Matsuto Mochizuki

Effects of smoking on fetoplacental-maternal system during pregnancy

Fetoplacental function and maternal nutritional condition were assessed in order to clarify the mechanism of retarded fetal growth in pregnant women who smoked. Dehydroepiandrosterone sulfate (DHA-S) loading tests and measurements of cotinine, which is a major metabolite of nicotine, were also made. In heavy smokers, urinary estriol and serum levels of human placental lactogen (hPL) were lower than those in nonsmokers. There was no difference in maternal nutrition between smokers and nonsmokers. Heavy smokers demonstrated a lower conversion of DHA-S to estradiol than did nonsmokers. Levels of cotinine in maternal blood and umbilical cord blood of heavy smokers were remarkably higher than those in nonsmokers. Microscopic examination showed atrophic and hypovascular changes in placental villi from mothers who smoked. These results suggest that retarded fetal growth in heavy smokers is due to impairment of uteroplacental circulation as a result of the vasoconstricting effect of nicotine.

Tumor-associated antigen, TA-4, in the monitoring of the effects of therapy for squamous cell carcinoma of the uterine cervix. Serial Determinations and Tissue Localization

The level of tumor‐associated antigen (TA‐4) was determined in the serum and tumor tissue of patients with squamous cell carcinoma of the cervix by radioimmunoassay and immunoperoxidase techniques. Using an arbitrary limit of 2.5 ng/ml of serum, positive values were observed in 5.5% of healthy controls and 53.6% of patients with cervical squamous carcinoma. The mean value and incidence of elevation of serum TA‐4 increased significantly with advancing disease stage. There was, however, no significant increase in serum TA‐4 in the early stages of disease. Elevated TA‐4 in serum rapidly fell to normal within 72 hours after radical surgery, but remained elevated if complete excision could not be performed. In case of radiotherapy, TA‐4 levels in serum and tumor tissue often increased during the administration of the initial 2000 rad, and subsequently declined after the administration of a total of more than 4000 rad. The decline of serum TA‐4 to normal observed during radiotherapy was found to be closely correlated with the disappearance of viable cancer cells in histopathologic specimens from the cervix. Immunohistochemical TA‐4 staining was present in large cell nonkeratinizing carcinoma, but not in small cell nonkeratinizing carcinoma. These results indicate that the expression of TA‐4 antigen in cervical squamous carcinoma is related to differentiation of the tumor cells and that serum TA‐4 determination, despite its limitation for early diagnosis, provides a potential means for monitoring the effects of individual therapy for cervical squamous cell carcinoma.

Immunohistochemical localization of epidermal growth factor receptor and myc oncogene product in human placenta: implication for trophoblast proliferation and differentiation

Cytologic localization of epidermal growth factor receptor and myc oncogene protein product in developing human placenta was analyzed by avidin/biotin immunoperoxidase techniques with a monoclonal antibody to epidermal growth factor receptor and an affinity purified polyclonal antibody to myc protein product. Epidermal growth factor receptor was found to be almost exclusively localized to syncytiotrophoblast, paralleling the immunohistochemical localization of human chorionic gonadotropin. Since epidermal growth factor has been shown to stimulate human chorionic gonadotropin and human placental lactogen production by cultured early placental tissue, the present finding that epidermal growth factor receptor was localized to mitotically inactive syncytiotrophoblast suggests a role for epidermal growth factor receptor in the induction of differentiated function of trophoblast rather than trophoblast proliferation. By contrast, myc protein product was found to be predominantly localized to cytotrophoblastic cells, paralleling the autoradiographic distribution of replicating cytotrophoblast identified by tritiated thymidine labeling of placental explant. A close similarity between the cytologic localization of myc protein product and tritiated thymidine labeling of placental explant suggests that myc protein expression is linked to trophoblast proliferation. Furthermore, immunohistochemical cellular levels of both epidermal growth factor receptor and myc protein product were most pronounced in early placenta and declined in term placenta. Thus myc protein product and epidermal growth factor receptor seem to play a crucial role in the induction of trophoblast proliferation and differentiation, respectively, during the development of human placenta.

Pathophysiology and postnatal outcome of fetal hydrocephalus

At the National Kagawa Childrens or Kobe University Hospital, 24 cases of fetal hydrocephalus were managed between 1982 and 1988. There were 8 simple, 11 dysgenetic, and 5 secondary cases of hydrocephalus, and the fetal age at diagnosis ranged between 24 and 40 weeks of gestation (average 33.4 weeks). All were diagnosed using ultrasonography, with either magnetic resonance imaging or whole-body computed tomography, additionally performed in 10 patients to determine their usefulness in evaluating the morphology. Four patients underwent transabdominal or transvaginal cephalocentesis in the prenatal period and intracranial pressure was measured during the drainage of cerebrospinal fluid in two of these. Postnatal outcome was analyzed for each type of hydrocephalus. The results suggested that in such cases the fetal brain is subjected to extremely high intracranial pressures resulting from a mixture of hydrocephalic pressure and intermittent uterine constriction. Immediately after birth, the biparietal diameter was found to be increased by an average of 7.7 mm and the hydrocephalic state was transformed into the neonatal type characterized by macrocephaly and a relatively low intracranial pressure. Overall mortality was 25% and 16 of the 24 infants underwent the postnatal shunt procedure, largely at the neonatal stage. The follow-up period varied from 4 months to 6 years (average, 25.8 months for nonfatal cases) and the mean intelligence or developmental quotient was 45.2. There were no significant differences in postnatal outcome between the three major types of fetal hydrocephalus. Findings revealed that the length of the gestation period after the diagnosis of hydrocephalus has a significant effect on outcome (P<0.01). Based on these results, it is suggested that fetal hydrocephalus may be extremely hypertensive and that impairment of neuronal functional development accompanying its prenatal progression can be irreversible.

Examination of previous caesarean section scars by ultrasound

SummaryWe examined 84 lower segment caesarean section scars by ultrasonography near term. Seventy scars showed good healing with a thickness of the lower uterine segment of more than 3 mm; 14 scars showed poor healing with a thickness of less than 2 mm and loss of continuity. Among 70 patients with good healing, 24 patients delivered vaginally but the remaining 46 patients have had repeat caesarean sections for other obstetric indications. Intraoperative findings in these 46 patients were as follows: Grade I (no thinning of the lower uterine segment), 42; Grade II (thinning and loss of continuity of the lower uterine segment but fetal hair not visible), 4; Grade III (thinning of the lower uterine segment and fetal hair visible), 0. Fourteen patients with poor healing had repeat caesarean sections. Intraoperative findings in these 14 patients were as follows: Grade I, 0; Grade II, 9; Grade III, 5. These results indicate that ultrasound examination detect thinning of the lower uterine segment and may help to determine management.

Urinary Levels of Nitric Oxide Metabolites in Normal Pregnancy and Preeclampsia

Objective: The purpose of this study is to clarify the physiological role of nitric oxide (NO) in normal pregnancy and preeclampsia.

A novel change in cytologic localization of human chorionic gonadotropin and human placental lactogen in first-trimester placenta in the course of gestation

Cytologic localization of human chorionic gonadotropin and human placental lactogen in developing human early placenta was analyzed by avidin-biotin immunoperoxidase techniques with an affinity-purified polyclonal antibody to beta-human chorionic gonadotropin carboxyl terminal peptide and a polyclonal antibody to human placental lactogen. In 4- to 5-week placentas human chorionic gonadotropin and human placental lactogen were found to be primarily localized to cytotrophoblasts, whereas in 6- to 12-week placentas these substances were exclusively localized to syncytiotrophoblast. We previously reported that a similar change in cytologic localization of epidermal growth factor and its receptor from cytotrophoblasts to syncytiotrophoblast in first-trimester placenta appeared between 5 and 6 weeks of gestation. Because epidermal growth factor was demonstrated to stimulate human chorionic gonadotropin and human placental lactogen production by early placental tissues, their simultaneous expression, as well as epidermal growth factor and its receptor in the cytotrophoblast of 4- to 5-week placenta and in the syncytiotrophoblast of 6- to 12-week placenta, implies that human chorionic gonadotropin and human placental lactogen production by first-trimester placenta may be regulated in an autocrine manner, wherein epidermal growth factor may serve as the signal. These findings suggest that in very early placenta, before 6 weeks of gestation, no sequential expression of human chorionic gonadotropin and human placental lactogen closely linked to syncytia formation may exist and that both can be expressed in the cytotrophoblast or undifferentiated stem cell of villous trophoblast in very early placenta.

Biology of human trophoblast

In order to elucidate the regulation of placental growth, we have characterized the expression of proliferating cell nuclear antigen (PCNA), apoptotic DNA fragmentation and bc1‐2 protein in human placenta during pregnancy. PCNA and bc1‐2 protein expression were examined by immunohistochemical techniques, while the occurrence of apoptotic DNA fragmentation was assessed by in situ analysis of DNA 3′‐end labeling method. Both PCNA expression and apoptotic DNA fragmentation were found in cytotrophoblasts (C‐cells), being most abundant in early placenta, less abundant in midterm placenta and least abundant in term placenta. In contrast, bc1‐2 protein expression was found in syncytiotrophoblasts (S‐cells), being least abundant in early placenta, less abundant in midterm placenta and most abundant in term placenta. These data indicate that early placenta is characterized by the highly proliferative activity of C‐cells associated with the increased occurrence of apoptosis, whereas term placenta is characterized by the abundant expression of bc1‐2 protein in S‐cells.

Vascular reactivity in normal and abnormal gestation.

Preeclampsia is characterized by enhanced pressor responsiveness to angiotensin II. This report summarizes studies by our laboratory to investigate possible roles for calcium, sodium, membrane pumps, and the vasoactive hormones, atrial natriuretic peptide (hANP) and endothelin, in modulating the change in vascular reactivity characteristic of preeclampsia. Urinary calcium excretion, 1 alpha-25(OH)2D3 levels, and serum free calcium levels were all decreased, whereas parathyroid hormone levels and intraplatelet calcium concentrations were increased in women with preeclampsia. Erythrocyte sodium content was elevated, while red blood cell membrane Na-K-ATPase activity was decreased in patients with severe disease. Preeclamptics also had elevated levels of hANP, which failed to increase further when saline was infused or when blood pressure was increased transiently with angiotensin II administration. Finally, endothelin levels that are reduced in normal gestation, were increased in preeclampsia. While the cause of increased vascular reactivity is still unclear, there appear to be changes in the intracellular cation environment, combined with loss of compensating mechanisms, both at the membrane and humoral level, as well as enhanced concentrations of a potent vasoconstrictor in blood; all which lead to increases in vasoreactivity and blood pressure in preeclampsia.

Antigonadotropic Actions of GnRH Agonist on Ovarian Cells in vivo and in vitro

GnRH and its agonists have recently been shown to inhibit a variety of reproductive functions, in addition to its well-known gonadotropin releasing action in the pituitary. In order to determine the inhibitory mechanism and the site of action of these peptides in ovary, the direct actions of an agonistic analogue [D-Leu6, des-Gly-NH2] GnRH ethylamide on hypophysectomized immature rat ovaries in vivo and on rat luteal cells as well as porcine granulosa cells incubated in vitro were investigated. 125I-Labeled GnRH agonist, when injected to immature female rats, bound specifically not only to pituitary but also to ovaries. GnRH agonist inhibited hCG stimulation of progesterone production and ovarian weight augmentation in hypophysectomized immature female rats in vivo. FSH-stimulated induction of ovarian LH/hCG receptors and ovarian weight gain in diethylstilbestrol (DES)-treated hypophysectomized immature female rats were also suppressed by GnRH agonist. Moreover, treatment with GnRH agonist inhibited hCG-stimulated progesterone production by rat luteal cells incubated in vitro. In short time incubation of porcine granulosa cells obtained from medium follicles, the capacity and affinity of LH/hCG receptors were not affected by GnRH agonist. However, in long term culture of porcine granulosa cells obtained from small follicles, concomitant treatment with GnRH agonist markedly inhibited the induction of LH/hCG receptors stimulated by FSH and insulin. It may be possible that GnRH agonist prevents the differentiation of granulosa cells and subsequent acquisition of LH/hCG receptors, but has no effect on the LH/hCG receptors already induced. On the other hand, GnRH agonist delayed hCG-stimulated accumulation of cyclic AMP in porcine granulosa cells obtained from medium follicles. This delay of cyclic AMP accumulation may be responsible for the inhibition of progesterone production by ovarian cells. These findings suggest that GnRH agonist acts directly on ovarian cells and inhibits the action of FSH and LH/hCG, independently.