Search

The serum concentration of leptin in 10 week old virgin ICR mice assessed by RIA was 1.70 ± 0.08 ng/ml. The serum leptin concentration in the pregnant mice mated at 10 weeks of age significantly increased from day 11 of pregnancy and reached a peak on day 17 of pregnancy (42.2 ± 4.8 ng/ml). After the delivery, the serum leptin concentration rapidly decreased and reached the level of the virgin mouse on the seventh day in the puerperium. Tissue contents of leptin in the placenta, the decidua, the uterus, and the adipose tissue were between 40 to 130 ng/g wet tissue. However, leptin mRNA was expressed only in the adipose tissue and the level of leptin mRNA on days 13 and 17 of pregnancy increased 3- to 5-fold compared with that of virgin mouse. Tissue content of leptin in the adipose tissue significantly increased from day 17 of pregnancy compared with that of the virgin mouse. The m-leptin secretion from the adipose tissue also significantly increasedin vitro.These results suggest that leptin, which was secreted by adipose tissue, may play important roles in mouse reproduction after midpregnancy.

Parathyroid hormone-related peptide (PTHrP) is found in very high concentrations in the milk of various mammals. However, little is known about its physiological role in this fluid. To obtain detailed profiles of PTHrP in milk, we measured the concentrations of PTHrP in human milk by two different region-specific assays, PTHrP(1–87) (N-PTHrP) and PTHrP(109–141) (C-PTHrP). We also examined the correlations between PTHrP and Ca concentrations in milk as well as the correlations between PTHrP and secreted milk volume. The levels of N-PTHrP and C-PTHrP were relatively low after delivery and gradually increased to 13·87 ± 2·40 nmol/l (mean ± s.e.m.) and 56·39 ± 11·31 nmol/l respectively on the 10th day postpartum. N-PTHrP concentration remained steady until the 6th month postpartum when weaning starts, at which point it decreased slightly. C-PTHrP levels changed in a similar way to N-PTHrP levels but were 2- to 5-fold higher. Milk Ca concentration, and content, correlated with C-PTHrP concentration, and content (r=0·422 and r=0·769 respectively; Pr=0·524, P These results suggest that PTHrP in human milk may play some role in the maintenance of lactation through the N-terminal region and in promoting Ca transfer into milk via the C-terminal region. Journal of Endocrinology (1997) 153, 445–451

OBJECTIVE We examined the effects of transforming growth factor-beta 1 (TGF-beta 1) on intracellular parathyroid hormone-related peptide (PTHrP) production in a human ovarian cancer cell line (HOC-21). METHODS Various concentrations of TGF-beta 1 were added to a culture medium, and the numbers of cells were counted. HOC-21 cells were cultured with or without TGF-beta 1, and intracellular PTHrP was measured by a radioimmunoassay kit that recognized the carboxy-terminal protein while counting the number of cells. Furthermore, a 125I-TGF-beta 1 binding assay was carried out. Values were analyzed statistically using an analysis of variance followed by an unpaired t-test. RESULTS Five ng/ml of TGF-beta 1 inhibited the cell growth on Day 1 after plating. The TGF-beta 1 significantly (p < 0.05) stimulated intracellular PTHrP production in a time-dependent manner, also by Day 1. A 125I-TGF-beta 1 binding study revealed that HOC-21 cells expressed a high affinity for the TGF-beta 1 receptor. CONCLUSION The suppression of human ovarian cancer cell proliferation by TGF-beta 1 might be involved in the production of PTHrP.

Activin facilitates the follicle-stimulating hormone (FSH) secretion and FSH-beta mRNA level in rat pituitary cell culture (1–4). These changes in FSH secretion are rapid and suggest that activin may have important roles in the dynamic changes of FSH secretion throughout the reproductive cycle. Moreover, activin has been shown to inhibit pituitary growth hormone (GH), prolactin (PRL) (5), and adrenocorticotropic hormone (ACTH) (6) release. In addition to these actions on hormone secretion from the pituitary, activin is known to induce differentiation of erythroid cells (7) and cause mesoderm induction in Xenopus (8). Activin is considered to be a member of the transforming growth factor-beta (TGF-beta) superfamily by these multiple effects.

mTOR is an important factor for cell proliferation. It is reported that overexpression of mTOR is associated with poor prognosis in many cancers. We examined mTOR expression in cervical cancer and association with clinico-pathological findings. 34 patients were examined about mTOR expression by immuno-histochemistry. The result was classified into 3 categories, i.e negative, weak positive and strong positive. 25 patients were treated with concurrent chemoradiotherapy, 8 ptients were treated with hysterectomy and one patient was treated with chemotherapy as main treatment. Overexpression of mTOR was recognized in 24(71%) patients. The frequency of parametrial invasion and lymphnode metastasis in mTOR positive and negative were 17/24 (71%) versus 3/10 (30%) (p=0.027) and 14/24 (58%) versus 2/10 (20%) (p=0.041). In 25 cases treated with radiotherapy, mTOR overexpression was seen in 19 cases (76%). Averages of tumor diameter in mTOR positive and negative cases were 4.8 cm and 5.0cm respectively (n.s). In 5 local recurrent cases treated with chemo-radiotherpy, tumor diameters of pre-treatment were 0, 3.5, 5.5, 7 and 7cm. Small tumor was included in local recurrence in mTOR positive cases. It is suggested that mTOR overexpression promotes lymphnode and parametrial metastasis and is associated with radio-resistance. In case of mTOR overexpression, we need to consider treatment by hysterectomy if the tumor is resectable or usage of mTOR inhibitor at chemo-radiation, and control of mTOR expression is important in treatment of cervical cancer. Citation Format: Masato Nishimura, Kanako Yoshida, Hiroyuki Furumoto, Minoru Irahara. mTOR overexpression is associated with radio-resistance and worse prognosis of cervical cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 433. doi:10.1158/1538-7445.AM2013-433

Arachidonate 12-lipoxygenase, which oxygenates positions 12 and 13 of arachidonic and linoleic acids, is present in porcine anterior pituitary cells. Colocalization of the 12-lipoxygenase with various pituitary hormones was examined by immunohistochemical double-staining using antibodies against 12-lipoxygenase and various anterior pituitary hormones. Under light microscopy, approximately 7% of the cells producing luteinizing hormone (LH) and follicle-stimulating hormone (FSH) were positive for 12-lipoxygenase, whereas the enzyme was detected in less than 2% of the cells producing thyrotrophin, prolactin, growth hormone (GH), and adrenocorticotrophin. In an attempt to examine the participation of 12-lipoxygenase metabolites in pituitary hormone release, we incubated the primary culture of porcine anterior pituitary cells with 12-hydroperoxy-arachidonic acid or 13-hydroperoxy-linoleic acid. Significant stimulation of LH and FSH release by these hydroperoxides was observed at 10 μm in a time-dependent manner. At doses around 10 μm these compounds produced responses of similar magnitude to 1 nm gonadotrophin-releasing hormone (GnRH), but higher concentrations (30 μm) of the compounds were required for GH release. In contrast, 12-hydroxy-arachidonic and 13-hydroxy-linoleic acids were almost ineffective. Furthermore, the gonadotrophin release by 1 nm GnRH was inhibited by nordihydroguaiaretic acid (a lipoxygenase inhibitor) with an IC50 of about 5 μm. Thus, the hydroperoxy (but not hydroxy) products of 12-lipoxygenase may be involved in the release of pituitary hormones especially LH and FSH. Journal of Endocrinology (1996) 148, 33–41

Yoneda N, Irahara M, Saito S, Uemura H, Aono T. Usefulness of recombinant human prolactin for treatment of poor puerperal lactation in a rat model. Eur J Endocrinol 1995;133:613–7. ISSN 0804–4643 Recombinant human prolactin (r-hPRL) was produced by a line of murine C127 cells transfected with human PRL gene. To assess the biological efficacy of r-hPRL in vivo, we studied its influence on milk secretion using a rat model in which lactation was reduced by bromocriptine treatment. Puerperal rats were injected daily for 9 days after delivery with bromocriptine or bromocriptine plus r-hPRL, and lactational performance was assessed by weighing the pups. The concentrations of rat and human PRL in rat serum were measured by specific radioimmunoassays and the mammary glands were examined on postpartum day 10. Daily injection of bromocriptine (0.1 mg/rat) significantly reduced the endogenous level of rat PRL and impaired the weight gain of the pups. Administration of r-hPRL increased the serum level of human PRL. Daily injections of r-hPRL (50 μg/rat, twice a day) restored lactational performance and significantly increased the weight of the pups. The detrimental effect of bromocriptine on the mammary glands, assessed by both weight and histological appearance, was reversed by administration of r-hPRL. These results demonstrate that r-hPRL is biologically active in vivo and replacement therapy of r-hPRL is effective in improving the lactational performance in bromocriptine-treated rats, and also that r-hPRL may be useful for the treatment of women with poor lactation. Naoto Yoneda, Department of Obstetrics and Gynecology, School of Medicine, The University of Tokushima, 3-18-15 Kuramoto, Tokushima 770, Japan

Follistatin, an activin-binding protein, is able to neutralize the various activities of activin by forming an inactive complex with it. The widespread tissue localization of follistatin is very similar to that of activin, which suggests that it plays a local modulatory role in the various paracrine/autocrine actions of activin. We detected significant activin-binding activities in human follicular fluid and placental homogenates, although they were much lower than those in porcine and bovine follicular fluids, which raised the possibility that follistatin is present in human follicular fluid and placenta. Therefore, we attempted to identify the protein molecules responsible for this activin-binding activity in human follicular fluid and placental homogenates. Human follicular fluid, collected from in vitro fertilization patients, was processed by affinity chromatography successive steps on sulfated gel matrices and reverse phase high performance liquid chromatography (HPLC). The final HPLC yielded abundant follistatin and almost equimolar amounts of activin-A, -AB, and -B. The follistatin protein showed characteristic multiple bands when analyzed using sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The ability of each band to bind activin specifically was determined by activin binding assay and ligand blotting analysis. Several pieces of evidence, including the immunoblotting analysis and functional assay results, demonstrated the presence of three activin isoforms, A, AB, and B, in the follicular fluid. In contrast, human placental homogenates were found to contain follistatin and activin-A proteins only. Activin-AB and -B were not detected in any HPLC fraction, indicating that activin-A is the major form of activin in the human placenta. The present data indicate that the three activin isoforms and multiple forms of follistatin exist in human follicular fluid, and the activin-A isoform and follistatin exist in human placenta. They suggest that the physiological functions of the activin isoforms during embryonic development differ and that follistatin plays a functional role in the local control system(s) that regulates human reproduction.

The relationship between maternal plasma levels of beta-endorphin (beta-Ep) during labor and various obstetrical factors was investigated in 115 healthy pregnant women. beta-Ep was determined by radioimmunoassay using double-antibody RIA kit (INCSTAR Corporation, Stillwater, M'S.). The results were as follows: (1) The primiparous women showed a significant increase of maternal plasma levels of beta-Ep at delivery compared with the multiparous women. In addition, the group of women whose Bishop's score at the onset of labor was 5 points or less showed a significant increase of maternal plasma levels of beta-Ep at delivery compared with that in the group of women whose Bishop's score was 6 points or more. (2) The increase in maternal plasma levels of beta-Ep during the first and the second stage of labor was significantly higher in obese women (pre-pregnancy BMI > or = 24) than in normal weight women (pre-pregnancy BMI < 24). In normal weight women in pre-pregnancy, the group of women whose weight gain during pregnancy was 11kg or more showed a significantly higher increase of beta-Ep compared with that in the group of women whose weight gain was less than 11 kg. These results suggest that a stressful delivery caused a significant increase of maternal plasma levels of beta-Ep during labor. Moreover, obesity and marked weight gain during pregnancy caused a remarkable increase in beta-Ep probably due to latent dystocia.

The responses of prolactin (PRL) and oxytocin (OT) to suckling and breast massage were examined in lactating women. The suckling group showed increase in frequency of pulsatile release of OT and increase of the plasma PRL level. In contrast, the breast massage group showed a significant, but not a pulsatile increase in the plasma OT level and no increase in the plasma PRL level. These findings suggest that suckling causes both milk production and milk ejection, while breast massage causes only ejection of milk already stored in acini, and that prolactin release is not related to increase of the oxytocin level itself, but to its pulsatile release.

The effects of 2-buten-4-olide (2-B40), an endogenous feeding suppressant, on the secretion of luteinizing hormone (LH) were studied in ovariectomized rats. Intraperitoneal (ip) administration of 2-B40: adult female ovariectomized Wistar rats were given daily ip injections of solution containing 2-B40 at 0, 50 or 100 mg/kg body wt for 14 days. This ip treatment with 2-B40 significantly decreased the mean LH concentration and pulse frequency of LH. Intravenous (iv) administration of 2-B40: a solution of 2-B40 (50 or 100 mg/kg body wt) was slowly injected through an intra-atrium catheter and blood samples were taken every 6 min for 2 h. This iv treatment significantly suppressed the LH pulse frequency but had no significant effect on the LH amplitude or mean LH. Injection of 2-B40 into the third cerebroventricle: the injection of 2-B40 into the third cerebroventricle of freely moving rats decreased the mean LH concentration and the frequency and amplitude of LH pulses. Third cerebroventricle injection of a corticotropin-releasing factor (CRF) receptor antagonist before third cerebroventricle injection of 2-B40: the specific CRF receptor antagonist α-helical-CRF (9–41) was injected into the third cerebroventricle of ovariectomized rats before injection of 2-B40. Administration of 2-B40 into the third cerebroventricle significantly decreased the mean LH, concentration and pulse frequency. Third cerebroventricle injection of the CRF antagonist at 50 μg/rat and then 2-B40 also resulted in significant suppression of the mean LH concentration and pulse frequency. These findings suggest that 2-B40 may suppress gonadotropin-releasing hormone secretion in ovariectomized rats and that its effect in decreasing LH pulses may not be caused through the CRF pathway.

PROBLEM: Autoantibodies to zona pellucida (ZP) have been implicated as a cause of infertility in woman by three lines of clinical and laboratory evidence. METHOD: First, positive anti-zona activities, as assessed by the passive hemagglutination reaction (PHAR) using bovine red blood cells sensitized with porcine zona antigen, were detected in 45 of 1,872 serum samples collected from infertile women, but in only three (two nonpregnant and one pregnant) of 592 serum samples from control subjects. Their incidence in 320 women with unexplained infertility (5.6%) was much higher than that in 1,552 women with infertility of known cause (1.7%, P < 0.01), which was comparable to the incidence observed in 193 age-matched fertile nonpregnant and pregnant women (1.5%, P < 0.05). None of the serum samples from 292 age-matched adult men and 100 children of between 5 and 10 years old gave a positive PHAR. Second, follow-up study for a minimum of 2 years with treatment revealed that no pregnancy occurred in 11 infertile women with a consistently positive PHAR, and only three pregnancies in 19 infertile women with a fluctuating positive PHAR. Third, three of seven serum samples from women who gave a consistently positive PHAR produced strong immunofluorescence reactions on human ZP even after adsorption with porcine and human AB blood cells. Pre-exposure of human ZP to the sera showing positive immunofluorescence, including that of one patient undergoing an in vitro fertilization program, greatly diminished the number of spermatozoa of normal quality that bound to and penetrated across human ZP. CONCLUSIONS: The observed high incidence of anti-zona activities and long-term resistancy to treatment in women with unexplained infertility may be closely correlated with inhibition of sperm-egg interaction by anti-zona autoantibodies produced in these women.

Introduction and aims The use of complementary and alternative medicine (CAM) has been increasing in Japan as well as in other countries. Among various modalities, traditional Japanese herbal medicine (Kampo) is rather well recognized and is integrated into the Japanese health-care system. On the other hand, many Japanese physicians assign a limited role in CAM except Kampo, and only a few public university hospitals, such as Osaka University and Kanazawa University, have a CAM clinic in Japan. At Tokushima University Hospital, an academic hospital in the Shikoku district, we started CAM consultation clinic in July 2007 to give consumers scientific information of CAM by specialists. Here we report the present activity of the clinic. Methods and results The main issues that we deal are health foods including dietary supplements and Kampo. At present, we do not accept consultations for other types of CAM, such as mind–body interventions or energy therapies. Two nutritionists and four pharmacists (professors and associate professors of the University of Tokushima Graduate School) and two pharmacists belonging to the Tokushima University Hospital play roles in the CAM consultation clinic. From July 2007 to June 2008, we dealt 43 cases. The underlying status for consultation were malignant diseases (23%), cardiovascular diseases (21%), collagen vascular diseases (9%), allergic diseases (5%), other chronic diseases (26%) and no specific underlying diseases (general well-being) (16%). The area for consultation was health foods including dietary supplements (37%), Kampo (30%) and others (33%). Conclusion For an effective and safe use of CAM, close contact between consumers, consultation team and physicians and that between primary care institutions and University Hospitals are necessary, and we are going to widen our activity to catch up with these issues.

The effects of 2-buten-4-olide, an endogenous feeding suppressant, on the estrous cycle and LH secretion were studied to determine the influence of this compound on reproductive function. Estrous cycling female Wistar rats were treated ip with 2-buten-4-olide (0, 30 or 100 mg·kg−1·day−1) for 2 weeks. Treatment with 100 mg·kg−1·day−1delayed the estrous cycle. 2-Buten-4-olide increased the pituitary content of LH (1651.3±164.4 vs 927.7±65.1 ng/pituitary; p−4 mol/l) suppressed the pituitary response to GnRH (2 μg/l) (percent increase at 50 min after start of GnRH stimulation: 180±47 vs 406±66%; p

We present a case of Ebstein’s anomaly with tri-phasic inferior vena cava (IVC) and ductus venosus (DV) forward flow pattern. Pulsed tissue Doppler imaging (TDI) were performed to elucidate the relationship of the atypical venous flow and the ventricular wall motions. A 32 year-old nulliparous woman conceived a monochorionic-diamniotic twin pregnancy by IVF. On a routine fetal sonography at 14 weeks of gestation, Twin A showed cardiomegaly (cardio-thoracic ratio: 53%) and severe tricuspid regurgitation (TR). Another twin was sonographically unremarkable. With advance of gestation, fetal echocardiography on Twin A revealed right atrial enlargement, severe holosystolic TR (1.3m/sec), the enlarged sail-like anterior leaflet of tricuspid valve (TV), and retrograde pulmonary artery and ductal flow in the absence of pulmonary regurgitation. Posterior and septal leaflets of TV were not detected. For sonographic findings stated above, Ebstein’s anomaly and pulmonary atresia was prenatally diagnosed. Pulsed Doppler examination showed tri-phasic IVC and DV forward flow pattern. Pulsed TDI recording of the right atrioventricular valve ring showed abnormal shortening after S wave from late systolic phase to isovolumic relaxation phase of the cardiac cycle. This abnormal shortening synchronized with the second wave of IVC and DV forward flow, and the left ventricle showed the same shortening. There were no sign of fetal arrhythmia or hydrops fetalis. A Cesarean section was performed at 36 weeks of gestations. Twin A was a female infant weighing 2290g with Apgar scores of 2 and 6 at 1 and 5 minutes, respectively. Postnatal diagnosis was the same as prenatal assessment. Starnes operation was performed at 6th day after birth. But she died at the 7th day because of cardiac failure. In conclusion, we detected tri-phasic IVC and DV forward flow pattern in the fetus with Ebstein’s anomaly. We speculate that the abnormal ventricular motion caused the atypical venous flow pattern.

We investigated the effects of Toki-shakuyaku-san and its crude ingredients in relation to the secretion of a cytokine-induced neutrophil chemoattractant, CINC/gro, interleukin-1 beta (IL-1 beta) and tumor necrosis factor-alpha (TNF alpha) in the ovulatory process. Toki-shakuyaku-san significantly (p < 0.01) stimulated the secretion of 17 beta-estradiol but did not stimulate the secretion of progesterone in cultured whole ovarian dispersates. Toki-shakuyaku-san enhanced the secretion of CINC/gro in a dose-dependent manner and the production of CINC/gro at concentrations of 10 and 100 micrograms/ml of Toki-shakuyaku-san increased significantly (p < 0.01). Toki-shakuyaku-san also enhanced secretions of both IL-1 beta and TNF alpha, which are known to stimulate the secretion of CINC/gro in the ovulatory process. The production of TNF alpha increased significantly (p < 0.05) with 10 and 100 micrograms/ml of Toki-shakuyaku-san. Atractylodis Lanceae Rhizoma, Cnidii Rhizoma, Angelicae Radix, Paeoniae Radix and Alismatis Rhizoma, which are crude ingredients of Toki-shakuyaku-san, significantly (p < 0.01) enhanced the secretion of CINC/gro at concentrations of 100 micrograms/ml. The results of this study show that Toki-shakuyaku-san can stimulate the secretion of 17 beta-estradiol and stimulate the ovulatory process by stimulating the production of CINC/gro, IL-1 beta and TNF alpha in vitro. As a treatment for ovulatory disorders, Toki-shakuyaku-san may have stimulatory effects on both steroidogenesis and the ovulatory process.

To assess the effect of PRL on fertilization and cleavage of oocytes, PRL levels in serum and follicular fluid of normoprolactinemic women who had undergone in vitro fertilization and embryo transfer (IVF-ET) were determined, and the relationship between PRL levels and fertilization and cleavage rates was studied. A transient increase in serum PRL at the ovulatory phase was found in about half of the IVF-ET patients during clomiphene stimulation, and increased levels of PRL in serum and follicular fluid at the ovulatory phase may suppress the fertilization and cleavage of oocytes.

Two cases of peritonitis carcinomatosa with gynecological cancer, which did not respond to conventional treatment, were treated by intraperitoneal adoptive immunotherapy with allogeneic or autologous lymphokine-activated killer (LAK) cells plus interleukin-2. In case 1, reduction of acute ascites and disappearance of malignant cells from the peritoneal fluid and decreased levels of tumor markers (CA12-5 and CA19-9) were demonstrated during the treatment. In case 2, which also received additional treatment for pleuritis carcinomatosa, reduction of ascites and pleural effusion and disappearance of malignant cells from the pleural fluid were noted. In these cases allogeneic LAK therapy was well tolerated. This therapy did not prolong the patients' lives, but the findings indicate that it had appreciable local antitumor effects on peritonitis and pleuritis carcinomatosa in these patients with gynecological cancer.

A simple method of detecting anti-zona activities in human sera has been developed by passive hemagglutination reaction ( PHAR ) using bovine erythrocytes as indicator cells coated with purified porcine zona substance. By this test, 8 of 88 serum samples (9.1%) from infertile women gave positive reactions, whereas only 1 of 90 control sera (1.1%). A positive immunofluorescence (IF) assay confirmed the results of the PHAR in seven of the eight positive samples. Positive PHAR and IF on porcine zonae were abolished after adsorption with porcine erythrocytes, but IF on human zonae was retained by four of eight serum samples, indicating the presence of anti-zona autoantibodies in these infertile sera that were directed to an antigen(s) proper to human zonae.

Serum samples with sperm-immobilizing antibody activity from six women were examined for ability to block sperm-egg interaction by a zona penetration test where human follicular ova matured in vitro were used. Exposure of spermatozoa from a fertile healthy donor to the sera impaired binding to and penetration through the zona pellucida of the spermatozoa completely in five cases and incompletely in one case. Successful fertilization in vitro was achieved by using fetal cord serum instead of autoserum of the patient included in the in vitro fertilization and embryo transfer program. These results suggest that interference with sperm-egg interaction may be an additional mechanism of infertility that is caused by antisperm antibodies.

Measurement of somatostatin (SRIF) by RIA in porcine ovarian tissues after removal of the follicular fluid revealed the existence of immunoreactive (IR) SRIF at concentrations of 2.2 ± 0.6 ng/g wet weight (mean ± sd). On gel chromatographic analysis of ovarian extracts one major and two minor components with SRIF-like immunoreactivities have been detected. The major component comprised 59.5 ± 15.1% of the total IR-SRIF and was coeluted with synthetic tetradecapeptide (SRIF 14). The two other components seemed to have almost the same molecular weights as synthetic octacosapeptide (SRIF 28) (25.1 ± 10%) and pro-somatostatin (15.4 ± 5.5%), respectively. It is concluded that SRIF in porcine ovaries is heterogeneous, comprising SRIF 14 as a major component, as well as larger forms of SRIF which probably represent SRIF precursors.

A comparative study on anti-zona antibody activities in the sera from clinically defined categories of patients registered at the WHO Reference Bank for Reproductive Immunology was performed at five different laboratories with different detection methods. Considerably higher incidences of positive reactions were detected by immunofluorescence on porcine zonae in infertile women (16.3%) than in control subjects (7.1%). A similar proportion of positives was found by radioimmuno-binding assay (RIBA) using porcine zona antigen preparation in the infertile group (13.0%) but not in the female control group (0%), giving an indication of the specificity of this test. It is noteworthy that high incidences of positives were observed by RIBA with sera from male subjects with unexplained sterility, vasectomy and aspermatogenesis. A test system of passive hemagglutination reaction (PHAR) using purified porcine zona substance as antigen gave a low but slightly higher incidence of positives in infertile (3.1%) than in control sera (0.9%). No positive reactions were observed with infertile and control sera by another PHAR or by radioimmunoassay using an antigen preparation common to the two test systems. Anti-zona activities in these sera were therefore seen to vary, depending largely upon the detection systems and the antigen preparations.

The losses of blood group antigens A, B, and H in carcinoma tissue of the uterine cervix were studied by the avidin-biotin-peroxidase complex (ABC) method and the relations of these losses to invasion and dedifferentiation of primary cancer were examined. The incidence of cases showing complete loss of A or B antigen increased in proportion to the progression of cancer, but in most cases even of invasive cancer, H antigen, the precursor of A and B antigens, was detected. Complete loss of H antigen was not demonstrated in well-differentiated keratinizing invasive carcinomas, but was seen in 15% (15/101) of the cases of large cell non-keratinizing type cancer and 50% (8/16) of those of small cell non-keratinizing type cancer. No relationship was found between losses of A, B, and H antigens and parametrial spread of carcinoma or metastasis to the pelvic lymph nodes, but the incidence of death within 2 years after hysterectomy was higher in H antigen-negative cases than in H antigen-positive cases. These results indicate that loss of A and B antigens depends on some activity of invasion of cancer, while loss of H antigen strongly indicates dedifferentiation of cancer cells and also may indicate a poor prognosis.

To investigate the physiological importance of somatotrophin release-inhibiting factor (SRIF), effects in vitro of synthetic SRIF 14 on germinal vesicle breakdown (GVB) of cultured porcine follicular ova were studied. The proportion of ova with GVB decreased gradually and significantly with increasing concentrations of SRIF 14 in the range from 6 × 10−12 to 6 × 10−7 m during a 22 h period of culture. The inhibitory effect was apparent for the period between 14 and 22 h in the course of culture but was reversed by a concomitant addition of anti-SRIF to the medium. Neither synthetic oxytocin, vasoactive intestinal polypeptide nor substance P exerted any inhibitory or stimulatory action on GVB. These results suggest a limited but definite inhibitory action of SRIF on GVB of porcine ova.

The role of monosaccharides in human fertilization was studied by testing their effects on penetration of spermatozoa into mature human oocytes (zona penetration test). When oocytes were pretreated with concanavalin A, wheat germ agglutinin, or Ricinus communis agglutinin-I at a concentration of 100 micrograms/ml, no spermatozoa were found to bind to or penetrate through the zona pellucida. Penetration of spermatozoa was restored when the zona pellucida pretreated with wheat germ agglutinin and Ricinus communis agglutinin-I were rinsed with N-acetyl-D-glucosamine (wheat germ agglutinin inhibitor) and D-galactose (Ricinus communis agglutinin inhibitor), respectively. Conversely, the blocking effect of concanavalin A on sperm penetration was not reversed by treatment with D-mannose (concanavalin A inhibitor). Furthermore, pretreatment of spermatozoa with D-mannose (50 mmol/L) completely inhibited sperm penetration through the zona pellucida. However, sperm penetration was clearly demonstrated when the zona pellucida was pretreated with D-mannose. These data suggest that D-mannose residues are essential in, or sterically closely related to, the sperm receptor site on the human zona pellucida.

Background Clinical trials play a central role in the establishment of clinical evidence, and the important role of clinical research coordinators (CRCs) in various processes of clinical trials is now widely recognized. In Japan, many CRCs work under the discretion of their hospital and support clinical trials in various areas. Modification of CRC activity pursuant to the types of clinical trials may make roles of the CRC more effective and meaningful. In the present study, we examine the dedicated role of the CRC considering the specialty of a registration trial of a drug for surgical patients used during the operation period. Methods In 2006, we had a chance to support a registration trial of a drug for surgical patients used during the operation period. Regarding the mental and emotional status of possible participants in the present registration trial, we collected data from the perspective of CRCs by focus group interviews involving four CRCs working under the discretion of Tokushima University Hospital. The four CRCs were all nurses and had 7, 4.5, 1, or 0.5 years experience as CRCs, respectively. Results In contrast to clinical trials of drugs for chronic diseases, these often anxious patients must decide whether or not to enter the trial simultaneously with the decision to undergo surgery itself, and all in a relatively limited time after receiving explanation of the trial. Therefore, special attention should be paid to the mental and emotional status of possible participants. Additionally, the cooperation of the relatively large surgical and nursing staff becomes important. In such situations, the following contributions of CRCs were considered to be useful for the harmonious procedure of clinical trials: 1) providing a precise explanation of the trial to the participant and key persons, 2) understanding the needs of the investigators and appropriately assigning themselves roles, and 3) communicating between the investigators and surgical and nursing staff. Conclusion Further study is warranted to evaluate the benefit of the intervention provided by dedicated CRCs in running high quality clinical trials involving surgical patients.

Background Clinical trials in the general practice setting are important for providing evidence on the effectiveness and safety of different agents under various conditions. In conducting these trials, the participation of physicians and patient recruitment are important issues. Various investigations in the literature have reported views and attitudes of physicians on various types of clinical trials. Nevertheless, there is still little information concerning physicians participating in a clinical trial and among them, those who could not recruit any patients (unsuccessful physician recruiters). Methods In 2003, we collaborated in a large-scale multicenter study of Japanese hypertensive patients (COPE Trial). In Tokushima University Hospital and 18 other medical institutions, we investigated the views and attitudes of unsuccessful physician recruiters in comparison with successful physician recruiters, using a questionnaire. Results The questionnaire was provided by mail to 47 physicians and 27 (57%) responded. The response rate was 79% for successful physician recruiters compared to 43% (P = 0.014) for unsuccessful physician recruiters. More successful physician recruiters (73%) than unsuccessful physician recruiters (42%) stated they had participated and enrolled patients in previous multicenter clinical trials. A significantly higher number of successful physician recruiters than unsuccessful physician recruiters (42%; P = 0.040) considered the presence of a support system with clinical research coordinators (CRC) as the reason for participation (80%). A large number of unsuccessful physician recruiters experienced difficulty in obtaining informed consent (67%), whereas a significantly smaller number of successful physician recruiters experienced such difficulty (20%; P = 0.014). The difficulties experienced by unsuccessful physician recruiters in the trial were as follows: inability to find possible participants (100%), difficulty in obtaining informed consent (58%), cumbersome procedures (58%), difficulty in long-term follow up (33%), and insufficient tools for explanation and obtaining informed consent (8%). Conclusion This survey showed that successful physician recruiters consider a support system with CRC of value, and that they are skillful in obtaining informed consent. These views and attitudes may have originated from past experience involving clinical trials. In this regard, we need to develop an infrastructure to enlighten physicians on this support system for the promotion of clinical trials.