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The objective of our study was to compare and consider reference values of fetal atrioventricular (AV) intervals as measured by four different pulsed Doppler wave techniques (left ventricular inflow/outflow [LV in/out], pulmonary vein/pulmonary artery [PV/PA], innominate vein/ascending aorta [InnV/AA], and supra vena cava/ascending aorta [SVC/AA]) in pregnant women with anti-SSA/Ro antibodies.Between March 2014 and September 2020, 52 pregnant women with anti-SSA antibodies were enrolled. No bradyarrhythmia was observed in the group. A pulsed Doppler examination of the fetal heart was performed to obtain measurements of the mechanical Doppler AV interval. Doppler measurements were performed using four methods: LV in/out, PV/PA, InnV/AA, and SVC/AA. A statistical analysis was performed to examine the mean, standard deviation, significant difference, and correlation of the four methods. The detection rate of each method was also calculated.There was no significant difference in the AV intervals between any of the four methods. There was also a positive correlation in the AV intervals of each of the four methods. The fetal heart rate and AV interval showed no correlation. The gestational age and AV interval also showed no correlation. The detection rate was highest for LV in/out (62.6%, 95% confidence interval: 56.5-68.4).All four pulsed Doppler methods are useful for measuring AV intervals. The most practical method is LV in/out.

AIM The association between methylenetetrahydrofolate reductase (MTHFR) C677T polymorphism and intervillous and decidual pathology in patients with pregnancy loss was investigated. METHODS We performed a cross-sectional study on 243 patients presenting with pregnancy loss for the degree of intervillous fibrin and thrombosis (IT), and decidual fibrin and thrombosis (DT) and determined their MTHFR C677T genotypes. Overall differences in age, body mass index (BMI), gravidity, parity, number of pregnancy losses and gestational period when the pathologic samples were obtained, also were determined. RESULTS There were no significant differences in age, BMI, gravidity, parity, number of pregnancy losses and gestational period, relative to MTHFR C677T genotype (TT vs CT vs CC). There were significantly more T allele carriers and TT genotype patients among patients with severe IT (odds ratio [OR] 1.653, P = 0.033 and OR 2.246, P = 0.032, respectively) and those with severe IT and decidual thrombosis (OR 2.602, P = 0.012 and OR 3.375, P = 0.035, respectively). The CC genotype was protective against the four studied pathologic grades. CONCLUSION To our knowledge, this is the first study showing that the MTHFR C677T TT genotype and T allele are associated with severe intervillous and decidual pathologies in patients with pregnancy loss. Differences in pathologic grades of MTHFR C677T TT genotype could support the hypothesis that further periconceptional treatment for pregnancy loss could be customized depending on single nucleotide polymorphisms.

Ureaplasma spp. cause several disorders, such as nongonococcal urethritis, miscarriage, and preterm delivery with lung infections in neonates, characterized by pathological chorioamnionitis in the placenta. Although reports on antibiotic resistance in Ureaplasma are on the rise, reports on quinolone-resistant Ureaplasma infections in Japan are limited. The purpose of this study was to determine susceptibilities to five quinolones of Ureaplasma urealyticum and Ureaplasma parvum isolated from perinatal samples in Japan and to characterize the quinolone resistance-determining regions in the gyrA , gyrB , parC , and parE genes. Out of 28 clinical Ureaplasma strains, we isolated 9 with high MICs of quinolones and found a single parC gene mutation, resulting in the change S83L. Among 158 samples, the ParC S83L mutation was found in 37 samples (23.4%), including 1 sample harboring a ParC S83L–GyrB P462S double mutant. Novel mutations of ureaplasmal ParC (S83W and S84P) were independently found in one of the samples. Homology modeling of the ParC S83W mutant suggested steric hindrance of the quinolone-binding pocket (QBP), and de novo prediction of peptide structures revealed that the ParC S84P may break/kink the formation of the α4 helix in the QBP. Further investigations are required to unravel the extent and mechanism of antibiotic resistance of Ureaplasma spp. in Japan.

Antiphospholipid syndrome (APS) is the most important treatable cause of recurrent pregnancy loss. The live birth rate is limited to only 70–80% in patients with APS undergoing established anticoagulant therapy. Lupus anticoagulant (LA), but not anticardiolipin antibody (aCL), was found to predict adverse pregnancy outcome. Recent genome-wide association studies (GWAS) of APS focusing on aCL have shown that several molecules may be involved. This is the first GWAS for obstetric APS focusing on LA. A GWAS was performed to compare 115 Japanese patients with obstetric APS, diagnosed according to criteria of the International Congress on APS, and 419 healthy individuals. Allele or genotype frequencies were compared in a total of 426 344 single-nucleotide polymorphisms (SNPs). Imputation analyses were also performed for the candidate regions detected by the GWAS. One SNP (rs2288493) located on the 3′-UTR of TSHR showed an experiment-wide significant APS association (P=7.85E-08, OR=6.18) under a recessive model after Bonferroni correction considering the number of analyzed SNPs. Another SNP (rs79154414) located around the C1D showed a genome-wide significant APS association (P=4.84E-08, OR=6.20) under an allelic model after applying the SNP imputation. Our findings demonstrate that a specific genotype of TSHR and C1D genes can be a risk factor for obstetric APS.

Deep vein thrombosis (DVT) is a serious pregnancy-related complication. Recent studies indicate that the genetic background for DVT differs with ethnicity. In our study, we enrolled 18 consecutive Japanese patients who had developed DVT during pregnancy and postpartum. We performed a genetic analysis of three candidate genes for DVT, protein S, protein C and antithrombin, in these patients. We found that four patients had missense mutations in the protein S gene, including the K196E mutation in two patients, the L446P mutation in one patient, and the D79Y and T630I mutations in one patient, as well as one patient with the C147Y mutation in the protein C gene. All five patients with genetic mutations had DVT in their first two trimesters. Nine of the patients without genetic mutations developed DVT in the first two trimesters, and four in the postpartum period. Thus, genetic mutations in the protein S gene were predominant in pregnant Japanese DVT women, and DVT in pregnant women with genetic mutations occurred more frequently at the early stage of pregnancy than postpartum. Considering the rapid decrease in protein S activity during pregnancy, we may need to assess thrombophilia in women before pregnancy.

Recent findings have raised the possibility that polymorphisms within the annexin A5 gene (ANXA5) promoter contribute to the etiology of recurrent pregnancy loss (RPL). In our present study, 243 Japanese women who had suffered more than three fetal losses and a group of 119 fertile controls were genotyped for four ANXA5 gene promoter single-nucleotide polymorphisms (SNPs; SNP1-4: g.-467G .A, g.-448A.C, g.-422T.C, g.-373G.A) previously reported to be associated with this disorder. An additional two SNPs located within the 5 ' -untranslated region of the ANXA5 (SNP5 and 6: g.-302T.G, g.-1C.T) were also evaluated. Our case -control study revealed that the minor allele was significantly more frequent in the RPL group than controls for all six of these SNPs, among which SNP5 showed the highest significance (P ¼ 0.002). As with the M2 haplotype for SNP1-4 (A-C-C-A) for a western population in previous reports, a haplotype comprising all of the minor alleles for SNP1-6 (A-C-C-A-G-T), the third major haplotype in the Japanese population, showed a significantly higher frequency in our current RPL subjects than in controls (P ¼ 0.025). In addition, the second major haplotype (G-A-T-G-G-C) was found to confer a significant risk of RPL (P ¼ 0.036), implicating SNP5 as a major risk determinant for this disease. Our present findings support the hypothesis that genomic variations within the ANXA5 gene upstream region impact upon the disease susceptibility to RPL. Our data indicate that SNP5 is a novel risk factor for this disease in the Japanese population.

Ureaplasma parvum serovar 3 strain, OMC-P162, was isolated from the human placenta of a preterm delivery at 26 weeks’ gestation. In this study, we sequenced the complete genome of OMC-P162 and compared it with other serovar 3 strains isolated from patients with different clinical conditions. Ten unique genes in OMC-P162, five of which encoded for hypothetical proteins, were identified. Of these, genes UPV_229 and UPV_230 formed an operon whose open reading frames were predicted to code for a DNA methyltransferase and a hypothetical protein, respectively. DNA modification analysis of the OMC-P162 genome identified N4-methylcytosine (m4C) and N6-methyladenine (m6A), but not 5-methylocytosine (m5C). UPV230 recombinant protein displayed endonuclease activity and recognized the CATG sequence, resulting in a blunt cut between A and T. This restriction enzyme activity was identical to that of the cultivated OMC-P162 strain, suggesting that this restriction enzyme was naturally expressed in OMC-P162. We designated this enzyme as UpaP162. Treatment of pT7Blue plasmid with recombinant protein UPV229 completely blocked UpaP162 restriction enzyme activity. These results suggest that the UPV_229 and UPV_230 genes act as a type II restriction-modification system in Ureaplasma OMC-P162.

Information concerning the prognosis of subsequent pregnancies in patients with reciprocal translocations is limited. This study was performed to determine the percentage success rate with first pregnancies after ascertainment of a carrier status. A total of 2,382 couples with a history of two or more consecutive miscarriages were studied in multicenters. The prevalence of an abnormal chromosome in either partner was examined, and subsequent success rates were compared between cases with and without an abnormal karyotype in either partner. A total of 129 couples (5.4%) had an abnormal karyotype in one partner excluding inversion 9 in 44 men and in 85 women. Thus, 2,253 couples had a normal karyotype in both partner. Eighty-five (3.6%) had translocations, 13 being Robertsonian translocations. Twenty-nine of the 46 cases (63.0%) who became pregnant with reciprocal translocations in either partner experienced a live birth with natural conception. In contrast, 950 of 1,207 cases (78.7%) with normal chromosomes had successful live births, the difference being significant (P = 0.019). No infant with an unbalanced translocation was found in 29 cases of successful pregnancy following recurrent miscarriage. Pregnancy prognosis was worsened with either maternal or paternal reciprocal translocations. Explanation of the success rate with natural conception should be provided before the subsequent pregnancy after ascertainment of carrier status.

Context: Adiponectin (APN) concentration in umbilical cord serum is higher than that in adult serum. Except for the positive association between birth weight and cord APN concentration, little is known about the pathophysiological function of APN in fetal development. Objective: The objective of this study was to evaluate the relationship of cord serum APN and IGF-I concentrations with the development of the fetoplacental unit. Design and Methods: Umbilical cord serum APN and IGF-I concentrations were measured in term singleton deliveries (n = 94). The association of cord APN and IGF-I concentrations was evaluated in relation to fetal weight, placental weight, and fetoplacental (F/P) weight ratio. Results: Mean concentrations and sd of APN and IGF-I were 36.1 ± 14.0 μg/ml and 58.6 ± 27.0 ng/ml, respectively. Cord APN concentration was positively associated with F/P weight ratio (r = 0.375, P < 0.001) as well as fetal weight (r = 0.389, P < 0.001) but not placental weight. Cord IGF-I concentration was positively associated with fetal weight (r = 0.405, P < 0.001) and placental weight (r = 0.400, P < 0.001) but not F/P weight ratio. In multiregression analysis, only APN concentration resulted in a significant determinant of F/P weight ratio among variables (β = 0.376, P < 0.001). Conclusions: In cord hyperadiponectinemia, fetuses tend to be disproportionately larger for their placental weight and vice versa in cord hypoadiponectinemia. APN is shown to be the first biomarker positively associated with F/P weight ratio.

Here, we present the complete genome sequence of Ureaplasma parvum serovar 3, clinical strain SV3F4, isolated from a Japanese patient with a history of an infectious abortion.

When the concept of brain death is not widely accepted, the viability of renal allografts of non-heart-beating cadaveric donors is one of the serious concerns for transplant surgeons. At our center, 200 kidneys were harvested from cardiac arrest donors using the regional in situ cooling technique, and they were transplanted into recipients treated with cyclosporine (CS). The ages of donors ranged from 7 months to 67 years. At the bedside a specially designed double balloon catheter, 14F, was inserted into the aorta through the femoral artery just before or immediately after the cardiac arrest. A venous drainage tube was also placed in the vena cava. Following the cardiac arrest, both balloons of the aortic catheter were inflated, and regional in situ cooling with cold Lactate Ringer's solution started using the infusion pump at 20 mL/kg/min. In the OR, both kidneys were removed en bloc and preserved in Collins'type solution. They were then transplanted into 200 patients treated with CS and steroid. After the transplant operations, 33 patients (16.5%) had immediate renal function, but 14 grafts (7.0%) were not successful and the patients have never had renal function. When several factors such as donor age, warm ischemic time (WIT; 12.3 ± 14.1 minutes), in situ cooling time (IST; 78.1 ± 18.0 minutes) and total ischemic time (TIT; 619 ± 340 minutes) were associated with the post-transplant renal function, only the donor age had significant correlation both with the posttransplant dialysis period and lowest serum creatine level, as follows: 10.5 days ( ≤ 40 years) vs. 14.6 days (40 years); P0.05, and 1.16 mg/dL ( ≤ 30 years) vs. 1.83 mg/dL (50 years) P0.001, respectively. Our findings indicate; 1) Due to the in situ cooling technique, the renal grafts of non-heart-beating cadavers can be expected to have relatively good function in the CS-treated recipients; 2) donor age is instrumental in predicting post-transplant renal function as well as the duration of ATN; 3) WIT, 1ST and TIT have no association with the post-transplant renal function if the duration of renal ischemia is within the acceptable range.

OBJECTIVE We have already reported that A1C is elevated because of iron deficiency in late pregnancy among nondiabetic pregnant women. This report examined whether the same phenomenon is observed in pregnant women with diabetes. RESEARCH DESIGN AND METHODS This longitudinal study was conducted in 17 pregnant women with diabetes (20–35 weeks of pregnancy). A1C, serum glycated albumin, erythrocyte indexes, and iron metabolism indexes were measured. RESULTS A1C levels were significantly increased in late pregnancy, whereas serum glycated albumin showed no significant changes. Glycated albumin/A1C ratio, mean corpuscular hemoglobin, serum transferrin saturation, and serum ferritin were significantly decreased in late pregnancy. Serum transferrin saturation showed a significant positive correlation with glycated albumin/A1C ratio. CONCLUSIONS A1C levels, but not serum glycated albumin levels, are elevated in late pregnancy because of iron deficiency in diabetic women. Serum glycated albumin may offer an adequate marker for glycemic control during pregnancy.

Objective: To assess the possibility of preventing cardiac or cutaneous manifestations of neonatal lupus erythematosus or treating the fetus with congenital heart block by administering corticosteroid therapy to the mother. Methods: Eighty-seven offspring of 40 anti-Ro/SSA-positive mothers, followed up from 1979 to 1996, were evaluated. Autoantibodies against Ro/SSA and La/SSB antigens were detected by immunodiffusion and enzyme-linked immunosorbent assay. Results: None of 26 neonates whose mothers received corticosteroid maintenance therapy initiated before 16 weeks’ gestation demonstrated congenital heart block, whereas 15 of 61 neonates whose mothers received no corticosteroids during pregnancy or began receiving steroid therapy after 16 weeks’ gestation had congenital heart block. Complete congenital heart block, once developed, did not respond to corticosteroid treatment in utero. Four infants whose mothers received steroid treatment before 16 weeks’ gestation had skin lesions of neonatal lupus erythematosus. Conclusion: Once established, complete congenital heart block was irreversible and maternal corticosteroid therapy did not effectively prevent cutaneous lupus erythematosus. However, prenatal maintenance therapy with prednisolone or betamethasone given to the mother starting early in pregnancy (before 16 weeks’ gestation) might reduce the risk of developing antibody-mediated congenital heart block in the offspring.

An automated continuous measurement system for the monitoring of formaldehyde (HCHO) and acetaldehyde (CH3CHO) in the urban atmosphere was developed by using an annular diffusion scrubber in conjunction with a high-performance liquid chromatograph (HPLC). With this technique, atmospheric HCHO and CH3CHO were effectively collected by the annular diffusion scrubber which consists of a porous polytetrafluoroethylene (PTFE) tube disposed concentrically within a Pyrex-glass tube and a scrubbing solution. 2,4-Dinitrophenylhydrazine (DNPH) was selected as the scrubbing solution for collecting HCHO and CH3CHO, which are derivatized to 2,4-dinitrophenylhydrazone-formaldehyde (DNPH-HCHO) and 2,4-dinitrophenylhydrazone-acetaldehyde (DNPH-CH3CHO), respectively. An aliquot of the sample solution was automatically injected into an HPLC equipped with a semi-micro ODS column and a UV-VIS detector for separating and determining DNPH-HCHO and DNPH-CH3CHO. All the operations are sequenced by a programmable controller, and automated continuous measurements are performed with a typical temporal resolution of 1 h. The collection efficiencies of HCHO and CH3CHO were about 97% and 93%, respectively, at an air flow rate of 0.2 L/min. The lower detection limits (3σ of the blank hydrazones) of HCHO and CH3CHO were 0.05 ppbv and 0.10 ppbv, respectively, in the case of 12-L air sample volume. Analytical response of a standard solution of DNPH-HCHO and DNPH-CH3CHO by the HPLC during a 10-day continuous measurement was unchanged and the relative standard deviation (RSD) was < 1.0%. Interferences from O3 and NO2 were insignificant in this annular diffusion scrubber method. Both for HCHO and CH3CHO measurements, concentrations from this developed system well agreed with those measured by a DNPH Silica cartridge method.

環境水中の微量ほう素を分析するためにクロモトロープ酸の反応試薬を用いたポストカラム誘導体化法について検討し, フッ素, 塩素, 硝酸, 亜硝酸, 硫酸及びほう素などの陰イオン成分と同時に分析する方法を開発した。この方法はHPLCのカラムで陰イオン成分とほう素を分離し, フッ素, 塩素, 硝酸, 亜硝酸及び硫酸などの陰イオン成分を電気伝導度検出器で分析した後に, ほう素はクロモトロープ酸ポストカラム誘導体化法により励起波長330nm, 蛍光波長355nmの蛍光検出器で分析した。環境試料で本法, ICP発光分析法及び他法などとほう素の測定値について比較した。その結果, 本法とICPの測定値は一致した。水道水や河川水中などのフッ素, 塩素, 硝酸亜硝酸及び硫酸などの陰イオンとほう素の一斉分析法として有効と思われる。

Neonatal lupus erythematosus (NLE) is an antibody-mediated disorder of infants characterized by two major clinical manifestations; cutaneous lupus lesions and congenital heart block (CHB). The disease is associated with placentally transferred maternal anti-Ro/SSA and/or La/SSB antibodies. There is a tendency for the same disease expression to occur within a sibship. To reveal a possible association of class II MHC genes with maternal anti-Ro/SSA autoimmune responses and neonatal outcomes in NLE with a relatively homogeneous ethnic background, haplotype, and allele distributions were analyzed based on the PCR-RFLP results in 26 Japanese anti-Ro/SSA-positive mothers from three groups defined by neonatal outcomes. The results were as follows: (i) maternal HLA-DR5 haplotype DRB1*1101-DQA1*0501-DQB1*0301 and individual class II alIeles making up this haplotype were significantly associated with neonatal cutaneous lupus but not CHB. Conversely, maternal HLA-DQB1*0602 carried on HLA-DR2 haplotypes was associated with CHB but not cutaneous NLE; (ii) HLA-DQA1 alleles with glutamine at position 34 of the first domain, which have reportedly been associated with the autointmune responses to Ro/SSA antigens in other ethnic groups, were increased in the mothers of infants with cutaneous involvement; and (iii) there was no particular class II HLA profile that distinguished the disease manifestations in infants. These findings suggest that specific maternal MHC class II genes might correlate with specific neonatal outcomes in NLE.

Background: Neonatal lupus erythematosus (NLE) is a syndrome characterized by dermatitis and congenital heart block. The disease is mostly associated with transplacental passage of maternal anti-Ro(SS-A) or anti-La(SS-B) antibodies. Maternal HLA-DR3 and DQ2 alleles are associated with NLE in white and North American black populations. Objective: We sought evidence of a potential genetic disposition to NLE in mothers with a relatively homogeneous ethnic background. Methods: Class II human major histocompatibility complex HLA-DRB1, DQA1, DQB1, and DPB1 alleles were determined by polymerase chain reaction–restriction fragment length polymorphism in anti-Ro(SS-A)–positive mothers as well as in infants from seven Japanese families with siblings concordant or discordant for disease expression of NLE. Results: All seven mothers had two or three DQ alleles of DQA1 and DQB1 possessing specific amino acid residues, which are reportedly associated with anti-Ro(SS-A) autoantibody response in white and black populations. There was no class II HLA profile that distinguished disease manifestations of NLE in infants. Conclusion: The HLA class II allele associations with anti-Ro(SS-A) autoantibodies that have been noted in other ethnic groups were also found in Japanese anti-Ro(SS-A)–positive mothers whose infants had NLE, suggesting shared susceptibility factors across racial barriers in maternal predisposition to Ro(SS-A) autoimmune response. (J Am Acad Dermatol 1997;36:186-90.)

Nitric oxide (NO) production may be an important causal factor in hypertensive disorders during pregnancy. The plasma concentrations of NO2-(+) NO3-, stable metabolites of NO, were measured in 70 nonpregnant women, 323 normotensive pregnant women, 23 pregnant patients with preeclampsia, and 7 pregnant patients with essential hypertension. The normotensive women had higher plasma concentrations (30.0 +/- 0.6 mumol/l) than nonpregnant women (18.3 +/- 1.0 mumol/l; p0.0001). The plasma concentrations in the patients with preeclampsia (45.6 +/- 2.3 mumol/l) were higher than in the normotensive women (30.3 +/- 1.0 mumol/l; p0.0001) and were correlated with the systolic blood pressure (r = 0.442; p0.05). However, pregnant patients with underlying essential hypertension had significantly lower plasma concentrations (19.1 +/- 3.0 mumol/l; p0.005). These findings suggest that NO contributes to maternal vasodilation, the maintenance of uterine quiescence, and the pathogenesis and clinical features of hypertensive disorders during pregnancy.

Objective To examine endocrine states of mothers with normal 75-g oral glucose tolerance tests (GTTs) who gave birth to large for gestational age (LGA) neonates (group I) and to examine those neonates. Methods We examined plasma glucose levels and serum immunoreactive insulin responses after the 75-g oral GTT was given to group I mothers ( N = 34), mothers with an abnormal oral GTT who gave birth to LGA neonates (group II, N = 21), and those with normal oral GTTs having appropriate for gestational age neonates (group III, N = 173). We also examined the infants, checking neonatal birth weight, levels of immunoreactive insulin and C-peptide immunoreactivity in cord sera at birth and the lowest blood sugar level after birth to see if a correlation existed between them. Results Group I and II mothers showed higher titers in plasma glucose levels and remarkably enhanced ratios of 60 to 30-minute immunoreactive insulin values (immunoreactive insulin up-ratio) after load compared with those of group III mothers. Cord serum immunoreactive insulin and C-peptide immunoreactivity were significantly higher and the lowest blood sugar level was significantly reduced in group I and II neonates compared with those in group III. We observed a positive correlation between cord serum immunoreactive insulin, C-peptide immunoreactivity, and birth weight, but a negative correlation between cord serum immunoreactive insulin, birth weight, and the lowest blood sugar level in group I and II neonates (strongest tendency in group II), but not in group III neonates. Conculsion All of the abnormal carbohydrate metabolic responses in group I mothers and neonates may result in the promotion of growth in LGA fetuses similar to group II, but to a lesser extent. Identification of group I mothers by the immunoreactive insulin up-ratio after oral GTT will help predict the occurrence of LGA neonates and their possible hypoglycemia. (Obstet Gynecol 1995;85:242–9)

We report here two cases of recurrent miscarriages that were successfully treated with continuous intravenous administration of low molecular weight heparin (LMWH). One patient experienced 11 spontaneous abortions, and the other eight abortions. Previous treatments including prednisone, aspirin and mononuclear-cell immunization were all unsuccessful. They were negative for anticardiolipin antibodies and lupus anticoagulant, and had no inherited thrombophilic disorder. Intravenous administration of LMWH, 4800 units of dalteparin, was started as soon as the conception was confirmed, and was continued until 34 weeks of gestation. They were delivered of live born infants.

Retrospective examinations of 8,139 placentae were performed to clarify the relationship between placental disorders with massive intervillous fibrin deposition (MIFD) and intrauterine growth retardation (IUGR). Although the incidence of MIFD was low (0.4%), the small-for-date (SFD) birth rate in the MIFD group was significantly higher than that in the control group (62.9 vs. 8.3%; p0.001). Seventeen of 35 patients in the MIFD group had no clinical complications. MIFD itself was thought to be the main cause of IUGR in these patients. 78.4% of multiparae in the MIFD group have unsuccessful obstetrical histories such as intrauterine fetal death and fetal growth retardation. Four of 6 patients with a history of MIFD and SFD delivery in a previous pregnancy repeated the same episode. These data indicate that the MIFD recurrence rate in subsequent pregnancies must be high. Patients with a history of both SFD delivery and MIFD in previous pregnancies were defined as high-risk patients and they were given orally 30 mg of aspirin and 150 mg of dipyridamole daily and/or daily intravenous injection of 10,000 IU heparin during pregnancy. As a result, MIFD did not recur in all cases of the treated group and 87.5% (7/8) of the treated group could deliver approximate-for-date infants compared with 33.3% (2/6) of the untreated group (p0.05). These results indicate that anticoagulant and antiplatelet therapies are extremely effective for prevention of MIFD and IUGR due to MIFD.

Neonatal lupus erythematosus (NLE) is a rare disorder caused by the transplacental passage of maternal autoantibodies and manifests with characteristic skin eruption and/or congenital heart block. Anti-SS-A 52-kDa and 60-kDa antibodies are important serology markers for the diagnosis of NLE. However, women who have these antibodies do not always give birth to children with NLE. Therefore, we investigated the avidity (quality of the antibody) using urea derivative.The sera of 54 women and 19 umbilical cord blood specimens were measured using a commercial ELISA kit (MESACUP 52K SS-A/Ro and 60K SS-A/Ro, MBL). Avidity index (AI) was obtained using a reagent prepared according to the method described in the European patent EP 0875761 as well as in the Japanese patent application No. 10-121896.Mothers with infants demonstrating atrioventricular block (AB) showed higher antibody indices for 52-kDa protein than mothers of infants without AB (p=0.0145). Mothers with cutaneous NLE had higher antibody indices for 60-kDa protein than mothers without cutaneous NLE (p=0.0058). Mothers had higher antibody indices and AIs than fetuses. There was a positive correlation between the antibody index and AI. The level of AI was increased in patients demonstrating low platelet counts in umbilical cord blood, but high antibody indices did not always increase in relation to the low platelet counts in umbilical cord blood.Mothers with high antibody indices do not always give birth to children with NLE, even if the AIs of the mother are high. The fetus side, that is, the factor on the antigen side would be more important. However, we conjecture that anti-60-kDa antibody influences the development of cutaneous NLE, and that anti-52-kDa antibody influences the development of NLE with AB. The high AI may be related to more severe symptoms of NLE.

Surface modified molecularly imprinted polymers (SM-MIPs) for 17beta-estradiol (E2), utilizing 6-ketoecradiol as a pseudo template were prepared. MIPs for E2 were synthesized using 4-vinyl pyridine and ethylene dimethacrylate as a functional monomer and cross-linking agent, respectively. MIPs selectively retained E2 and provided excellent chromatographic resolution from interfering compounds inherent in river water sample matrices. Therefore, freshly prepared MIPs were applied to quantitative mass spectrometric (negative electrospray ionization mode) detection of low levels of E2 in river water sample. In order to pre-concentrate the target compound for HPLC analysis, column switching was coupled with a pretreatment column packed with the MIPs. The repeatability of actual determinations of river water sample, in which background E2 was not detected, spiked with 50 ng/L of E2 was 2.2% RSD with a detection limit and qualification limit of 1.8 and 5.4 ng/L, respectively. Surface modification of MIP particlefs packed in the pretreatment column provided selective affinity and on-line concentration of low levels of E2 while simultaneously eliminating sample matrix interference, resulting in a significant increase in sensitivity and reproducibility for liquid chromatography-mass spectrometry analysis of E2 in river water sample.

This study was undertaken to evaluate the hemostasis and coagulation profile of pregnant women receiving antiplatelet therapy with low-dose aspirin and dipyridamole for prevention of preeclampsia, intrauterine growth retardation, or pregnancy losses.Twenty-three pregnant women who received antiplatelet therapy with combined aspirin (40 mg.day(-1)) and dipyridamole (150 mg.day(-1)) were enrolled in the study. Platelet aggregation and coagulation tests were performed before the start of aspirin and dipyridamole, during medication, and at 3 days and 6 days after cessation of medication.Collagen-induced platelet aggregation was decreased during medication (25 +/- 26%, P0.001) and at 3 days after cessation of medication (46 +/- 35%, P0.001) compared with that before the start of medication (89 +/- 7%). ADP-induced platelet aggregation was decreased during medication compared with that before medication (66 +/- 18% vs 92 +/- 7%, P0.001). The platelet count, prothrombin time, activated partial thromboplastin time, bleeding time, and levels of fibrinogen and antithrombin III did not change over time. The blood loss of these patients during vaginal delivery and cesarean section did not differ from that of normal women during vaginal delivery and repeat cesarean section, respectively.At the doses used in this study, aspirin and dipyridamole inhibited platelet aggregation for up to 3 days after cessation of medication. This abnormality of aggregation was not detected by the bleeding time and was not associated with clinically abnormal bleeding.

We report here two cases of recurrent miscarriages that were successfully treated with continuous intravenous administration of low molecular weight heparin (LMWH). One patient experienced 11 spontaneous abortions, and the other eight abortions. Previous treatments including prednisone, aspirin and mononuclear-cell immunization were all unsuccessful. They were negative for anticardiolipin antibodies and lupus anticoagulant, and had no inherited thrombophilic disorder. Intravenous administration of LMWH, 4800 units of dalteparin, was started as soon as the conception was confirmed, and was continued until 34 weeks of gestation. They were delivered of live born infants.

Objective To investigate HLA class II allele associations with autoantibody responses to Ro/SS-A and La/SS-B among Japanese subjects. Methods Haplotype and allele distributions, along with molecular polymorphisms, of HLA class II genes were analyzed by polymerase chain reaction-restriction fragment length polymorphism in 41 Japanese women with precipitating autoantibodies to Ro/SS-A and/or La/SS-B. Results Among women with both Ro/SS-A and La/SS-B antibodies, the HLA class II haplotype DRB1*08032/DQA1*0103/DQB1*0601 and DRB1*08032 allele showed significantly increased frequencies compared with patients with anti-Ro/SS-A alone or with normal controls. All women with both anti-Ro/SS-A and anti-La/SS-B, but not those with anti-Ro/SS-A alone, carried DRB1 alleles that shared the same amino acid residues at positions 14-31 and 71 of the hypervariable regions of the DRB1 chain. All anti-Ro/SS-A positive women carried 1 or 2 alleles of DQB1*06 and DQB1*03 subtypes that shared the same amino acid residues at positions 71-77 of the DQB1 chain. HLA class II allele distributions did not differ among 3 anti-Ro/SS-A positive groups with different disease expressions, i.e., patients with systemic lupus erythematosus, patients with primary Sjogren's syndrome, and women with no apparent symptoms of rheumatic disease. Conclusion HLA class II allele distribution differ among anti-Ro/SS-A positive subjects according to the presence or absence of coexisting anti-La/SS-B antibodies, but not according to disease expression. Our findings suggest that different HLA class II molecules might control the development of anti-Ro/SS-A and/or anti-La/SS-B antibodies in the autoimmune response to the Ro/SS-A-La/SS-B complex.

Neonatal lupus erythematosus (NLE) is a rare disorder of neonates characterized by two major clinical manifestations: congenital heart block and cutaneous lupus lesions. The disease is associated with placentally transferred maternal anti-Ro/SSA and/or La/SSB antibodies. To clarify possible class II HLA associations with maternal autoantibody responses, haplotypic and allelic distributions, along with the polymorphism of the MHC class II HLA alleles, were analyzed based on PCR-RFLP results in 25 Japanese mothers of two groups defined by precipitating autoantibody profiles. Among mothers with both anti-Ro/SSA and anti-La/SSB antibodies, but not those with anti-Ro/SSA alone, the class II haplotypes DRB1*1101-DQA1*0501-DQB1*0301 and DRB1*08032-DQA1*0103-DQB1*0601 as well as individual class II alleles DRB1*1101, DRB1*08032 and DQB1*0301 showed significantly increased frequencies compared to those in normal controls. All anti-Ro/SSA and anti-La/SSB positive mothers carried DRB1 alleles that shared the same amino acid residues at positions 14-31 and 71 of the DRB1 chain. These mothers also carried homozygous or heterozygous DQ6 and DQ3 alleles that shared the same amino acid residues at positions 27-36 and 71-77 of hypervariable regions of the DQB1 chain. Furthermore, all mothers with both anti-Ro/SSA and anti-La/SSB were homozygous for DPB1*0501. Nine of 10 anti-Ro/SSA and anti-La/SSB-positive mothers, but only 6 of 15 mothers with anti-Ro/SSA alone, had affected infants. Thus, our findings suggest that there may be immunogenetic differences among mothers according to their autoantibody profiles, and that mothers with both anti-Ro/SSA and anti-La/SSB are more likely to have infants with NLE than mothers with anti-Ro/SSA alone.

Recently, an unstable DNA fragment specific to myotonic dystrophy (MyD) was discovered. In affected individuals, a DNA fragment is found that is larger than in normal siblings. Our objectives were to show whether the results of DNA analysis agree with the disease severity and prognosis in congenital myotonic dystrophy (CMyD) by DNA analysis. We investigated three pregnancies (two studied retrospectively) in three families. We genotyped the family members with the Southern blots and the polymerase chain reaction (PCR) analysis. In one case a prenatal diagnosis was carried out using chorionic villus sampling. This report also presents the three cases of affected mothers and CMyD babies with their growth courses. We clarify four main problems in CMyD, namely, respiratory distress, delayed motor development, feeding difficulty, and delayed mental development. The allele size in the range of 10 to 13 kb tended to be present as the adult form of MyD, and 14 to 15 kb as the CMyD. The three CMyD cases whose alleles size in the range of 14 to 15 kb showed various forms of disease and prognosis. We reached the following conclusions: the disease severity and prognosis in babies with CMyD did not correlate with the result of DNA analysis. The DNA analysis is a useful test for prenatal diagnosis. However, it is impossible to predict the disease severity and prognosis in babies with CMyD.

The levels of CH50, the complement components, C1q, C4, and C3, C3 degradation fragments, and factor B were determined in the cord blood of 128 newborn infants. The levels of C3, C4, and C3d3 (an index of chronic in vivo complement activation) were clearly lower in the 28 infants with respiratory distress syndrome (RDS) than in the infants with other lung diseases or with normal lungs. CH50 and factor B levels were also low in RDS. Levels of C1q and other serum components in RDS infants were similar to the average levels in other infants without RDS at a corresponding gestational age. Lung surfactant is synthesized in alveolar type 2 cells, in which the complement components C4, C3, and factor B, but not C1q, have been reported to be synthesized. It seems possible that common factors regulate the synthesis of some complement components and surfactant.

The development of the complement system was studied by quantitation of total hemolytic complement activity (CH50), C1q, C3, C4, and C3 split product (C3d) in cord plasma of nine human fetuses (17-22 weeks of gestation), 110 preterm (24-36 weeks of gestation) and 30 term neonates. The complement levels were analyzed in relation to various illnesses of preterm infants. Histological examination of the placenta revealed a higher incidence of amnionitis in the placenta of less than 34 gestational weeks. In cases without amnionitis, there were significant correlations between complement levels and gestational age. In cases with amnionitis, the complement system was activated even in infants of less than 28 weeks gestation. The complement levels correlated with the extent of the inflammation in the placentas and umbilical cords except for C1q. In infants with Wilson-Mikity syndrome, complement levels other than C1q were significantly elevated in comparison with those of infants with respiratory distress syndrome. In the group of preterm infants without amnionitis, no differences were found between infants with intrauterine growth retardation and those with growth appropriate for gestational age.

コンドロイチン硫酸の高速液体クロマトグラフィーによる分子量分布の測定と定量につき検討を加えた.ジオール型シリカゲルカラム(ボアサイズ300Å,粒子径5μm,500×7.9mm i.d.)と移動相としてメタノール-0.1M塩化カリウム水溶液(1:1)を用いた場合,良好な結果を得た.なお,分子量校正用の標準品としてボリエチレングリコールを用いた。本分析法を注射剤及び点眼剤のコンドロイチン硫酸に応用したところ,定量精度1%以内,分子量分布の測定精度2%以内で測定ができた.測定時間が30分以内で比較的短いことから,医薬品中のコソドロイチン硫酸の品質管理に本法が適用できるものと考える.

Tryptic hydrolyzates of normal and abnormal human globin chains were analyzed with mass spectrometry, using field desorption ionization technique. All the peptides, including cote peptides, were detected as protonated molecular ions in field desorption mass spectra. Since the technique makes it possible to determine the mass number of each peptide, it is very useful for structural analysis of human hemoglobin variants, even those with electrophoretically and chromatographically silent mutations.

A simple system for simultaneous fluorescence detection of primary and secondary amines has been developed. The optimum conditions for fluorescing secondary amines in the two-stage reaction technique using sodium hypochlorite and o-phthalaldehyde have been studied, and then the switching flow method (sodium hypochlorite is added only to secondary amines) and the non-switching flow method (sodium hypochlorite is added to all the amino acids) have been compared. It was concluded that the non-switching flow method ensures high efficiency and reliability. This newly-developed system was applied to the detection of amino acids and gave satisfactory results.

A grand-scale mass spectrometer with high mass resolution and high transmission was employed for the analysis of haemoglobin variant. Two variants were isolated from a haemolysate by chromatography. Secondary ion mass spectrometry of complex peptide mixtures derived from these variants precisely determined the molecular weight of abnormal peptides. The molecular weight, 2857.4 and 2858.4, indicated the amino acid substitutions of asparagine and aspartic acid, respectively, for lysine at position 82 of beta globin chain. The mutations had been reported in haemoglobin Providence.