Identifying prognostic factors in Japanese women with pseudomyxoma peritonei: a retrospective clinico-pathological study of the Tohoku Gynecologic Cancer Unit.

Pseudomyxoma peritonei (PMP) is a rare condition of mucinous ascites associated mainly with mucinous tumors of appendix or ovary. PMP often recurs after treatment and may eventually cause death by abdominal visceral dysfunction via compression with mucinous ascites. Although radical peritonectomy and hyperthermic intra-peritoneal chemotherapy are becoming popular globally, the optimal treatment of PMP has not been established in Japan. We conducted a retrospective multicenter study to clarify the optimal treatment and the prognostic factors of PMP. A total of 23 patients with PMP were analyzed in the Tohoku Gynecologic Cancer Unit (TGCU). Clinical and follow-up data were retrieved and a central pathology review was performed. The median follow-up period was 46 months. Eleven patients underwent complete resection. There were 7 deaths out of 13 recurrences/progressions in this period. All the recurrence/progression was confined to the abdomen. Unexpectedly, neither radical peritonectomy nor hyperthermic intra-peritoneal chemotherapy had been performed, indicating that both radical peritonectomy and hyperthermic intra-peritoneal chemotherapy are not yet popular in Japan. The medians of overall survival and disease-free period were 166 months and 30 months, respectively. Univariate and multivariate analyses revealed that the only prognostic factor was macroscopic residual tumor (P=0.022). Although chemotherapy was not a prognostic factor (P=0.16), those who received intra-peritoneal chemotherapy tended to have a better prognosis than those who received systemic or no chemotherapy (P=0.064). In conclusion, the macroscopic residual tumor is an important prognostic factor in Japanese patients with PMP.

Distinctive ultrasonographic finding of complete uterine rupture in early mid-trimester.

In many cases of uterine rupture, diagnosis is often impossible when characteristic clinical symptoms are absent. We encountered a case of suspected peritoneal pregnancy in which we were able to make a definitive diagnosis by ultrasonography of complete uterine rupture in early mid-trimester. The possibly distinctive finding is the high-echo area that extends from the endometrium to the uterine serosa. This contiguous, highly echogenic finding should be recognized as characteristic of complete rupture of the uterus.

Paracrine regulation of the resumption of oocyte meiosis by endothelin-1.

Mammalian oocytes remain dormant in the diplotene stage of prophase I until the resumption of meiosis characterized by germinal vesicle breakdown (GVBD) following the preovulatory gonadotropin stimulation. Based on genome-wide analysis of peri-ovulatory DNA microarray to identify paracrine hormone-receptor pairs, we found increases in ovarian transcripts for endothelin-1 and endothelin receptor type A (EDNRA) in response to the preovulatory luteinizing hormone (LH)/human chorionic gonadotropin (hCG) stimulation. Immunohistochemical analyses demonstrated localization of EDNRA in granulosa and cumulus cells. In cultured preovulatory follicles, treatment with endothelin-1 promoted oocyte GVBD. The stimulatory effect of endothelin-1 was blocked by cotreatment with antagonists for the type A, but not related type B, receptor. The stimulatory effect of hCG on GVBD was partially blocked by the same antagonist. The endothelin-1 promotion of GVBD was found to be mediated by the MAPK/ERK pathway but not by the inhibitory G protein. Studies using cumulus-oocyte complexes and denuded oocytes demonstrated that the endothelin-1 actions are mediated by cumulus cells. Furthermore, intrabursal administration with endothelin-1 induced oocyte GVBD in preovulatory follicles. Our findings demonstrate a paracrine role of endothelin-1 in the induction of the resumption of meiosis and provide further understanding on the molecular mechanisms underlying the nuclear maturation of oocytes induced by the preovulatory LH surge.

Histopathological prognostic factors predicting para-aortic lymph node metastasis in patients with endometrioid uterine cancer.

INTRODUCTION: The purpose of this study was to determine histopathological factors for para-aortic lymph node (PALN) metastasis in patients with endometrioid uterine cancer. METHODS: A total of 355 patients (Stage I, n=269; II, n=24; and III, n=62) (FIGO 2009) underwent primary radical surgery including complete systematic pelvic lymph node (PLN) and PALN dissection in Tohoku Gynecologic Cancer Unit (TGCU) between 1993 and 2004. Logistic regression analysis was used to determine the independent prognostic factors for PALN metastasis. RESULTS: Multivariate analysis revealed that PLN metastasis (p<0.0001) and ovarian metastasis (p=0.0080) related with PALN metastasis. Moreover, among the sites of PLN metastases, obturator lymph node (LN) [risk ratio (RR): 16.9, 95% confidence interval (CI): 4.3-66.4, p<0.0001] and common iliac LN (RR: 7.1, 95% CI: 1.1-44.5, p=0.0375) related with PALN metastases. In detection of PALN metastasis, combination of obturator LN and/or common iliac LN and/or ovarian metastasis (A) revealed 75.9% sensitivity (22/29) and 97.8% negative predictive value (NPV) (304/311). However, by combination of obturator LN metastasis and/or common iliac LN metastasis and/or grade 3 and/or deep myometrial invasion (B), the detection of PALN metastasis was 100.0% sensitivity (29/29) and 100.0% NPV (198/198). Also, 55.8% (198/355) of patients could have avoided PALN dissection by combination B. CONCLUSIONS: These results suggest that PALN dissection is necessary when combination B is positive by pre- and intra-operative assessments. Further prospective randomized controlled studies need to be conducted in a larger patient population to establish the strategy for detecting PALN metastasis utilizing pre-/intra-operative assessments.

Time-related changes of cervical length as a predictor of labor onset within one week using transvaginal ultrasonography.

OBJECTIVE: To assess the value of transvaginal cervical length (CL) measurement and funneling of the internal cervical os to predict the onset of spontaneous labor within one week. DESIGN: Observational prospective study. POPULATION: A total of 234 women who delivered after spontaneous onset of labor. METHODS: CL was measured and the presence of funneling of the internal cervical os was recorded by transvaginal ultrasound examination. MAIN OUTCOME MEASURES: CL changes from 37 to 40 weeks of gestation as a one-point measurement, as well as the long-term (16-21 weeks to term or 37-40 weeks) and short-term (weekly from 37 to 40 weeks) differences in CL. RESULTS: The one-point measurement of CL, with an optimum cut-off point of 25 mm [odds ratio (OR) 3.19; 95% confidence interval (CI) 2.01-5.05] and a decreased long-term CL (cervical shortening of 55% as an optimum cut-off point (OR 3.75; 95% CI 1.76-8.00) may predict the onset of spontaneous labor within one week. The likelihood of spontaneous labor within one week is high when funneling of the internal cervical os is present (OR 2.68; 95% CI 1.71-4.19). CONCLUSIONS: Transvaginal CL measurement and observation of funneling of the internal cervical os are useful predictors of the onset of spontaneous labor.

Mayer-Rokitansky-Kustner Hauser syndrome complicated by either uterine leiomyoma or ovarian tumor.

A 50-year-old Japanese woman with Mayer-Rokitansky-Kustner-Hauser syndrome and two pelvic tumors underwent laparotomy. Laparotomy revealed that no torsion of the right ovarian tumor had occurred, with uterine leiomyoma originating from the right side of a rudimentary uterus. Histopathological examination demonstrated leiomyoma of the rudimentary uterus with positive staining for estrogen and progesterone receptors, and mucinous cyst adenoma of the right ovary. Uterine leiomyoma is rare in this syndrome and the present report represents the first published case complicated by ovarian tumor.

Scleroderma-like cutaneous lesions induced by paclitaxel and carboplatin for ovarian carcinoma, not a single course of carboplatin, but re-induced and worsened by previously administrated paclitaxel.

Scleroderma-like cutaneous lesion as an adverse event from paclitaxel and carboplatin has been reported. No report shows the occurrence of scleroderma-like cutaneous lesions from a single course of carboplatin. The patient is a 67-year-old female, administered paclitaxel and carboplatin as neoadjuvant chemotherapy. Following four courses, scleroderma-like cutaneous lesions were demonstrated. Skin biopsy corresponded to histopathological findings of scleroderma. Immunological investigation shows only antinuclear antibodies are positive. The characteristic Raynaud's phenomenon of scleroderma and hemorrhagic spots on the cuticles were not found. Postoperatively, a single course of carboplatin treatment was given. Scleroderma-like cutaneous lesions re-induced and worsened. This is the first report detailing scleroderma-like cutaneous lesions induced by previously administrated paclitaxel that worsened by carboplatin.

Evaluation of possible criteria for elective single embryo transfer.

Purpose: A major problem of assisted reproductive technology (ART) is multiple gestation, which impacts neonatal and perinatal medicine. The literature contains a number of reports that elective single embryo transfer (eSET) is effective for the control of multiple pregnancies; however, to date, uniform criteria have not been established. Methods: Using logistic regression analysis based on the results of ART in our department from January 2005 to July 2006, our eSET criteria were established. We conducted a comparative study of the clinical pregnancy rate, multiple gestation rate, and delivery rate before and after eSET (before-eSET and after-eSET groups, respectively). Results: As a result of the analysis, our eSET criteria included all three of the following: (A) patient age ≤37, (B) previous IVF/ICSI trials ≤5, and (C) acquisition of two or more good-quality embryos. Based on our criteria, the after-eSET group was not found to have a decrease in the pregnancy rate; however, the multiple gestation rate decreased as compared to the before-eSET group. In addition, as a result of various evaluations of the eSET group, interesting findings were revealed. Conclusions: In the after-eSET group, our eSET criteria achieved a decrease in the multiple pregnancy rate without a decrease in the pregnancy rate.

Completion of Meiosis I of preovulatory oocytes and facilitation of preimplantation embryo development by glial cell line-derived neurotrophic factor.

Optimal maturation of oocytes and successful development of preimplantation embryos is essential for reproduction. We performed DNA microarray analyses of ovarian transcripts and identified glial cell line-derived neurotrophic factor (GDNF) secreted by cumulus, granulosa, and theca cells as an ovarian factor stimulated by the preovulatory LH/hCG surge. Treatment of cumulus-oocyte complexes with GDNF enhanced first polar body extrusion with increase in cyclin B1 synthesis and the GDNF actions are likely mediated by its receptor GDNF family receptor-alpha1 (GFRA1) and a co-receptor ret proto-oncogene (Ret), both expressed in oocytes. However, treatment with GDNF did not affect germinal vesicle breakdown and cytoplasmic maturation of oocytes. During the preimplantation stages, GDNF was expressed in pregnant oviducts and uteri, whereas GFRA1 and Ret were expressed in embryos throughout early development with an increase after the early blastocyst stage. In blastocysts, both GDNF and GFRA1 were exclusively localized in trophectoderm cells, whereas Ret was detected in both cell lineages. Treatment with GDNF promoted the development of two-cell-stage embryos into blastocysts showing increased cell proliferation and decreased apoptosis mainly in trophectoderm cells. Our findings suggest potential paracrine roles of GDNF in the promotion of completion of meiosis I and the development of early embryos.

Kit ligand promotes first polar body extrusion of mouse preovulatory oocytes.

BACKGROUND: Shortly after stimulation by the preovulatory surge of luteinizing hormone (LH), oocytes arrested at the late prophase I resume meiosis characterized by germinal vesicle breakdown (GVBD), chromosome condensation, and extrusion of the first polar body in preparation for fertilization and early embryonic development. However, oocytes express few or no LH receptors and are insensitive to direct LH stimulation. Thus, factors released by granulosa or theca cells expect to convey the LH stimuli to oocytes. To identify candidate ligand-receptor pairs potentially involved in the process of oocyte maturation, we performed DNA microarray analyses of ovarian transcripts in mice and identified Kit ligand (Kitl) as an ovarian factor stimulated by the LH/hCG surge. The purpose of this study is to investigate the roles of KITL in the nuclear and cytoplasmic maturation of preovulatory mouse oocytes. METHODS: The levels of Kitl and c-kit transcripts in mouse ovaries and isolated ovarian cells were determined by real-time RT-PCR, while expression of KITL protein was examined by immunohistochemistry. Follicle culture, cumulus-oocyte complexes (COC) and denuded oocytes culture were used to evaluate the effect of KITL on mouse oocyte nuclear maturation. To assess the effect of KITL treatment on the cytoplasmic maturation of preovulatory oocytes, we performed in vitro maturation of oocytes followed by in vitro fertilization. RESULTS: Major increase of Kitl transcripts in granulosa cells and mouse ovaries, and predominant expression of c-kit in preovulatory oocytes were identified by real-time RT-PCR. Predominant expression of KITL protein was found in granulosa cells of preovulatory and small antral follicles at 4 h after hCG treatment. In vitro cultures demonstrated that treatment with KITL enhanced first polar body extrusion in a dose-dependent manner. Moreover, treatment of COC with KITL enhanced first polar body extrusion with increase in cyclin B1 synthesis which is important for the progression of meiotic maturation after GVBD. In contrast, treatment of cultured preovulatory follicles with KITL did not affect GVBD and KITL has no effect on cytoplasmic maturation of preovulatory oocytes. CONCLUSION: Our findings suggest potential paracrine roles of KITL in the nuclear maturation of preovulatory oocytes by promoting first polar body extrusion.

Endometrioid uterine cancer: histopathological risk factors of local and distant recurrence.

OBJECTIVES: To determine the relationship between histopathological prognostic factors and sites of initial recurrence in endometrioid uterine cancer. METHODS: A total of 355 patients (Stage I, n=227; II, n=38; III, n=90) underwent primary radical surgery including complete systematic pelvic lymph node (PLN) and para-aortic lymph node (PALN) adenectomy followed by adjuvant chemotherapy who were at risk for recurrence. Relapse-free survival (RFS) and disease-related survival (DRS) were analyzed using the log-rank testing. Multivariate Cox regression analysis and logistic regression analysis were used to determine and estimate independent prognostic factors. RESULTS: Lymph-vascular space invasion (LVSI), architectural grade (AG), myometrial invasion, and PLN metastasis (PLNM) were identified as independent prognostic factors for RFS. AG (p=0.0043) related with local recurrence. Among patients who received adjuvant chemotherapy, patients with G3 tumor had higher ratio of recurrence (16/45) compared with G1/2 tumor (11/102) (p=0.0004). Meanwhile, PLNM related with distant recurrence (p=0.0008). There was a statistically significant difference in RFS according to the number of positive PLN sites (group 0: n=313, 1: n=16, > or =2: n=26), five-year RFS in each group was 91.9%, 81.3%, and 41.2%, respectively. CONCLUSIONS: Sites of initial recurrence were related with AG and PLNM in patients with endometrioid uterine cancer. Current chemotherapy alone may not be an effective adjuvant therapy to prevent recurrence in patients with G3 tumor and > or =2 positive PLN sites. Prospective clinical trial needs to be conducted to establish the strategy of adjuvant therapy with these patients.

Leptin and ObRa/MEK signalling in mouse oocyte maturation and preimplantation embryo development.

Recent studies indicate that LH stimulates production of ovarian paracrine factors that induce meiosis of the oocyte. DNA microarray analyses of ovarian transcripts were performed in mice and major increases of a short isoform of leptin receptor, ObRa, were identified by the preovulatory LH/human chorionic gonadotrophin (HCG) surge. In oocytes, the level of ObRa transcripts was increased shortly after HCG stimulation, whereas the level of ObRb transcripts was not changed. Leptin was produced by cumulus, granulosa, theca and interstitial cells of ovaries and its transcript level was not regulated during gonadotrophin treatment. Treatment with leptin promoted germinal vesicle breakdown (GVBD) in oocytes within preovulatory follicles, and enhance first polar body extrusion in both cumulus-oocyte complexes and denuded oocytes. The leptin-promoted GVBD and first polar body extrusion were blocked by a mitogen-activated protein kinase extracellular signal regulated kinase kinases (MEK)1/2 inhibitor, U0126, but not its inactive analogue U0124. Furthermore, leptin promoted fertilization of oocytes and the in-vitro development of zygotes to preimplantation embryos. These findings suggest paracrine roles of leptin in the enhancement of nuclear maturation of oocytes through MEK1/2 signalling, and in the promotion of cytoplasmic maturation essential for successful oocyte development to the preimplantation embryos.

Role of complete para-aortic lymphadenectomy in endometrial cancer.

PURPOSE OF REVIEW: Many reports suggest the significance of pelvic lymph-node (PLN) adenectomy in patients with endometrial cancer. However, among these, there is controversy regarding not only what type of patients should have lymphadenectomy performed, but also what extent lymphadenectomy should be performed. RECENT FINDINGS: It has been reported that PLN adenectomy has therapeutic significance in stage I grade 3 and more advanced endometrioid uterine cancer. However, the effects of para-aortic lymph-node adenectomy on its prognostic benefit have not been discussed. SUMMARY: Patients with low-risk disease might not benefit from PLN adenectomy. However, PLN adenectomy might still have merit in low-risk patients, as there are inaccuracies of preoperative and intraoperative assessments. A complete lymphadenectomy is safe with minimum complications. At this point, hysterectomy and bilateral salpingo-oophorectomy with complete PLN adenectomy as the standard surgical procedure for endometrial cancer is thought to be reasonable. At present, the addition of p-aortic lymph-node adenectomy is regarded as an investigated protocol for endometrial cancer. However, p-aortic lymph-node adenectomy may have a therapeutic role for stage IIIC patients. Prospective randomized controlled trial composed of intermediate/high-risk patients should be conducted to clearly demonstrate prognostic improvement by p-aortic lymph-node adenectomy itself.

Fetal preload index of the inferior vena cava and neonatal outcome of congenital diaphragmatic hernia.

PURPOSE: The purpose of this study was to assess prognostic factors in patients with congenital diaphragmatic hernia (CDH). METHODS: Thirteen patients with CDH diagnosed antenatally and delivered in our hospital between 1995 and 2006 were retrospectively studied. Assessments of sonographic examinations included gestational age at time of diagnosis; the ultrasonographic parameters [amniotic fluid index, cardiothoracic area ratio, and the lung-thoracic transverse area ratio (LTR)]; and the incidence of polyhydramnios, intrauterine growth retardation, and hydrops. Doppler velocimetry measurements comprised the resistance index of the umbilical artery, the resistance index of the midcerebral artery, the maximal velocity of the descending aorta, and the preload index of the inferior vena cava (IVCPLI). Results were expressed as the mean ± standard deviation. The features of survivors and nonsurvivors were compared. RESULTS: Six fetuses were survivors and seven were nonsurvivors. The mean LTR value tended to be markedly low in both groups (23.8 ± 16.4 vs 12.1 ± 1.1). In Doppler analysis, the mean IVCPLI value in survivors was significantly lower than that in nonsurvivors (0.34 ± 0.08 vs 0.52 ± 0.14, P = 0.01). CONCLUSION: We concluded that fetal IVCPLI might be a good predictor of the outcome in patients with CDH.

Regulation of preimplantation embryo development by brain-derived neurotrophic factor.

Hormonal factors secreted by embryos and reproductive tracts are important for successful development of preimplantation embryos. We found expression of brain-derived neurotrophic factor (BDNF) and neurotrophin-4/5 (NT-4/5) transcripts at its highest levels in the blastocyst stages. The transcripts for their receptor, TrkB, were detectable throughout the early embryonic stages with an increase after the early blastocyst stage. Both BDNF and TrkB are expressed in trophectoderm cells, whereas ligand-binding studies indicated specific binding of BDNF to trophectoderm cells. Furthermore, BDNF and NT-4/5 were produced in pregnant oviducts and uteri. Treatment with BDNF promoted the development of two-cell-stage embryos into blastocysts showing increased proliferation and decreased apoptosis. The effects of BDNF were blocked by the TrkB ectodomain or a Trk receptor inhibitor, K252a. Studies using specific inhibitors demonstrated the roles of the PI3K, but not the ERK, pathway in mediating BDNF actions. Under high-density embryo cultures, treatment with the TrkB ectodomain or K252a alone also inhibited embryonic development and survival, suggesting potential autocrine actions of BDNF produced by the embryo. In vivo experiments further demonstrated that K252a treatment suppressed early embryo development by inhibiting blastocyst cell numbers, and increasing blastocyst apoptosis. Our findings suggested that BDNF signaling plays important paracrine roles during blastocyst development by promoting the development of preimplantation embryos.

A phase II multicenter trial of concurrent chemoradiotherapy with weekly nedaplatin in advanced uterine cervical carcinoma: Tohoku Gynecologic Cancer Unit Study.

The purpose of this study was to evaluate the effectiveness and safety of concurrent chemoradiotherapy using weekly nedaplatin for the treatment of locally advanced squamous cell carcinoma of the uterine cervix. Nedaplatin at 30 mg/m(2) was administered weekly 6 times with a concurrent external beam and intracavity radiotherapy. External beam radiation was delivered with a fraction dose of 2 Gy per day for 5 days a week during a 5-week period and intracavitary brachytherapy, of which the fraction size is 6 Gy to point A, was given once a week for a total of 4 times using a remote after-loading system. Forty-five patients were enrolled in this trial between April 2003 and December 2006. Of the 45 patients, 40 (88.9%) completed the scheduled treatment and were evaluated for efficacy and safety. Of these, 4 were stage Ib2, 12 were stage IIb, 18 were stage IIIb and 6 were stage IVa. The age distribution ranged from 27 to 79 years with a median age of 58. The 40 patients achieved an objective response, 36 (90%) a complete response and 4 (10%) a partial response. At a median follow-up of 29 months (range, 8-52), the 3-year progression-free and overall survival were 58.7% (95% confidence interval, 42-75%) and 78.0% (95% confidence interval, 56-90.0%), respectively. Acute toxicities were transient and rendered non-lethal. Of the 45 patients enrolled for the trial, only 3 (6.7%) had grade 4 leukopenia and neutropenia, respectively. Grade 3 diarrhea and nausea/ vomiting were observed in 2 (4.4%) and 1 (2.2%), respectively. These results indicate that weekly nedaplatin of 30 mg/m(2) with concurrent radiotherapy is an effective and well-tolerated regimen for advanced squamous cell carcinoma of the uterine cervix.

Para-aortic lymphadenectomy may improve disease-related survival in patients with multipositive pelvic lymph node stage IIIc endometrial cancer.

OBJECTIVE: The purpose of this study was to determine whether para-aortic lymphadenectomy improves disease-related survival (DRS) in stage IIIc endometrial cancer. METHODS: A total of 63 patients with stage IIIc endometrial carcinoma underwent primary radical surgery in the Tohoku Gynecologic Cancer Unit from 1993 to 2004. All patients had modified radical hysterectomy, bilateral salpingo-oophorectomy, systemic pelvic lymph node (PLN) adenectomy, and with or without para-aortic lymph node (PAN) adenectomy, followed by adjuvant chemotherapy. DRS was analyzed using Kaplan-Meier curves and the log-rank test. Independent prognostic factors were determined by multivariate Cox regression analysis using a forward stepwise selection. RESULTS: There were no statistical differences in age distribution and histopathological prognostic factors between PLN adenectomy group (n=25) and PLN+PAN adenectomy group (n=38). On univariate analysis, architectural grade (p=0.026), peritoneal cytology (p=0.033), and the number of PLN positive sites (/=2) (p=0.010) were related to poor DRS. On multivariate Cox regression analysis, the number of positive PLN sites was related to DRS (p=0.040). In positive PLN>/=2 sites group (n=33), PAN adenectomy significantly improved DRS compared to PLN adenectomy alone (p=0.011). The incidence of initial PAN recurrence was higher in the PLN adenectomy group (6/25) than in the PLN+PAN adenectomy group (1/38) (p=0.013, Odds Ratio=11.68). CONCLUSIONS: The number of positive PLN site is an independent prognostic factor in stage IIIc endometrial cancer. PAN adenectomy decreased the incidence of PAN recurrence and may improve DRS in patients with >/=2 positive PLN sites. A large prospective clinical trial needs to be conducted to establish the strategy of PAN adenectomy before or intra-operative treatment.

Roles of forkhead transcription factor Foxc2 (MFH-1) and endothelin receptor A in cardiovascular morphogenesis.

OBJECTIVE: Foxc2/MFH-1 is a member of the forkhead family of transcription factors and Foxc2-deficient mice exhibit aortic arch anomalies (type B interruption of the aortic arch). Endothelin receptor type-A (ETA) is one of the two known endothelin receptors that belong to the G-protein-coupled receptor family. ETA-deficient mice show defects in the great arteries, primarily type B interruption of the aortic arch. Based on similar phenotypes in the cardiovascular system of Foxc2- and ETA-deficient mice, we investigated whether Foxc2 and ETA have a close relationship in aortic arch patterning. METHODS: The Foxc2 and ETA homozygotes were obtained by crossing the Foxc2 and ETA heterozygotes, respectively. The double Foxc2/ETA homozygotes were obtained by crossing the double Foxc2/ETA heterozygotes. RESULTS: We investigated the expression of ETA in Foxc2-null mice and the expression of Foxc2 in ETA-null mice and found that the absence of either Foxc2 or ETA had no effect on the expression of the other. Next, we analyzed mice lacking both Foxc2 and ETA to examine the relationship between Foxc2 and ETA on aortic arch patterning in vivo. We found that the majority of Foxc2/ETA double-mutant embryos died around 11.5 dpc and that all surviving mice had persistent truncus arteriosus. CONCLUSIONS: The results suggest that Foxc2- and ETA-expressing cells additively form the aorticopulmonary septum.

Gonadotropin-releasing hormone I analog acts as an antiapoptotic factor in mouse blastocysts.

Both GnRH-I and its receptor (GnRHR)-I have been shown to be expressed in the mammalian preimplantation embryo. In this study, we investigated the molecular mechanisms of GnRH-I in the regulation of early embryonic development in mouse. We found that GnRH-I and GnRHR-I mRNAs were detectable throughout early embryonic stages and that expression levels of both increased significantly after the early blastocyst stage. In blastocysts, GnRH-I and GnRHR-I expression was detected in both inner cell mass and trophectoderm cells. The pregnant uterus also expressed both genes, suggesting that preimplantation embryos could be affected by GnRH through both paracrine and autocrine signaling. Treatment with GnRH-I agonist, buserelin, promoted development of two-cell-stage embryos to the expanded and hatched blastocyst stages and inhibited apoptosis in a dose-dependent manner. In contrast, treatment with GnRH-I antagonist, ganirelix acetate, inhibited development of preimplantation embryos beyond the expanded blastocyst stage and induced apoptosis; both effects could be reversed by cotreatment with GnRH-I agonist. GnRH-I antagonist-induced cell death was mediated by disruption of mitochondrial function, release of cytochrome c, and activation of caspase-3. Furthermore, treatment with GnRH-I antagonist decreased expression of two antiapoptotic growth factors, epidermal growth factor and IGF-II, in blastocysts. These results indicate that GnRH-I, acting as an antiapoptotic factor, is an important growth factor in development of mouse blastocysts.

Right ventricular pre-ejection myocardial velocity and myocardial acceleration in normal fetuses assessed by Doppler tissue imaging.

Myocardial acceleration during isovolumic contraction obtained from Doppler tissue imaging has been introduced as an index of right ventricular contractile function that is unaffected by the shape of the ventricle and loading conditions, but normal value of myocardial acceleration during isovolumic contraction and the effect of aging on the index are not known in normal fetuses. We studied 61 normal fetuses aged 20 to 39 weeks (29.8 +/- 5.1 weeks). Fetuses were divided into 4 age groups: 20 to 24 weeks (n = 11); 25 to 29 weeks (n = 20); 30 to 34 weeks (n = 20); and 35 to 39 weeks (n = 10). Using Doppler tissue imaging, peak pre-ejection myocardial velocity was measured at the base of right ventricular free wall from 4-chamber view. Myocardial acceleration was calculated by dividing pre-ejection velocity by the time interval from onset of the pre-ejection myocardial velocity to the time at peak velocity of this wave. The mean pre-ejection myocardial velocity was 5.0 +/- 1.1 cm/s. There was a stepwise increase in the pre-ejection myocardial velocity from the fetuses of 20 to 24 weeks to the fetuses of 35 to 39 weeks. The mean myocardial acceleration was 160 +/- 30 cm/s 2 . The mean myocardial acceleration did not differ between the fetuses aged 20 to 24 weeks (139 +/- 13 cm/s 2 ) and the fetuses aged 25 to 29 weeks (143 +/- 21 cm/s 2 ), but after 30 weeks increased with gestational age. For the total group combining the 4 different gestational age groups, the pre-ejection myocardial velocity and myocardial acceleration correlated with gestational age ( r = 0.85 and 0.75). This study demonstrated the gestational age-related changes in pre-ejection myocardial velocity and myocardial acceleration. The load-independent index of contractility, myocardial acceleration, increased mainly after 30 weeks' gestation.