived from the fetus, NRBC were recovered by the lectin method in four patients suspected of 18 trisomy by echography and analyzed by fluorescence in situ hybridization. Three hybridization signals were detected in NRBC at a high frequency, showing that most cells were derived from the fetus and thus, fetal diagnosis may be possible. Since the procedure of the lectin method we have developed is simple, and high concentration efficiency can be obtained at a low cost, it may be clinically applicable.

Pregnancy-induced hypertension (PIH) is associated with increased risk for cardiovascular diseases later in life.To assess the prevalence of subsequent hypertension and other life-style diseases five years after delivery in women who experienced pregnancy- induced hypertension.A total of 1527 women who delivered singletons were registered at the National Center for Child Health and Development and Showa University Hospital Mother and child health center of integrated perinatal period between 2003 and 2005. After five years, these women were invited to participate in this study by mail, and 816 women completed the analysis. The women visited our hospital and underwent a medical examination. Women who were pregnant and nursing at the time when the physical examination was conducted were excluded from this survey. The outcomes assessed included the prevalences of hypertension, diabetes, and dyslipidemia.The number of PIH cases was 27 (3.3%: PIH group), whereas 787 women were used as controls subjects. The mean blood pressure five years after delivery was higher in the PIH group than in the control group (91.6±15.5mmHg vs 82.4±8.8mmHg, respectively; p0.001), and the prevalence of hypertension five years after labor was 18.5% in the PIH group and 2.9% in the controls (odds ratio UOR Y=6.2; 95% confidence interval (CI)=2.2-17.5; p=0.003). Moreover, regarding high-normal blood pressure (130/85mmHg), the prevalence was 33.4% in the PIH group and 6.1% in the control subjects (OR=7.2; 95% CI=3.1-16.3; p=0.003). No differences in the prevalences of subsequent diabetes or dyslipidemia were observed.Five years after the index pregnancy, women who experienced PIH exhibit an increased risk for subsequent hypertension. Therefore, the blood pressure of women with history of PIH should be regularly monitored after delivery.

Objectives: Studies indicate that Helicobacter pylori (HP) infection is closely related to gastric mucosa lesions and well-differentiated gastric cancer. In Japan, the HP-positive rate in childhood is 5–6%, which is similar to other developed countries, and in regard to the infection route, oral infection is considered important. To our knowledge there have been no reports on mother-to-child transmission and in this study we investigated maternal HP infection status to determine the potential of mother-to-child transmission in the perinatal period. Methods: After obtaining informed consent from 1,588 pregnant women, mother's blood and cord blood were collected at delivery to measure HP antibody (Helico-G). Gastric contents from the neonates were cultured to isolate H. pylori (Skirrow medium). Vaginal discharge (73 women) and dental plaque scraping swabs (48 women) were collected before delivery, and milk (66 women) was collected after delivery from 212 HP antibody-positive pregnant women to detect H. pylori by PCR. Results: The HP antibody-positive rate for the pregnant women was 29.2%. H. pylori was not detected in the vaginal discharge from HP antibody-positive pregnant women, but dental plaque scraping swabs from 4 women and milk from 4 women was positive. Conclusion: We considered that vertical infection during pregnancy or at delivery is unlikely as a route of mother-to-child HP antibody infection. However, horizontal infection through breast-feeding may occur.

Objective: The purpose of this study was to assess the accuracy, informative rate, detection rate, and clinical utility of prenatal interphase fluorescence in situ hybridization (FISH) analysis of amniotic fluid samples from Japanese women. Methods:Amniotic fluid specimens from 2,639 Japanese women were received for prenatal interphase FISH and chromosome analysis. A questionnaire was designed to evaluate FISH clinical utility by collaboration sites. Results:Based on 2,319 tested samples, the accuracy (100%), informative (94%), and detection (87.6%) rates were all high. The accuracy (100%), informative (90.2%), and detection (90.0%) rates were also remarkable in third-trimester pregnancies. We perceive significant advantages from this test regarding medical management and patient satisfaction. Conclusions:This novel report shows that in Japan prenatal interphase FISH testing is highly informative and accurate, not only in second-trimester pregnancies but also in third-trimester pregnancies. This test provides advantages to both physicians and patients, provided that its capabilities and limitations are understood.