Naohiro Tezuka

Incidence of apoptotic bodies in membrana granulosa of the patients participating in an in vitro fertilization program.

OBJECTIVE To investigate the incidence of apoptotic bodies in mural granulosa cell masses and cumulus cell masses. DESIGN Nonrandomized, prospective study. SETTING Department of Obstetrics and Gynecology, Yamagata University School of Medicine, Yamagata, Japan. PATIENT(S) One hundred twenty-nine normally ovulating women underwent ovulation induction for IVF-ET with GnRH analogue (GnRH-a) and gonadotropins. INTERVENTION(S) Patients underwent follicle aspiration after the administration of hCG. MAIN OUTCOME MEASURE(S) The nuclei of recovered granulosa cells were examined by fluorescence microscopy and the incidence of apoptotic bodies was tabulated. RESULT(S) The incidence of apoptotic bodies was significantly higher in mural granulosa cell masses than in cumulus cell masses in the entire group of 129 patients. Both incidence of apoptotic bodies of mural granulosa cell masses and cumulus cell masses were significantly higher in patients with less than six follicular oocytes compared with patients with six or more oocytes. Nonpregnant patients showed significantly higher incidence of apoptotic bodies in mural granulosa cell masses compared with pregnant patients. CONCLUSION(S) These results indicate that mural granulosa cell masses and cumulus cell masses may have different functions in follicular maturation. The incidence of apoptotic bodies in mural granulosa cell masses can be used as an indicator of success of IVF.

Aged Mouse Oocytes Fail to Readjust Intracellular Adenosine Triphosphates at Fertilization

Abstract Postovulatory aging of oocytes significantly affects embryonic development. Also, altered Ca2+ oscillation patterns can be observed in fertilized, aged mouse oocytes. Because Ca2+ oscillations depend on Ca2+ release and reuptake in the endoplasmic reticulum, and the latter relies on ATP availability, we simultaneously measured changes in intracellular ATP concentration ([ATP]i) and Ca2+ oscillations in fresh and aged mouse oocytes. We continuously assessed changes in [ATP]i from intracellular free Mg2+ concentration measured by fluorescent dye Magnesium Green (MgG) while intracellular Ca2+ concentration ([Ca2+]i) was monitored by Fura-PE3. At fertilization, MgG fluorescence was transiently increased concomitant with the first transient elevation in [Ca2+]i, indicating a relative decrease in [ATP]i. In fresh oocytes, it was quickly followed by a significant decrease below baseline, indicating a relative increase in [ATP]i. In contrast, in aged oocytes, such a decrease in MgG fluorescence was not observed. In a separate experiment, ATP content in fresh and aged oocytes was determined in vitro by the luciferin-luciferase assay. Intracellular ATP contents measured in vitro were comparable in unfertilized fresh and aged oocytes. Intracellular ATP content at 5 h after fertilization was increased in both oocytes, where fresh oocytes showed a significantly higher intracellular value than aged oocytes. These findings suggest that aged mouse oocytes fail to readjust the level of intracellular ATP at fertilization. Relative deficiencies of ATP at fertilization might lead to an altered Ca2+ oscillation pattern and poor developmental potency, which is commonly noted in aged oocytes.

Embryonic Heart Rates: Development in Early First Trimester and Clinical Evaluation

One hundred and forty-three women in the early first trimester of gestation were examined 364 times using transvaginal sonography, and the development of embryonic heart rate was studied. In each case gestational age was revised retrospectively by either recorded basal body temperature or ultrasound crown-rump length dating between 9 and 10 weeks. Embryonic cardiac activity could be detected as early as 37 days of gestation. In 133 continuing pregnancies, embryonic heart rate rose from an average of 97.7 beats per min at 36-38 days to 174.7 beats per min at 60-62 days. A significant correlation was seen between gestational age and embryonic heart rate (p less than 0.001). The regression equation for heart rate was as follows: heart rate = 3.850 x gestational age (days) -54.64 (r = 0.908, n = 347), in short, embryonic heart rate continued to rise about 4 beats per min every day until 8 weeks of gestation. In this series, 10 pregnancies resulted in spontaneous abortion in the first trimester, and all of them showed relative bradycardia. Embryonic heart rate measurements in 8 of them were below the 95% prediction intervals for normal heart rate plotted against gestational age. This study suggests that embryonic heart rate measurement by ultrasound may be a new method for dating early first trimester, and that first trimester bradycardia may be associated with a poor prognosis for the pregnancy.

Neonatal hyperbilirubinemia and the bilirubin uridine diphosphate‐glucuronosyltransferase gene: The common −3263T > G mutation of phenobarbital response enhancer module is not associated with the neonatal hyperbilirubinemia in Japanese

Abstract Background : Neonatal hyperbilirubinemia is frequent and severe in Japanese newborns. Previously, it has been reported that half of the Japanese neonates with severe hyperbilirubinemia carried the 211G > A (p.G71R) mutation of the bilirubin uridine diphosphate‐glucuronosyltransferase (UGT1A1) gene causing Gilbert syndrome. Recently, it was reported that the −3263T > G mutation in the phenobarbital response enhancer module in UGT1A1 was associated with the majority of cases of Gilbert syndrome. The gene frequency of the −3263T > G mutation was determined and the relation with neonatal hyperbilirubinemia in Japanese was studied.

Placental transfer of lidocaine hydrochloride after prolonged continuous maternal intravenous administration

We treated a patient with arrhythmia during pregnancy with prolonged intravenous administration of lidocaine hydrochloride. This was a case of twin-to-twin transfusion syndrome and the arrhythmia was caused by ritodrine therapy. In total, 14.1 g lidocaine (50 mg · hr−1 for 282 hr) were used. Since there are no descriptions of human placental transfer of lidocaine after such a prolonged continuous intravenous administration, we measured lidocaine concentrations in maternal and fetal serum, and in the amniotic fluid (AF) at delivery. Fetal serum lidocaine concentrations (donor: 0.83 μg · ml−1; recipient: 0.82 μg · ml−1) were lower than in the maternal serum (1.6 μg · ml−1), while the AF lidocaine concentrations (donor: 1.05 μg · ml−1; recipient: 1.04 μg · ml−1) were higher than those of the fetal sera. The fetal/maternal concentration ratios of lidocaine were 0.52 for the donor and 0.51 for the recipient, which were similar to those described previously after administration of lidocaine in labour.RésuméNous avons traité l’arythmie d’une parturiente porteuse de jumeaux avec de l’hydrochlorure lidocaline iv. Il s’agissait d’un syndrome de gémellité perfuseur-perfusé et l’arythmie était consécutive au traitement à la ritodrine. En tout, 14,1 g (50 mg · h−1 pour 282 h) de lidocaine ont été administrées. Comme on n’a jamais rapporté de données sur le transfert placentaire de la lidocaïne après une administration iv aussi prolongée, nous avons mesuré à l’accouchement les concentrations sériques maternelles et foetales de lidocaïne, et la concentration sérique de liquide amniotique (LA) de lidocaïne. Les concentrations sériques (perjuseur: 0,83 μg · ml−1; perfusé: 0,82 μg · ml−1) étaient moins élevées que dans le sérum matemel (1,6 μg · ml−1), alors que dans le LA (perfuseur: 1,05 μg · ml−1; perfusé; 1.04 μg · ml−1) les concentrations étaient plus élevées que dans le sérum foetal. Les rapports concentration foetus/ mère de lidocaïne étaient de 0,52 pour le perfuseur et de 0,51 pour le perfusé et identiques à ceux trouvés après l’administration de lidocaïne pendant le travail.

Pregnancy complicated by Werner's syndrome

Werner’s syndrome is defined as an autosomal recessive progeria. The majority of clinical symptoms are similar to those observed in elderly subjects and include wrinkling of the skin, greying of the hair, baldness, cataracts, diabetes type II, atherosclerosis and the other manifestations of ageing. Other features are commonly found in Werner’s syndrome which are unusual in normal ageing. These include laryngeal atrophy and tumours of connective tissue. The average life span in Werner’s syndrome is 47 years; cancer, myocardial infarction and cerebrovascular accidents are the main causes of death. This disease is associated with a mutation in the WRN gene, which codes for a helicase. Because hypogonadism is one of the common symptoms, pregnancy complicated by this syndrome is very rare and, to our knowledge, only two cases have been reported. We here report the characteristics of pregnancy in a woman with Werner’s syndrome. The woman was a 28 year old primipara, with a height of 147 cm and a weight of 41 kg before her pregnancy. Her menstrual cycles were regular. She had a bilateral cataract operation at the age of 26 and had been treated for a fatty liver, hyperlipidaemia, liver dysfunction and impaired glucose tolerance (Table 1). Her diabetes mellitus was treated by diet and her hyperlipidaemia with statins. She was referred to us at seven weeks of gestation with many unusual features. She had a short statue, a ‘bird’ face, grey hair, helomas, scleroatrophic skin and a high-pitched voice. A urinary hyaluronic acid test was positive, and X-rays showed osteosclerosis of the phalanges of her fingers. Because she had five of the cardinal signs and four of the other probable fulfilling signs of the diagnostic criteria, she was categorised as ‘probable’ for Werner’s syndrome. She had no obstetric problems such as pre-eclampsia, which occurred in a woman in a previous report. She stopped taking statins in early pregnancy. Some decrease in the level of liver enzymes was observed after mid-gestation. Serum total cholesterol and triglyceride increased after discontinuing the oral statins (Table 1). Although she had had impaired glucose tolerance before her pregnancy, she showed a normal profile during pregnancy. The fasting blood glucose and haemoglobin A1c levels were within normal ranges (Table 1), and the results of oral glucose tolerance tests at 7 and 30 weeks of gestation were normal. She was admitted to hospital at 30 weeks of gestation with a small antepartum haemorrhage and irregular uterine contractions. Her cervix was 1 cm dilated and was partly effaced. Ritodrine hydrochloride was given intravenously to suppress her premature contractions. At 34 weeks of gestation, progressive preterm labour occurred. Her infant daughter was born unaided and she cried at birth. She weighed 2.23 kg. She did not have any congenital abnormality. The woman did not lactate. Histological examination of the placenta showed an excess of syncytial knots, calcification and perivillous fibrin deposition (Fig. 1). After her delivery, the woman’s blood pressure and electrocardiogram were normal, while her hepatic enzymes were elevated (Table 1). Her baby has had no abnormality of growth at her last medical follow up, performed at one year and eleven months of age.

Sexual Difference in Early Fetal Crown-Rump Length versus Gestational Age in Pregnancies Arising from in vitro Fertilization

The purpose of this study was to investigate the influence of fetal sex on the crown-rump length (CRL) derived from pregnancies resulting from in vitro fertilization without the uncertain conception dating in the first trimester. With ultrasonography, we performed 334 and 353 examinations, on 88 male and 102 female fetuses, respectively. The relationship between gestational age and fetal CRL was explored with regression analysis: male CRL (mm) = 1.13 × gestational age (days) – 47.39 (r = 0.97), female CRL = 1.11 × gestational age – 46.44 (r = 0.97). These regression lines were mutually included in the 95% confidence intervals for each other. Our results indicate no statistically significant difference in the CRL between the two sexes, being supportive of using the identical fetal CRL criterion regardless of fetal sex for gestational age assessment with ultrasonography in the first trimester of pregnancy.

Development and Sexual Difference in Embryonic Heart Rates in Pregnancies Resulting from in vitro Fertilization

The purpose of this study was to investigate the gestational change of embryonic heart rates (EHRs) and to estimate the influence of embryonic sex on the EHR in pregnancies resulting from in vitro fertilization in the early first trimester. With transvaginal ultrasonography, we performed 92 and 105 examinations, on 27 male and 30 female embryos, respectively. The EHR increased gradually from 87 beats per min at 38 days of gestation to 189 beats per min at 62 days of gestation. The relationship between gestational age and EHR was explored by regression analysis: male EHR (beats per min) = 3.78 × gestational age (days) – 51.30 (r = 0.95), female EHR = 3.65 × gestational age – 44.56 (r = 0.95). These regression lines were mutually included in the 95% confidence intervals for each other. Our results indicate both a close positive correlation between the EHR and gestational age and no statistically significant difference in the EHR between male and female embryos. These findings suggest that the EHR measurement is a novel method for very early ultrasound dating with the identical EHR criterion without regard to embryonic sex.

Serological Study of -D-/D- Mother Free from Sensitization After Four Pregnancies: A Case Report

We present a case of a -D-/-D- mother free from isoimmunization after four pregnancies. Her parents denied consanguinity. Her parents, brother, and children are -D- heterozygous. -D- homozygote and heterozygotes in this family have more D antigen sites on their red blood cells than on those of the controls.