Hiroe Ito

Bone morphogenetic protein 2 may be a good predictor of success in oocyte fertilization during assisted reproductive technology

The aim of this study was to determine whether the bone-morphogenetic proreins-2 (BMP-2) or-7 (BMP-7) levels in follicular fluid (FF) are associated with success in oocyte fertilization during assisted reproductive technology (ART) treatment. Twenty-four cycles in 24 patients who underwent oocyte retrieval in Sugiyama clinic were included in this retrospective study. The patients were divided into two groups according to the success or failure of fertilization. FF samples were obtained from a single follicle in each patient, and the levels of BMP-2, BMP-7, Anti-Müllerian hormone (AMH), estradiol and progesterone from FF were measured, and evaluated in relation to the ART outcomes. The BMP-2 levels correlated positively with the AMH levels in FF (r2 = 0.4928), but there was no statistically significant difference between BMP-7 and AMH levels. The BMP-2 and BMP-7 levels had no relation with either progesterone or estradiol levels, but BMP-2 levels in the fertilized group were significantly higher than those in the unfertilized group (P < 0.05). The BMP-2 levels in FF positively correlated with the AMH levels in FF, and those in the fertilized group were significantly higher than in the unfertilized group. Therefore, the BMP-2 levels in FF could be a predictive marker for fertilization.

Expression of leukemia inhibitory factor in the endometrium in abnormal uterine cavities during the implantation window

OBJECTIVE To investigate the expression of the implantation factors leukemia inhibitory factor (LIF), interleukin-11 (IL-11), and IL-6ST/gp130 in the endometrium to examine the relationship between loss of implantation and abnormal uterine cavity. DESIGN Case-control study. SETTING Department of Obstetrics and Gynecology, Tokyo Medical University. PATIENT(S) Subjects comprised 41 patients in the abnormal uterine cavity group who underwent resection of a uterine submucosal myoma or an endometrial polyp by transcervical resectoscopy (TCR) and 18 patients in the control group who underwent laparoscopic surgery. INTERVENTION(S) In the abnormal uterine cavity group, endometrial tissue specimens were obtained before resection under hysteroscopy. In the control group, endometrial tissue specimens were obtained by curettage at the time of laparoscopic surgery. MAIN OUTCOME MEASUREMENT(S) We divided the patients into four groups according to menstrual cycle and measured the endometrial expression of LIF, IL-11, and IL-6ST/gp130 with the use of quantitative real-time reverse-transcription polymerase chain reaction. RESULT(S) Significant increases in LIF and IL-11 mRNA expression were recognized during the midsecretory phase of the menstrual cycle in the control group. However, no difference in IL-6ST/gp130 mRNA expression was observed in any phase during the menstrual cycle in either group. In terms of LIF and IL-11 mRNA expression at the midsecretory phase, the abnormal uterine cavity group showed a significantly decreased LIF mRNA expression compared with the control group. CONCLUSION(S) LIF mRNA expression was significantly decreased in abnormal uterine cavities during the midsecretory phase, indicating that endometrial cavity defects are a possible cause of poor reproductive outcomes.

The ubiquitin-conjugating enzyme E2-EPF is overexpressed in cervical cancer and associates with tumor growth

We found that the ubiquitin-conjugating enzyme E2-EPF mRNA is highly expressed in cervical squamous cancer relative to normal tissues and its expression levels positively correlate with clinical stage. Reduction of E2-EPF protein levels by >80% using shRNA decreases the expression levels of HIF-1α, and the proliferation, invasion and tumorigenicity of SiHa, a cervical squamous cancer cell line. E2-EPF knockdown also increases the chemosensitivity to topoisomerase I inhibitor (topotecan) and II (etoposide and doxorubicin). Our results suggest that E2-EPF is associated with the growth and aggressivity of cervical tumor cells. Targeting the E2-EPF pathway may have potential clinical applications for the treatment of cervical cancer.

Establishment of three cisplatin-resistant endometrial cancer cell lines using two methods of cisplatin exposure

AbstractUsing the endometrial cancer cell line EI established in our department, we attempted to establish cisplatin (CDDP)-resistant cell lines by incremental exposure and high concentration exposure methods. Three CDDP-resistant cell lines were isolated, which could be distinguished by morphological differences. 1.Upon acquiring CDDP resistance, the cells tended to become small and grow in a floating state. This tendency was especially marked when using incremental exposure method. Using the incremental exposure method, a cell line obtained by isolating and culturing only adherent cells was designated EICR-Ia, and a cell line established by culturing only floating cells was designated EICR-If. A cell line obtained by the high concentration exposure method was designated EICR-II.2.Upon acquiring CDDP resistance, tumor markers such as TPA and LDH increased, while proliferative capability of the cells was lowered.3.The invasion capability was diminished in EICR-If cells, but was increased in EICR-Ia and EICR-II cells.4.Following exposure to CDDP, the intracellular platinum concentrations were markedly elevated in EI and EICR-If cells, whereas the increase was mild in EICR-Ia and EICR-II cells and the concentration was lower than that in parent EI cells.5.Studies of drug resistance gene expression revealed increased expression of MDR1, GSTπ, and Topo-II in EICR-If cells; increased expression of GSTπ in EICR-II cells; but no expression of any of the genes in EICR-Ia cells.6.Analyses of cancer- and apoptosis-related genes showed increased expressions of Bcl-2, c-Myc, p53, and ICE in EICR-If cells.7.Upon acquiring CDDP resistance, sensitivity to mitomycin and adriamycin decreased, but sensitivity to etoposide and 5-fluorouracil increased. The findings indicate that the mechanisms of CDDP resistance are different in the three cell lines.

Omega-3 fatty acid supplementation for expectant mothers with depressive symptoms in Japan and Taiwan: An open-label trial.

SLEEP PARALYSIS (SP) IS a state of involuntary immobility occurring at sleep onset or offset. Most studies show that 18–40% of the general population has experienced SP at least once in their lifetime. No study has directly assessed rates of SP across groups in one country, which is the goal of the current studies. Study I explored rates of lifetime occurrence of SP in a sample of self-identified ethnic individuals in Denmark (n = 50; male = 38%; age: M = 35.4, SD = 12.5, range = 18–56; non-Caucasian members of ethnic groups, including Asians, Middle Easterners, and North Africans) compared to rates among all self-identified non-ethnic Danish Caucasian individuals (n = 173; male = 49.7%; age: M = 29.1, SD = 10.8, range = 19–70). Study II compared the lifetime frequency rates of SP of ethnic Danish individuals (n = 23; male = 62%; age: M = 27.3, SD = 11.8, range = 19–55) with that of non-ethnic Danish individuals (n = 35; male = 53%; age: M = 23.7, SD = 6.9, range = 18–51; all Caucasian). All participants were fluent in the Danish language. The institutional review board of the first author’s former institution, the American University in Cairo, approved these studies. In Study I, participants were screened orally for once-in-alifetime occurrence of SP, using the Sleep Paralysis Questionnaire (SPQ). Convenience sampling was used to recruit participants from different locations and community settings. We found that a greater number of ethnic Danes had experienced SP at least once in their lifetime compared to non-ethnic Danes, namely, 44% (22/50) versus 20% (35/173), χ = 12.2, P < 0.001. In Study II, lifetime frequency of SP was assessed orally using the SPQ; all participants had experienced SP at least once. They were recruited through referrals by colleagues and acquaintances as well as announcements in community settings. The convenient sampling method, ‘chain referral,’ was utilized to increase sample sizes. We found that ethnic Danes had higher lifetime frequency rates of SP relative to nonethnic Danes, M = 8.9 (SD = 11.3) versus M = 4.1 (SD = 5.7), t(29) = 1.9, P < 0.05. What is striking about the current findings is the degree to which SP rates differ in the two groups: the non-Caucasian ethnic Danish individuals were more than twice as likely to experience SP as compared to non-ethnic Danes. Moreover, members of Danish ethnic groups had more than twice as many SP episodes in a lifetime; that is, a great difference in SP rates for groups who belong to the same cultural framework, but differ with respect to ethnic heritage (nonCaucasian vs Caucasian). These results stress the importance of clear ethnic breakdown when examining rates of SP. DISCLOSURE STATEMENT

Efficient production of trophoblast lineage cells from human induced pluripotent stem cells

Human induced pluripotent stem cells (hiPSCs) are potentially useful in both clinical applications and basic biological research. hiPSCs can differentiate into extra-embryonic cells in the presence of BMP4. However, the differentiation potential of hiPSCs can be affected by culture conditions or genetic variation. In this study, we investigated the effect of various BMP4 concentrations on the expression states of trophoblast markers and the optimal conditions for trophoblast induction. A high-fidelity gene expression assay using hiPSC lines showed that the expression levels of various trophoblast marker genes, such as KRT7, GCM1, CGB, and HLA-G, were upregulated by BMP4 in a dose-dependent manner in all types of hiPSCs used in this study. Treatment with high doses of BMP4 for prolonged periods increased the ratio of cells with trophoblast markers irrespective of the presence of bFGF. We found that the expression states of major pluripotency- and differentiation-related protein-coding genes in BMP4-treated cells depended on culture conditions rather than donor cell types. However, miRNA expression states were affected by donor cell types rather than BMP4 dose. Furthermore, the effect of the presence of bFGF on differentiation potential of KRT7-positive cells differed among iPSC types. Mechanistically, chromatin states around KRT7 promoter regions were comparable among the iPSC types used in this study, indicating that hiPSC chromatin state at these regions is not a parameter for cytotrophoblast differentiation potential. In conclusion, the optimal conditions for trophoblast differentiation from hiPSCs differ according to parental cell line.

Telomerase activity in human trophoblast

Summary Recent reports have suggested that many proliferating normal cells express telomerase activity, although telomerase activity was previously considered to be specific for cancer cells. This fact draws attention to the investigation of telomerase activity in trophoblast cells which have characteristics of proliferation, differentiation and invasion. Sub-populations of trophoblast cells were separated by a modification of Klimans method and magnetic cell sorting with antibodies of integrins. Telomerase activity was assayed semi-quantitatively using fluorescence-based TRAP. Cytological detection of telomerase activity was also examined by the newly developed in situ TRAP procedure and the expression of human TERT was detected by RT-PCR. Telomerase activity was detected in 42% of normal placental tissues, and demonstrated decreased activity with gestational age. Telomerase activities was closely correlated with expression of TERT. In sub-populations of CT separated from placenta from early and term gestation, telomerase activity was predominantly detected in columnar CT from early placenta. Fluorescence signals after in situ TRAP were observed in the trophoblastic cell layers in early placenta as well as isolated CT, but not in term placenta. These results suggest that telomerase activity correlates with the regulation of trophoblastic cell proliferation and in situ TRAP is useful for the detection of telomerase positive cells.

Ultrasound elastography can detect placental tissue abnormalities

Abstract Background In this prospective cohort study, we examined the utility of elastography to evaluate the fetus and placenta. Patients and methods Pregnant women in their third trimester of pregnancy, by which time the placenta has formed, were included in this study. A total of 111 women underwent ultrasound examinations, including elastography. Elastographic evaluation was performed using two protocols. First, the placental index (PI) was measured, which quantitatively assesses the hardness of tissue. Second, regions of interest (ROI) were categorized into 3-step scores according to the frequency of the blue area (hardness of placental tissue score [HT score]), which is a qualitative method. After delivery, 40 of the 111 placentas were pathologically examined. Results The average PI was 44.3 (± 29.4) in the in utero SGA group, which was significantly higher than that in the normal group (8.8 (± 10.0); p < 0.01) during pregnancy. There was a significant correlation between the PI and z score for estimated fetal weight (EFW) (r = -0.55; p < 0.01). Moreover, a significant positive correlation was observed between the PI and the z score of birth weight (r = -0.39; p < 0.01). Pathological ischemia findings of the placenta were identified in 67% of the HT score 3 group, representing 6 of the 9 patients, and in 20% of the HT score 1 group, representing only 3 of the 15 patients. Conclusions Placental hardness, as determined by elastography, correlates with both lower estimated fetal body weight and birth weight. These results suggest that ultrasound elastography in the placenta may be an additional marker of intrauterine fetal well-being.

Clinical efficacy of a combination of Percoll continuous density gradient and swim-up techniques for semen processing in HIV-1 serodiscordant couples.

To evaluate the clinical efficacy of a procedure comprising a combination of Percoll continuous density gradient and modified swim-up techniques for the removal of human immunodeficiency virus type 1 (HIV-1) from the semen of HIV-1 infected males, a total of 129 couples with an HIV-1 positive male partner and an HIV-1 negative female partner (serodiscordant couples) who were treated at Keio University Hospital between January 2002 and April 2012 were examined. A total of 183 ejaculates from 129 HIV-1 infected males were processed. After swim-up, we successfully collected motile sperms at a recovery rate as high as 100.0% in cases of normozoospermia (126/126 ejaculates), oligozoospermia (6/6), and asthenozoospermia (36/36). The recovery rate of oligoasthenozoospermia was 86.7% (13/15). In processed semen only four ejaculates (4/181:2.2%) showed viral nucleotide sequences consistent with those in the blood of the infected males. After using these sperms, no horizontal infections of the female patients and no vertical infections of the newborns were observed. Furthermore, no obvious adverse effects were observed in the offspring. This protocol allowed us to collect HIV-1 negative motile sperms at a high rate, even in male factor cases. We concluded that our protocol is clinically effective both for decreasing HIV-1 infections and for yielding a healthy child.