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We tested the ability of two separate nuclear reaction models, the binary cascade and JQMD (Jaeri version of Quantum Molecular Dynamics), to predict the dose distribution in carbon-ion radiotherapy. This was done by use of a realistic simulation of the experimental irradiation of a water target. Comparison with measurement shows that the binary cascade model does a good job reproducing the spread-out Bragg peak in depth-dose distributions in water irradiated with a 290 MeV/u (per nucleon) beam. However, it significantly overestimates the peak dose for a 400 MeV/u beam. JQMD underestimates the overall dose because of a tendency to break a nucleus into lower-Z fragments than does the binary cascade model. As far as shape of the dose distribution is concerned, JQMD shows fairly good agreement with measurement for both beam energies of 290 and 400 MeV/u, which favors JQMD over the binary cascade model for the calculation of the relative dose distribution in treatment planning.

Objective: To report the results of prenatal triple marker screening on a population of Japanese pregnant women. Methods: From April 1994 through March 1999, a total of 32,925 native Japanese women with singleton pregnancies requested a triple marker-screening test. Multiples of the median values for 3 markers and individual risks for each patient were calculated following adjustment for the Japanese weight correction factor. The risk cut-off values used for Down syndrome (T21), open spina bifida (OSB) and trisomy 18 (T18) were 1:295, 1:290, and 1:100, respectively. Follow-up information was collected postpartum and statistically analyzed. Results: Detection rates (DR) of T21 for women less than 35 years, over 35 years and overall were 58, 94, and 83%, respectively. DR of T18 for women less than 35 years, over 35 years and overall were 75, 79, and 79%, respectively. DR of open neural tube defects (ONTD) was 100%. Conclusions: The first cumulative data of an intervention program and prospective follow-up studies in Japan have proven to be similar to other published reports. Individual risk values were calculated for each pregnancy for T21, T18 and ONTD. This screening program is more effective than age-dependent screening for detecting T21, T18 and ONTD pregnancies.

Progesterone, prostaglandin and follicular fluid are reported to enhance the acrosome reaction through the influx of extracellular calcium into the cytoplasm of human spermatozoa. Prostaglandins are present within the male reproductive tract, and high concentrations of prostaglandins exist in seminal fluid. In order to investigate the mechanisms by which prostaglandins enhance the acrosome reaction through calcium influx, the intracellular calcium response induced by progesterone, prostaglandin E1 (PGE1), prostaglandin E2 (PGE2) and follicular fluid was measured using fura-2. PGE1 and PGE2 promoted calcium influx dose dependently through dihydropyridine insensitive calcium channels. Refractoriness of the elevation of intracellular Ca2+ concentration ([Ca2+]i) to a second stimulus occurred when 60 microg/ml PGE1 was administered 100 s after the prior administration of 60 microg/ml of PGE1, and similarly when 1 microg/ml of progesterone was administered 100 s after the prior administration of 1 microg/ml of progesterone. Refractoriness also occurred when 60 microg/ml PGE1 was administered after the prior addition of 60 microg/ml PGE2, but did not occur between PGE1 and progesterone. Pertussis toxin (PTX) did not modify the changes in [Ca2+]i after the addition of PGE1 or PGE2. In conclusion, PGE1 and PGE2 promoted calcium influx through PTX-insensitive calcium channels which appeared to be recognized by a common receptor different from that of progesterone.

CD44 is a cell-surface glycoprotein postulated to play a role in tumor-cell metastasis. We have examined the expression of the standard CD44 (CD44s), and alternative spliced variants of CD44 containing variant exons v6, v9, and v1O (CD44v6, CD44v9, and CD44v10 respectively) in 9 samples of normal cervix, 6 samples of cervical intraepithelial neoplasia (CIN), and 11 samples with invasive cervical carcinomas. RT/PCR demonstrated the presence of CD44s in all samples of normal cervix and those with invasive carcinomas. CD44v6 was also found in all normal cervical samples and in 9 tissue samples of invasive carcinomas. The results also suggested that some tumor specimens had several higher molecular transcripts containing exon v6 compared to specimens of normal cervix. Immunohistochemistry detected the presence of CD44s and the absence of CD44v10 in both epithelial and stromal cells in all specimens. In contrast, CD44v6 and CD44v9 were stained positive in epithelial cells but were absent in stromal cells. The intensity of CD44v6 and CD44v9 staining was strongest in normal cervical epithelium followed by CIN, invasive squamous cell carcinoma, and adenocarcinoma. In the malignant samples, heterogeneity in staining intensity among different clusters of tumor cells was observed. Furthermore, poorly differentiated and undifferentiated carcinomas from patients having poor prognosis did not stain at all. This study suggests that variant CD44 molecules may serve an important function in the cell contact of cervical epithelial cells, and that cervical epithelium acquires heterogeneity in the expression of CD44 adhesion molecules during carcinogenesis, which may be related to tumor metastasis.

This paper introduced newly developed computer-assisted learning materials and reports of a survey of junior college dental hygiene students who have used them.We authored new educational material to promote students' basic dental hygiene practice skills using a simulation software generator. A set of five developed materials were tested by 43 female second-year dental hygiene students during the second semester at a college in Chiba, Japan. The evaluation was conducted in the form of a questionnaire including open-ended questions. Students' opinions were analysed using characteristic diagrams, a troubleshooting tool that can be used to visually illustrate the causes and effects of a problem.The overall results of the evaluation were positive. The students were given five sets of simulation learning materials (SLMs). Eighty-three percent of the students felt that they could carry out independent study of clinical practice better after the virtual practice. Ninety-three percent of them felt that the exercises should be continued in the future, and eighty-eight percent of them felt that this virtual practice deepened their interest in other classes and training sessions. All of the students found the virtual practice beneficial for their learning.The present results suggest that the students became conscious of their lack of knowledge through SLMs. These findings indicate that SLMs for practicing basic clinical procedures is beneficial.

To elucidate the possible regulation of endometrial growth, we investigated the expression of tenascin, an extracellular matrix protein associated with cell proliferation and morphogenetic events, in relation to the mitotic marker ki-67 antigen in human endometrium throughout the menstrual cycle. The endometrial tissue was obtained from women with regular menstruation undergoing hysterectomy due to benign uterine disease. Cryostat sections were subjected to double immunohistochemistry to detect both tenascin and ki-67. During the proliferative phase, an intense deposition of tenascin fibrils occurred in the extracellular matrix of the stroma immediately subjacent to the uterine epithelium that showed the highest proportion of ki-67 positive cells. In the secretory phase, the amount of tenascin expression and the proliferative index of the epithelium were markedly reduced. The topological immunoreactivity of tenascin, however, was clearly related to ki-67 localization close to the tortuous endometrial gland and spiral artery. Expression of tenascin, showing changes depending on the menstrual phase, appears to be influenced by ovarian steroid hormones, and it might play a role in regulating human endometrial development by interaction with mitotic cells.

The purpose of this study was to evaluate the efficacy of ipriflavone in preventing bone loss, decreasing in serum cholesterol and decreasing the rate of appearance of vasomotor symptoms, as well as the effects of ipriflavone on reduction of myoma volume by estrogen deficiency during treatment with the GnRH analog leuprolide. One hundred two women (mean age, 44.3 +/- 0.53 yr) receiving leuprolide therapy for uterine leiomyoma were randomly allocated to two groups (group A, leuprolide only; group B, leuprolide with ipriflavone). Bone mineral density of the lumbar spine was measured by dual-energy x-ray absorptiometry before and after treatment for 6 months. Levels of serum total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol, and low-density lipoprotein cholesterol (LDL-C) were measured before treatment and after 3 and 6 months of treatment. Subjects were asked to report the appearance of vasomotor symptoms throughout treatment. Myoma node volumes were measured before treatment and after treatment for 6 months. Bone mineral density was reduced in both groups, with reduction rates of -5.26% in group A and -3.70% in group B (P0.01 vs. group A). Changes in bone markers were not significant in either group. TC was significantly increased in both groups, and TG levels were increased significantly after 3 and 6 months of treatment in group A but not in group B. There was no significant difference between these two groups in amount of increase of either TC or TG. LDL-C levels were increased significantly after 3 and 6 months of treatment in both groups, and the differences between the groups (11.7% in group A vs. 7.5% in group B at 3 month and 22.6% in group A vs. 8.4% in group B at 6 month) were significant. Severe vasomotor symptoms were reduced in group B. The rates of reduction of myoma volume were 49.8% in group A and 52.9% in group B; this difference between groups was not significant. Ipriflavone efficaciously alleviated the adverse effects of estrogen deficiency such as bone loss and increase in LDL-C level, and the ability of leuprolide therapy to reduce myoma volume was not decreased by ipriflavone administration.

Elongin A is the transcriptionally active subunit of the Elongin complex, which strongly increases the rate of elongation by RNA polymerase II by suppressing the transient pausing of the polymerase at many sites within transcription units. In the present study, we obtained the cDNA sequence of the mouse Elongin A gene (Tceb3) and characterized its genomic structure. The deduced 773-amino acid sequence of mouse Elongin A shows 91% and 81% identity with rat and human Elongin A, respectively. The Elongin A gene was mapped to mouse chromosome 4D3 by fluorescence in situ hybridization.

To confirm the characteristics of the stromal cells of Wharton's jelly, we investigated the morphological changes in these cells during each trimester of pregnancy. We evaluated the cytoskeletal features of these cells by examining immunohistochemically the localization of one of the contractile proteins, alpha-smooth muscle actin (ASMA). After the second trimester, the stromal cells of Wharton's jelly were stained with ASMA antibody, exhibited the ultrastructural characteristics of the myofibroblasts, and began to express numerous microfilaments in the cytoplasm. Postembedding immunogold labeling detected immunoreactivity for ASMA on these microfilaments. The finding indicated that the stromal cells of Wharton's jelly undergo a time-dependent maturation involving the differentiation of myofibroblasts during the last 6 months of pregnancy. These cells possess a contractile function that may help to protect the umbilical vessels from compression, considering that ASMA was detected in the microfilamentous bundles.

The hormonal patterns during menstrual cycle, which consist of cyclic alterations in gonadotropins, estradiol, and progesterone, are controlled by hypothalamic-pituitary-ovarian feedback mechanism. GnRH produced in hypothalamus acts on the pituitary cells to secrete FSH and LH, which stimulate the follicular development. The developed follicles secrete estradiol, progesterone, inhibin, activin, and follistatin. Estradiol and progesterone, at different concentrations and/or ratios, either positively or negatively control the feedback of hypothalamic-pituitary axis in regulating the secretion of GnRH, FSH and LH. Inhibin and follistatin selectively suppress, whereas activin enhances the secretion of FSH in the pituitary. Recently, various additional factors produced by the ovary have been identified to contribute to the follicular development by paracrine and/or autocrine regulation as well as to feedback on hypothalamic-pituitary unit.

This study was undertaken to investigate the influence of past pregnancy and delivery on the perimenopausal status in 144 women (48-52 years of age). Individual interviews to ascertain the personal profile, medical history, events at pregnancy, delivery and puerperium, and the kind and severity of climacteric symptoms were held. The subjects were divided into two groups by means of a simplified menopausal index (SMI): group A had climacteric symptoms (SMI50) and those in group B had mild symptoms (SMIor = 50). The analysis of the principal components, and the correlation among the variables revealed that three factors--the impression and events at past pregnancies and deliveries, reproductive functions, and the psycho-character properties--had a significant influence on the status at the perimenopause, but the social-environmental factors appeared to have no significant influence on the severity of climacteric symptoms.

The effects of endothelin-1 (ET-1) on luteinized human granulosa cells (L-HGCs) have not been examined. It is well known that there are differences of actions of several autocrine/paracrine regulators between L-HGCs and GCs of other species, and therefore the present study was designed to examine the effects of ET-1 1) on intracellular Ca2+ concentrations ([Ca2+]i) using the Ca(2+)-responsive fluorescent indicator Fura-2, 2) on cell proliferation by the nonradioactive method using bromodeoxyuridine, and 3) on basal and gonadotropin-stimulated steroidogenesis, and to examine the expression of ET receptor messenger RNA (mRNA) using freshly isolated and cultured L-HGCs obtained from patients undergoing in vitro fertilization. ET-1 increased [Ca2+]i in L-HGCs in a dose-dependent manner between 1 and 1000 nmol/L. High concentrations (100-1000 nmol/L) of ET-1 produced a more rapid and transient increase in [Ca2+]i than that observed with low concentrations (1-10 nmol/L) of ET-1. The increase in [Ca2+]i elicited by ET-3 (1000 nmol/L) and IRL-1620 (1000 nmol/L), a selective ETB receptor agonist, was 16% and 3% (vs. ET-1, 100%), respectively. BQ-123 (1000 nmol/L), an ETA receptor antagonist, inhibited the increase in [Ca2+]i elicited by ET-1 (by 50% at 1000 nmol/L ET-1 and by90% at500 nmol/L ET-1). mRNAs for the two known receptor subtypes (ETA and ETB) were also present in L-HGCs; however, the expression of ETA receptor mRNA was much greater than that of ETB receptors. ET-1 stimulated cell proliferation in L-HGCs in a dose-dependent manner (1000 nmol/L, 210.5 +/- 13.1%; 100 nmol/L, 198 +/- 11%; 10 nmol/L, 146 +/- 18%; and 1 nmol/L, 103 +/- 9%; vs. control, 100%). These stimulatory effects were completely blocked by BQ-123 (1000 nmol/L). ET-3 and IRL-1620 had no effects on cell proliferation in L-HGCs. Significant stimulatory effects on cell proliferation by the calcium ionophore, ionomycin (10-1000 nmol/L), were observed. ET-1, ET-3, and IRL-1620 attenuated basal progesterone secretion in L-HGCs. These results suggest that ETA receptor predominantly exist in L-HGCs and that ET-1 may stimulate cell proliferation of L-HGCs by increasing [Ca2+]i via ETA receptors.

Germline mutations in the von Hippel-Lindau tumor suppressor gene (VHL) predispose individuals to a variety of tumors, including renal carcinoma, hemangioblastoma of the central nervous system, and pheochromocytoma. Here, a cellular transcription factor, Elongin (SIII), is identified as a functional target of the VHL protein. Elongin (SIII) is a heterotrimer consisting of a transcriptionally active subunit (A) and two regulatory subunits (B and C) that activate transcription elongation by RNA polymerase II. The VHL protein was shown to bind tightly and specifically to the Elongin B and C subunits and to inhibit Elongin (SIII) transcriptional activity in vitro. These findings reveal a potentially important transcriptional regulatory network in which the VHL protein may play a key role.

Mammalian transcription factor IIF (TFIIF) is a heterodimer composed of approximately 30-kDa (RAP30) and approximately 70-kDa (RAP74) subunits. TFIIF has been shown to bind RNA polymerase II and control the activity of the enzyme in both the initiation and elongation stages of transcription. Although previous studies have established a role for RAP30 in assembly of the preinitiation complex and in transcription initiation, information on the function of RAP74 in these processes has been lacking. Using a highly purified transcription system and assays that permit sensitive measurement of the contributions of both RAP30 and RAP74 to TFIIF function, we have investigated the roles of these TFIIF subunits in transcription initiation and elongation. Results of template competition experiments indicate that both RAP30 and RAP74 contribute to the formation of stable preinitiation intermediates containing RNA polymerase II. Investigation of the role of TFIIF in transcription initiation indicates that both RAP30 and RAP74 function in synthesis of the first few phosphodiester bonds of nascent transcripts and in formation of Sarkosyl-resistant pre-initiation intermediates. Finally, kinetic experiments indicate that both RAP30 and RAP74 function in TFIIF-mediated stimulation of the rate of RNA chain elongation by RNA polymerase II.

The purpose of this study was to investigate the combined effect of bromocriptine (Brc) and clomiphene citrate (Cl) treatment on 40 patients with normoprolactinemic amenorrhea who failed to respond to Cl alone. The ovulation rate in this treatment was 57.3% (23/40) in the 40 cases, 55.6% (99/178) in 178 cycles; the pregnancy rate was 26.7%. This treatment was effective in 14 of 21 women with polycystic ovary-like syndrome (66.7%). Among those women who responded to treatment, prolactin (PRL) and LH levels were significantly decreased. Estradiol and progesterone levels were significantly increased in the patients who responded. Before treatment, the responsiveness of LH to LHRH among responders to Brc/Cl therapy was significantly higher than among the nonresponders. After treatment, the LH-releasing response following a conjugated estrogen injection in the patients who responded to the treatment was significantly greater than that in the patients who did not respond. The results suggest that the therapeutic effect of this treatment may be primarily due to the restoration and improvement of the impaired hypothalamo-pituitary axis.

From January 1981 to December 1990, intracardiac repair of tetralogy of Fallot in 148 pediatric patients, with one surgical death, was directed toward preservation of the native pulmonary valve. Using the accepted preoperative angiographic criterion for the pulmonary valve annulus area (PVA) of 1.8 cm2/m2, 85 patients were candidates for transannular right ventricular outflow patch (TAP). However, in 54 patients with a mean PVA of 1.5 cm2/m2 (range 1.06-1.76), the valve was preserved without using TAP because the morphological changes (cusp thickening and annular distensibility) seemed acceptable for preservation in view of its probable hemodynamic efficiency and growth potential. A morphological classification of pulmonary valve changes has evolved. Retrospectively, 24 (77%) of the 31 patients with TAP had moderate to severe cusp thickening and ring rigidity; this incidence was significantly higher (p0.001) than that in preserved patients (18 of 54 or 33%). The incidence of morphological changes increased with operative age; that is, 2 of 13 (15%) patients younger than 1 year versus 23 of 40 (58%) patients older than 4 years (p0.01). All 54 patients with preserved pulmonary valves were catheterized one month postoperatively. The intraoperative right to left ventricular systolic pressure ratio (RVP/LVP) decreased significantly (p0.005) in one month, from a mean of 0.79 (range 0.44-1.36) to 0.57 (range 0.36-0.97). The PVA increased from a mean of 1.5 to 1.9 cm2/m2 (range 1.20-2.65), and the rate of its increase was significantly larger (p0.005) as operative age decreased. Pulmonary valve regurgitation of greater than mild degree occurred in 8 of 54 (15%) patients with the valve preserved.(ABSTRACT TRUNCATED AT 250 WORDS)

It has been reported that insulin-like growth factor (IGF) may play an important role in the feto-placental environment during pregnancy. The present study was undertaken to investigate the effect of IGF-I and IGF-II on prolactin (PRL) release from human decidual cells in vitro and in vivo. The human decidua in early pregnancy was obtained by DC, and was enzymatically dispersed into a monocellular suspension. Monolayer cultures of these cells were exposed for 96 hours to either control media or medium supplemented with IGF-I and IGF-II. PRL levels in the media were measured by EIA. Five days after dispersion, the intracellular calcium concentration [( Ca]2+i) in cultured decidual cells was measured by the Fura-2 fluorescence method with a Spectrofluorometer. PRL and IGF-I levels in amniotic fluid of 2nd trimester and term pregnancy were measured by RIA for in vivo study. In an in vitro study, IGF-I increased PRL release from decidual cells significantly during a 96 hour culture. However, IGF-II did not enhance this PRL release. IGF-I stimulation had no effect on [Ca]2+i. In an in vivo study, amniotic fluid IGF-I levels in the 2nd trimester (139.8 +/- 28.1 ng/ml) were significantly higher than those in term pregnancy (46.4 +/- 14.2 ng/ml), and a significant positive correlation (r = 0.852, p less than 0.001) was observed between IGF-I and PRL levels in the amniotic fluid. The present in vivo and in vitro studies indicated that IGF-I plays an important role in PRL release from decidua into amniotic fluid.(ABSTRACT TRUNCATED AT 250 WORDS)

In order to elucidate the possible involvement of endothelin-1 (ET-1) in the development of the feto-placental unit, plasma concentrations of immunoreactive ET-1 during pregnancy and delivery were measured in the current study. Forty-four pregnant women (8 in the 1st trimester, 8 in the 2nd trimester, 12 in the 3rd trimester and 16 at delivery), 6 at 1 month postpartum and 14 non-pregnant women volunteered to participate in this study. Plasma levels of immunoreactive ET-1 in all subjects were measured by specific radioimmunoassay. Plasma cortisol, prolactin and oxytocin levels during delivery were also measured by RIA. Although plasma ET-1 levels did not change during the 1st (1.3 +/- 0.4 pg/ml, mean +/- S.E.M., n = 8) or 2nd (1.7 +/- 0.3 pg/ml, n = 8) trimester, they were significantly (p less than 0.05) higher in the 3rd trimester (2.6 +/- 0.3 pg/ml, n = 12) than those of the non-pregnant controls (1.7 +/- 0.3 pg/ml, n = 14) and returned to the control levels within 1 month post-partum (1.8 +/- 0.1 pg/ml, n = 6). Maternal ET-1 levels before labor onset (2.3 +/- 0.4 pg/ml, n = 16), at delivery (3.0 +/- 0.7 pg/ml) and 24 hours post-partum (2.7 +/- 0.5 pg/ml) showed no significant differences among them. On the other hand, ET-1 levels in cord plasma (7.0 +/- 1.2 pg/ml) were significantly higher (p less than 0.05) than those in maternal plasma. Furthermore, there were no significant correlations between plasma ET-1 levels and those of other hormones (cortisol, prolactin and oxytocin) during delivery. From these results, we concluded that maternal plasma ET-1 levels significantly increased in the last trimester and returned to non-pregnant levels within one month post-partum.

We investigated the gene expression of the alpha chain of C4b-binding protein (C4bp alpha) in a variety of tissues, and in liver cell and hepatoma lines. C4bp alpha mRNA was detected in the liver, but not in the other tissues examined. The constitutive gene expression of C4bp alpha by a hepatoma line, HepG2, was significantly augmented by treatment with monocyte-conditioned medium (MoCM), 12-O-tetradecanoylphorbol-13-acetate (TPA), interleukin-6 (IL6) and tumor necrosis factor (TNF) but not by a calcium ionophore (A23187) or interleukin-1 beta (IL1 beta).

The floor of the oral cavity and mandibular of a 27-year-old female was reconstructed with the D-P flap after resection of a giant keloid in the submandibular region and a radiation scar of the mandibular. The mandibular was reconstructed with a transplantation graft of the iliac bone and the functional properties and appearance were good.

Fifteen patients (8 male and 7 female) with multiple myeloma, who were admitted to our hospital between July 1986 and August 1988 and suffering from pain and hypercalcemia, were treated with synthetic calcitonin derivative (elcatonin: ECT). ECT was administered intravenously at a dose of 10-640 units twice daily. Seven patients were treated with ECT (ECT group), and eight patients received combination treatment with ECT and other form of chemotherapy (combination group). With regard to the pain score (PS), significant analgesic effects in both groups were observed during 1-4 week treatments (p less than 0.05). There were no significant differences in PS between two groups. Serum calcium levels in the combination group at 1 and 4 weeks were significantly lower than the initial value (p less than 0.05). Hypocalcemia was not seen in any of the patients. Urinary excretion of calcium at 1 week in ECT group was higher than the initial value (p less than 0.05). The observed toxicities of ECT were slight nausea and vomiting in only 2 patients. These findings suggest that ECT is an useful agent for the treatment of pain and hypercalcemia accompanied with multiple myeloma.

Eleven unconjugated steroids were measured daily during a complete cycle in the peripheral plasma of 6 normally menstruating baboons (Papio hamadryas) by means of a radioimmunoassay procedure and the levels were compared with those found previously in 17 normally menstruating women. The patterns of progesterone and 20alpha-dihydroprogesterone were very similar to those found in women throughout the entire menstrual cycle. However, the ratio of these steroids differed markedly from that found in women. A great similarity of the follicular phase and the peri-ovulatory period profiles was observed in both species for 17-hydroxy-progesterone, pregnenolone, androstenedione, oestradiol and oestrone. The pattern of oestradiol:oestrone ratios was similar in both species. However, an elevation of the above five steroids, typical for the human luteal phase, was not found in baboons. The increase of testosterone values, seen in women at mid-cycle, was not detected in baboons. The plasma concentrations were lower in baboons than in humans for all the above steroids.

SUMMARY A rapid 5 h radioimmunoassay method for the determination of progesterone and oestradiol in the plasma of non-pregnant women is described. Due to the high specificity of the antisera used, it is possible to perform the radioimmunoassay directly on the ether extracts of plasma, without employing chromatographic purification of the steroids. Evidence is presented indicating that the rapid assay is almost as reliable as the previously described radioimmunoassay method which involves chromatography. The within-assay and between-assay coefficients of variation in the progesterone assay were 7.74 and 14.9 and in the oestradiol assay 7.36 and 18.1 respectively. Comparisons between increasing doses of authentic hormone and endogenous hormone extracted from plasma indicated no deviation from parallelism. Progesterone and oestradiol were assayed in 300 plasma samples by the rapid method and by the method involving chromatography. The slopes obtained by a regression analysis were close to unity for both progesterone and oestradiol (1.04 and 1.06, respectively), the y-intercepts were −0.21 and −0.16 and the correlation coefficients 0.98 and 0.88, respectively. When the data obtained by both techniques in fourteen menstrual cycles were compared, the results were practically identical. In ten repeated studies conducted by four investigators it was shown that two workers can complete the assay of both progesterone and oestradiol in twentyfive plasma samples in duplicates within 5 h. The same time is required for the assay of either progesterone or oestradiol in twenty-five duplicates by one worker.

In an attempt to find suitable animal models to aid in the study of the reproductive processes of the human male, plasma levels of unconjugated pregnenolone, 17-hydroxypregnenolone, dehydroepiandrosterone, 20α-dihydroprogesterone, 17-hydroxyprogesterone, androstenedione, testosterone, dihydrotestosterone, oestrone and oestradiol were measured in 18 male baboons and 10 male rhesus monkeys and the steroid levels were compared with those previously established in normospermic, middle-aged men. Significant species differences were found with regard to the three Δ5-steroids studied; whereas the approximate relationship of pregnenolone to 17-hydroxypregnenolone and dehydroepiandrosterone was 1:2:4 in men, the corresponding relationship was 1:5:30 in rhesus monkeys and 1:10:10 in baboons. Similar levels of 20α-dihydroprogesterone were found in the three species. On the other hand, the 17-hydroxyprogesterone levels in baboons were much lower and the levels of androstenedione lower than those found in men and in rhesus monkeys. No species difference was found with regard to circulating testosterone levels. However, both rhesus monkeys and baboons exhibited much higher levels of dihydrostestosterone than did men. Oestrone levels were higher in baboons than in men and oestradiol levels were higher in rhesus monkeys than in men and in baboons. The significant differences in circulating steroid levels suggest that further studies (including i.a. steroid analyses in testicular tissue, seminal plasma and spermatic artery and vein following both stimulation and suppression of testicular endocrine function) are required before preference can be given to any of the two species studied as a suitable animal model for the study of new fertility regulating agents.

Two consecutive pregnancies were induced in three hyperprolactinemic primary infertile patients bearing intrasellar microadenoma without visual disturbance. Pretreatment prolactin levels, 224 -496ng/ml, were normalized by 10-22.5mg/day of bromocriptine and the initial conceptions occurred. No abnormal sign due to pituitary enlargement was detected throughout the period of gestation and they delivered single full-term babies. Since no significant change was observed in puerperium, it was decided to give the second bromocriptine therapy. The prolactin levels, 80-190ng/ml, measured before the second therapy were apparently lower than those seen prior to the first therapy and the minimum effective dose used to induce the second pregnancy (5-7.5mg/day) was identical to 1/3-1/2 of the first one. The course of the second pregnancy of each case was also uneventful and the babies were all normal and mature. The magnitude of the rise in the prolactin level during the second pregnancy was smaller than that observed during the first pregnancy. The present longitudinal study throughout two consecutive pregnancies in three cases indicates that resolution of prolactinoma was resulted in some extent after withdrawal of stimulative factors associated with pregnancy. Hence, hyperprolactinemic infertile cases with intrasellar microadenoma can be subjected to bromocriptine therapies inducing consecutive pregnancies under intensive follow-up.

Morphological and endocrinological studies were performed on a 19-year-old case of an arrhenoblastoma with marked virilization. The tumor was an intermediate type of Meyer's classification. Histochemically, 3 beta-HSD and G-6-PDH activities were demonstrated in Leydig cells. These cells also had ultrastructures typical of steroid-producing cells. Basal blood cells of pregnenolone, dehydroepiandrosterone, testosterone (T), androstenedione (A), progesterone, estradiol, estrone, LH, and FSH were determined pre- and postoperatively. T and A showed a very high level preoperatively, and were markedly decreased immediately after removal of the tumor. Stimulation of tumor cells by HMG-HCG did not show any significant changes in their main products of T and A. These findings suggest that Leydig cells of the present tumor produced mainly A and T independently on gonadotropins, and these hormones had virilized the patient.

In an attempt to analyze the multiple changes and interactions in circulating steroid levels in the peri-ovulatory and peri-menstrual periods, the plasma levels of immunoreactive luteinizing hormone (LH), progesterone and unconjugated pregnenolone, dehydroepiandrosterone, testosterone, oestradiol and oestrone were assayed daily during a complete cycle in 17 normally menstruating women. In 14 of the 17 subjects studied androstenedione and unconjugated dihydrotestosterone were also estimated. The day of the LH-peak and the first day of menstruation, respectively, were used to synchronize the peri-ovulatory and peri-menstrual plasma levels of the various steroids. With the exception of dehydroepiandrosterone and dihydrotestosterone, the plasma levels of all steroids exhibited significant, but different changes during the cycle. Testosterone levels showed a slight but significant increase around the LH-peak, whereas the levels of pregnenolone and androstenedione were higher in the post-ovulatory than in the pre-ovulation periods. The levels of oestradiol and oestrone, as well as the ratios of oestradiol to oestrone gradually increased from the low values observed in the early proliferative phase to pre-ovulatory peak values. The relationship between peaks of oestradiol and oestrone and that of LH exhibited great individual variation. The same was true for the individual oestradiol to oestrone ratios. The combination of several steroidal signals did not improve the predictive value of the analyses. However, an increase of individual progesterone values by at least 0.35 ng/ml from the day preceding the LH-peak to the day of the LH-peak was observed in 13 of the 17 subjects. It is suggested that for the early detection of the LH surge and prediction of the subsequent ovulation daily assays of plasma progesterone are of more value than the assay of the other steroids investigated.

A case of precocious puberty due to a hypothalamic hamartoma is presented. Concentrations of plasma LH and FSH, testosterone and its derivatives were found to be elevated. Circadian rhythms of LH were also observed. After removal of the mass, plasma LH and FHS concentrations declined to nearly half the preoperative levels.

Plasma levels of unconjugated oestrone, oestradiol, oestriol and progesterone were serially studied in six uncomplicated patients in the mid-trimester of pregnancy before and during abortion induced by purely mechanical stretching of the uterus by laminaria and rubber balloon. Variability of these hormonal levels among patients was significant before the treatment. In four cases where the fetus was aborted alive, the plasma value of these hormones remained at a high level during the treatment with the exception of oestriol level in one case. In two early mid-trimester cases the fetus died during the treatment and plasma oestriol dropped significantly, while the level of the other hormones remained raised until fetal delivery. In these two cases the apparent onset of labour was noted before fetal death. It was concluded that the onset and progress of labour by mechanical stretching of the uterus is probably unrelated to the steroid hormones estimated in the present study.

Human trophoblast and endometrial cell interaction was studied by using tissue culture techniques as an in vitro model experiment on ovum implantation. Mixed cultures of trophoblast of the early gestational stage and endometrial cells of the mid-secretory phase were performed, and the interactions between these cells were observed by using a phase contrast microscope and microcinematographs. Trophoblast cells showed marked affinity and firm attachment to endometrial cells, and their outgrowth and maintenance were significantly better in the mixed cultures with endometrial cells than in the cultures of trophoblast alone. These characteristic interactions appeared to be closely related to attachment and anchoring of trophoblast to endometrium, and suggested the supportive and nutritive roles of endometrial cells in trophoblast development in the process of implantation. Trophoblast cells showed active cell locomotion, pinocytosis, and phagocytosis. Aggressive action on epithelial endometrial cells observed in the cinematographs seemed to be suggestive of the cytolytic activity of trophoblast cells. Trophoblast cells invaded endometrial cell sheet in the same manner as malignant tumor cells in vivo. Outgrowth and invasion of trophoblast cells were finally arrested by transformed endometrial cells similar to decidual cells morphologically. These results demonstrated the in vitro decidualization of endometrial cells and suggested their control action against trophoblast invasion.

Of 20 female and 14 male patients with chromophobe adenomas, 4 male patients, aged 38 to 47 years, and one female patient, aged 54, showed above normal basal plasma levels of follicle-stimulating hormone (FSH). An in vitro tissue culture study that was performed using chromophobe cells obtained from 3 of the 4 male patients and the 1 female patient showed moderate FSH secretion into the culture media. The secretion of FSH by pituitary chromophobe adenomas might be more frequent than hitherto suspected.

Effects of open-heart surgery on renal function were studied in 38 patients who had uneventful postoperative course with no sign of acute renal failure. Of these, 19 underwent aorto-coronary bypass grafting (Group 1) and 19 underwent valve replacement (Group 2). During cardiopulmonary bypass (CPB), renal plasma flow (RPF) and glomerular filtration rate (GFR) significantly decreased in both groups as compared to the preoperative values. The decreased GFR remained significantly low on the day of operation in both groups and returned to the preoperative level on the 1st postoperative day (POD) in group 1 and on the 3rd POD in group 2. On the other hand, plasma antidiuretic hormone (ADH) level markedly increased during CPB and remained significantly higher than the preoperative level through the 1st POD. Despite the decreased GFR and increased ADH, however, urinary flow markedly increased during CPB in both groups and remained at significantly higher level than the preoperative values through the 3rd POD in group 1 and through the 5th POD in group 2. Fractional excretion of sodium significantly increased during CPB in both groups and remained high through the 1st POD in group 1 and through the 3rd POD in group 2. These data demonstrate that the renal function of urine concentration is disturbed in the early phase following open-heart surgery even in the patients who have uneventful postoperative course.

We report fatal transfusion-associated graft-versus-host disease (GVHD) in a patient who was not severely immunosuppressed. A 58-year-old man received 800 ml of fresh whole blood from his son and an unrelated volunteer donor during open heart surgery. On the 10th day after the operation, he suddenly had a high fever, followed by generalized skin rash and liver dysfunction. Pancytopenia due to bone marrow aplasia developed a week later. A skin biopsy revealed a cutaneous lesion highly compatible with acute GVHD. The patient did not respond to high-dose methylprednisolone therapy, and died of multiple organ failure on the 18th day after the operation.