Expression patterns of emmprin and monocarboxylate transporter-1 in ovarian epithelial tumors.

Emmprin is a transmembrane glycoprotein known as a matrix metalloproteinase inducer and is highly up-regulated in malignant cancer cells. The monocarboxylate transporters (MCTs) are responsible for H(+)-linked transport of monocarboxylates across the cell membrane. It was recently demonstrated that proper plasma membrane localization and activity of MCTs require the presence of emmprin as a chaperone and that MCT-1 also acts as chaperone for emmprin. The objectives of this study were to clarify emmprin and MCT-1 expression patterns in ovarian epithelial tumors and to elucidate the clinicopathological significance of co-localization of the two molecules. Immunohistochemical analysis of 205 epithelial tumors indicated that emmprin is always localized in cell membranes but its distribution differs according to tumor type: in lateral membranes in 89 % of adenomas, in lateral and basal membranes in 76 % of borderline tumors, and in membranes surrounding the entire cell in 98 % of carcinomas. Most carcinomas in situ also showed a lateral and basal expression pattern. In only 21 % of the carcinomas, the cells expressing membranous MCT-1 showed co-localized emmprin expression. Poor co-localization of the two molecules was more frequently found in serous carcinomas. However, the overall survival was not significantly different for the good and poor co-localization carcinoma groups. These findings indicate that the emmprin expression pattern might discriminate between invasive carcinomas and borderline tumors including carcinoma in situ. Moreover, there may be an as yet unidentified regulatory mechanism(s), for localization of MCT-1 and emmprin in cell membranes in vivo.

Magnesium sulfate as a second-line tocolytic agent for preterm labor: a randomized controlled trial in Kyushu Island.

OBJECTIVES: We evaluated the efficacy of magnesium sulfate as a second-line tocolysis for 48 hours. MATERIALS AND METHODS: A multi-institutional, simple 2-arm randomized controlled trial was performed. Forty-five women at 22 to 34 weeks of gestation were eligible, whose ritodrine did not sufficiently inhibit uterine contractions. After excluding 12 women, 33 were randomly assigned to either magnesium alone or combination (ritodrine and magnesium). The treatment was determined as effective if the frequency of uterine contraction was reduced by 30% at 48 hours of the treatment. RESULTS: After magnesium sulfate infusion, 90% prolonged their pregnancy for >48 hours. Combination therapy was effective in 95% (18/19), which was significantly higher than 50% (7/14) for magnesium alone. CONCLUSION: This randomized trial revealed that combination therapy significantly reduced uterine contractions, suggesting that adjuvant magnesium with ritodrine is recommended, rather than changing into magnesium alone, when uterine contractions are intractable with ritodrine infusion.

Metronomic doxifluridine chemotherapy combined with the anti-angiogenic agent TNP-470 inhibits the growth of human uterine carcinosarcoma xenografts.

Uterine carcinosarcoma is a highly aggressive gynecological neoplasm that responds poorly to conventional chemotherapy and radiotherapy. Metronomic chemotherapy is accepted as a new approach for cancer treatment, and its underlying mechanism seems to involve the suppression of angiogenesis. However, the efficacy of metronomic and anti-angiogenic therapies against uterine carcinosarcoma is unknown. The anti-angiogenic effect of doxifluridine was assessed in vitro using human umbilical vein endothelial cells (HUVEC) co-cultured with FU-MMT-1 human uterine carcinosarcoma cells. The antitumor and anti-angiogenic effects of metronomic doxifluridine (delivered via oral gavage) in combination with TNP-470 were evaluated in vivo. Tumor vascularity was assessed by contrast-enhanced color Doppler ultrasound, laser Doppler and microvessel density staining. Doxifluridine suppressed tube formation of HUVEC and vascular endothelial growth factor production by FU-MMT-1 cells. Metronomic doxifluridine alone significantly suppressed tumor growth compared with the untreated (control) group, while metronomic doxifluridine in combination with TNP-470 significantly inhibited tumor growth compared with each treatment alone. A significant reduction of intratumoral vascularity was observed in FU-MMT-1 xenografts following treatment with metronomic doxifluridine in combination with TNP-470, as compared with each treatment alone. Intestinal bleeding was only observed when the maximum tolerated dose of doxifluridine was administered in combination with TNP-470. Metronomic doxifluridine chemotherapy in combination with TNP-470 might be effective for uterine carcinosarcoma without marked toxicity, possibly acting via its potent anti-angiogenic effects. Clinical studies are needed to evaluate the safety and efficacy of this treatment in humans.

Clinical usefulness of contrast-enhanced color Doppler ultrasonography in invasive and noninvasive gestational trophoblastic diseases: a preliminary study.

OBJECTIVE: To examine the usefulness of contrast-enhanced color Doppler ultrasonography (CDU) in differentiating between invasive and noninvasive gestational trophoblastic disease (GTD). STUDY DESIGN: In 23 patients with findings suggestive of GTD by transvaginal gray-scale ultrasonography, the presence or absence of blood flow within uterine lesions was assessed by contrast-enhanced CDU using Levovist (Schering, Berlin, Germany) microbubble contrast agent. Intratumoral blood flow waveforms were analyzed using resistance indices. Tumor size in each invasive or malignant GTD was assessed by magnetic resonance imaging. RESULTS: Intratumoral blood flow was detected in all invasive or malignant GTDs (7/7: 5 invasive moles, 1 choriocarcinoma and 1 placental site trophoblastic tumor), whereas it was not seen in any noninvasive GTD (0/16:10 complete moles, 5 partial moles and 1 exaggerated placental site) (p <0.0001). A marked increase in uterine vascularity was thus shown in all invasive or malignant GTDs following enhancement. In small invasive moles (<2 cm) in the uterine myometrium, color flow was remarkably increased by contrast-enhanced CDU. Intratumoral blood flow waveforms showed low resistance indices in all invasive and malignant GTDs. CONCLUSION: Contrast-enhanced CDU may be useful in differentiating invasive or malignant GTDs from noninvasive GTDs. By enhancing color flow, this minimally invasive approach may be helpful for detecting small invasive GTD lesions within the uterine myometrium.

Metronomic irinotecan chemotherapy combined with ultrasound irradiation for a human uterine sarcoma xenograft.

Metronomic chemotherapy is the frequent administration of low doses of chemotherapeutic agents targeting tumor-associated endothelial cells. We examined the efficacy of metronomic irinotecan combined with low-intensity ultrasound (US) in human uterine sarcoma and evaluated its antiangiogenesis mechanism by measuring the circulating endothelial progenitor cells (CEP), a surrogate marker of angiogenesis. A human uterine sarcoma cell line, FU-MMT-3, was used in the present study because this tumor is one of the most malignant neoplasms of human solid tumors and it also has a high angiogenesis property. The combination of low-dose irinotecan and US irradiation significantly inhibited the tube formation of HUVEC and vascular endothelial growth factor expression of tumor cells in vitro. The FU-MMT-3 xenografts in nude mice were treated using US at a low intensity (2.0 w/cm(2), 1 MHz) for 4 min three times per week each after the intraperitoneal administration of irinotecan; this treatment was continued for 5 weeks. The tumor vascularity was assessed by contrast-enhanced color Doppler US in real time. The combination treatment significantly inhibited the mobilization of CEP and intratumoral vascularity compared with the control. This combination therapy showed a significant reduction in tumor volume, resulting in a significant prolongation of survival, in comparison with each treatment alone. These results suggest that the effect of metronomic chemotherapy for human uterine sarcoma was accelerated by US irradiation in vivo and this combination might therefore be potentially effective for new cancer therapy.

Apoptosis as a possible candidate mechanism for removal of tamoxifen-related endometrial cells with KRAS mutations.

BACKGROUND: Endometrial cell KRAS mutations are frequent in tamoxifen (TAM)-treated breast cancer patients. We previously demonstrated that most KRAS mutations disappeared after TAM cessation, suggesting the existence of a removal mechanism for endometrial cells with KRAS mutation. Here, the role of apoptosis in this mechanism was investigated. PATIENTS AND METHODS: DNA was extracted from frozen endometrial polyps of 31 TAM-treated breast cancer patients. Codon 12 mutations in KRAS were detected by enriched polymerase chain reaction enzyme-linked minisequence assay. Apoptosis was detected by the TdT-mediated dUTP-biotin nick end-labeling (TUNEL) method and Ki-67 expression by immunohistochemistry. Relationships between KRAS mutations, the apoptosis index, and the Ki-67 index were determined. RESULTS: KRAS mutations were observed in 9 of these patients. There was no significant relationship between the Ki-67 index and KRAS mutation. However, the apoptosis index was significantly higher in polyps with KRAS mutation (p=0.002). CONCLUSION: Apoptosis may play an important role in removing TAM treatment-related endometrial cells with KRAS mutations.

HB-EGF inhibition in combination with various anticancer agents enhances its antitumor effects in gastric cancer.

Advanced gastric cancer (GC) is one of the most lethal malignancies. Although many anticancer agents exist for the treatment of GC, its prognosis remains extremely poor. Therefore, further development of targeted therapies is required for patients with GC. To assess the role of heparin-binding epidermal growth factor-like growth factor (HB-EGF) as a target for GC therapy, the expression of EGF receptor ligands in GC cell lines, and the antitumor effects of an HB-EGF inhibitor (CRM197) as a single agent and in combination with other anticancer agents was assessed in GC cells. HB-EGF was the predominantly expressed ligand among EGF receptor ligands in all the cells. CRM197 induced significant cell apoptosis. Anticancer agents augmented the secretion of HB-EGF into the medium and simultaneously induced cell apoptosis. Combination of CRM197 with other anticancer agents significantly enhanced cell apoptosis. Additionally, co-administration of CRM197 and paclitaxel resulted in synergistic antitumor effects. These results suggested that HB-EGF is a rational target for GC therapy.

Longitudinal changes in canal length at 16-35 weeks in normal twin pregnancies and twin pregnancies with preterm labor and delivery.

AIM: To describe the longitudinal changes in canal length at 16-35 weeks' gestation in cases of twin pregnancy with preterm labor and delivery. METHODS: We studied 22 cases of twin pregnancy that were delivered at < 36 weeks and/or that underwent preterm labor requiring tocolysis. We also studied 44 cases of twin pregnancy delivered at > or = 36 weeks without tocolysis (non-tocolysis twin pregnancy). Controls were 82 cases of normal singleton pregnancy. Canal length was longitudinally measured using transvaginal ultrasonography. The observational period of 16-35 weeks was divided into 4-week periods for analysis. RESULTS: From 28 to 31 weeks onwards the canal length of non-tocolysis twin pregnancies was shorter than that of normal singleton pregnancies (P < 0.05). The canal length of twin pregnancies with preterm labor and delivery was shorter than that of non-tocolysis twin pregnancies at 16-19 weeks and decreased rapidly until 24-27 weeks (P < 0.01). CONCLUSIONS: A short canal length at 16-19 weeks followed by rapid canal length shortening in the second trimester are specific characteristics in preterm labor and delivery of twin pregnancies. Sequential measurements of canal length in the second trimester starting at < 20 weeks may be a suitable parameter to predict preterm labor and delivery in twin pregnancies.

Primary leiomyosarcoma of the fallopian tube.

Primary sarcoma of the fallopian tube is a very rare neoplasm. We report the case of a 69-year-old woman affected with leiomyosarcoma of the left fallopian tube. Her chief complaint was lower abdominal pain. The preoperative diagnosis was a left adnexal malignant tumor based on pelvic examination, abdominal computed tomography, and magnetic resonance imaging. Following a laparotomy, she was ultimately diagnosed with a FIGO IIc fallopian tube leiomyosarcoma. She was treated with total abdominal hysterectomy, bilateral salpingo-oophorectomy, pelvic lymph node dissection, partial omentectomy, and low anterior resection for rectal invasion. The patient subsequently received adjuvant chemotherapy with pirarubicin and ifosfamide. Thirty months after the first therapy, a computed tomography scan revealed metastasis of the liver, lung, and supraclavicular lymph node. The patient died of the disease 39 months after the initial treatment.

Novel chemoembolization using calcium-phosphate ceramic microsphere incorporating TNP-470, an anti-angiogenic agent.

The purpose of the present study was to develop a new method of chemoembolization to improve the therapeutic effectiveness and safety profile of cancer treatment. A chemoembolization approach was designed for human solid tumors using resorbable calcium-phosphate ceramic microspheres loaded with an agent anti-angiogenic to tumor vasculature in vivo. The human uterine sarcoma cell line FU-MMT-3 was used in this study because this tumor is aggressive and also exhibits a poor response to radiotherapy or any chemotherapy currently used. The calcium-phosphate ceramic microspheres loaded with TNP-470, an anti-angiogenic agent, were injected into FU-MMT-3 xenografts in nude mice three times per week for 8 weeks. The treatment using TNP-470-loaded microspheres suppressed tumor growth, compared to treatment with TNP-470 alone, microspheres alone, and the control. The mean tumor weight after treatment using TNP-470-loaded microspheres was significantly lower than that after treatment with microspheres alone. These ceramic microspheres were remarkably embolized in tumor microvessels as well as in the feeding arteries and a significant reduction of intratumoral vascularity was also demonstrated following treatment with TNP-470-loaded microspheres. Severe loss of body weight was not observed in any mice treated with the TNP-470-loaded microspheres, compared to treatment with TNP-470 alone. These results suggest that targeting tumor vasculature in human uterine sarcoma using calcium-phosphate microspheres might be more effective and safer than the treatment that employs anti-angiogenic agent alone. This new chemoembolization method incorporating an anti-angiogenic agent may contribute to the effective treatment of locally advanced or recurrent solid tumors.

Monitoring of endometrial K-ras mutation in tamoxifen-treated patients with breast cancer.

INTRODUCTION: A high incidence of endometrial K-ras mutations has been reported in tamoxifen (TAM)-treated patients with breast cancer. We examined the changes in the frequency of the endometrial K-ras mutations after the cessation of TAM treatment. METHODS: DNA was extracted from fresh cytological or polypectomy samples of the endometrium in 28 patients who had undergone TAM treatment of breast cancer. Mutations were detected by an enriched polymerase chain reaction-enzyme-linked minisequence assay (Sumitomo Metal Industry, Inc, Tokyo, Japan). K-ras codon 12 mutations were monitored in these 28 patients. RESULTS: An initial examination detected endometrial K-ras mutations in 13 of the 28 patients. However, repeated examinations performed after cessation of TAM treatment did not detect endometrial K-ras mutations in any of these 13 patients. No endometrial K-ras mutation has been detected in the repeated examinations performed for these patients for more than 2 years since the cessation of TAM treatment. In addition, the 15 patients who did not have endometrial K-ras mutations in the initial examination did not demonstrate them in repeat examinations. CONCLUSIONS: The cessation of TAM treatment may reduce the risk of developing endometrial cancers through K-ras mutations.

The efficacy of preoperative hormonal therapy before laparoscopic cystectomy of ovarian endometriomas.

AIM: The aim of this study was to investigate the influence of preoperative hormonal therapy before laparoscopic cystectomy of ovarian endometriomas. We identified differences in follicle loss and surgical difficulties with or without preoperative hormonal therapy. METHODS: Ninety-six patients with ovarian endometrioma underwent a laparoscopic cystectomy. Patients were divided into three groups: control group A (53 patients, 57 cysts) with no preoperative hormonal therapy; group B (34 patients, 40 cysts) who received gonadotropin-releasing hormone agonist therapy; and group C (9 patients, 11 cysts) who received danazol therapy before surgery. The medical and videotape records of all patients were retrospectively reviewed. The specimens of endometriomas were histologically evaluated. RESULTS: Mean diameters of endometriomas before hormonal therapy in groups B and C were significantly greater than those in the control group. There were no significant differences in the following: the mean diameter of removed cysts, the revised-American Society of Reproductive Medicine scores, the number of capsules containing follicle(s), and the mean number of follicles attached to a cyst. However, the number of capsules showing fibrosis significantly increased in the preoperative hormonal therapy groups (P < 0.001). Furthermore, the mean operation time of the preoperative hormonal groups was significantly longer than that of the control group (P < 0.01, P < 0.001). CONCLUSION: Our data suggested that preoperative hormonal therapy reduced the size of endometriomas. However, with similar revised-American Society of Reproductive Medicine scores, preoperative hormonal therapy did not contribute to the reduction of the loss of ovarian follicles. Fibrosis resulting from hormonal therapy appears to be responsible for these observations.

Investigation of beta(2)-adrenoceptor subtype selectivity and organ specificity for bedoradrine (KUR-1246), a novel tocolytic beta-adrenergic receptor stimulant.

OBJECTIVES: The aim of this study was to evaluate the beta-adrenergic receptor (beta-AR) selectivity, organ specificity and efficacy of delaying the onset of spontaneous delivery of bedoradrine (KUR-1246), a novel uterine relaxant. METHODS: beta-AR selectivity was evaluated in terms of the amount of cyclic adenosine monophosphate produced by bedoradrine, ritodrine and isoprenaline in Chinese hamster ovary cells expressing human beta(1)-, beta(2)-AR or beta(3)-AR. Inhibition of contractions of the atrium, trachea and proximal colon by bedoradrine were compared with those of the uterus in pregnant rats using an organ bath method. Finally, the delaying effect of bedoradrine on spontaneous labor was evaluated by an in vivo study using term pregnant rats. RESULTS: EC(50) values of bedoradrine for cyclic adenosine monophosphate production in Chinese hamster ovary cells via beta(1)-, beta(2)- and beta(3)-AR were 2400 +/- 30, 2.9 +/- 0.10 and 363 +/- 3 nmol/L, respectively, indicating that bedoradrine had 832- and 126-fold higher selectivity for beta(2)-AR than for beta(1)- and beta(3)-AR. EC(50) values of bedoradrine for the uterus, atrium, trachea and proximal colon were 1.01 +/- 0.27, 2300 +/- 356, 1610 +/- 299 and 219 +/- 23.5 nmol/L, respectively. Thus, bedoradrine was 2280-, 1590- and 217-fold more specific for the uterus than for the atrium, trachea and proximal colon, respectively. Bedoradrine delayed the spontaneous delivery of 21-day-pregnant rats in a dose-dependent manner. CONCLUSIONS: Bedoradrine is a promising drug for the treatment of preterm labor in obstetrical practice because it has better selectivity for beta(2)-AR and specificity for the uterus than currently used agents and may effectively delay spontaneous delivery.

Amphiregulin regulates the production of human chorionic gonadotropin in trophoblasts.

AIMS: The aim of this study was to investigate the significance of epidermal growth factor receptor (EGFR) ligands produced in syncytiotrophoblasts during normal pregnancy. MAIN METHODS: We examined the expression of EGFR ligands in human pregnancy by real-time PCR, and analyzed the relationship between EGFR ligands and human chorionic gonadotropin (hCG) or human placental lactogen in amniotic fluid by ELISA. In addition, we also examined the EGFR ligands in syncytiotrophoblasts and the amount of hCG secretion in JAR, JEG3 and BeWo cells in the presence of each EGFR ligand. KEY FINDINGS: In order to identify possible candidates among the EGFR ligands, we examined the predominant expression of an EGFR ligand in the chorionic villi and amniotic fluid during normal pregnancy, and analyzed the relationship between EGFR ligands and hCG in trophoblastic model cells. Amphiregulin was primarily expressed throughout human pregnancy and stimulated the secretion of hCG, indicating that amphiregulin is a key molecule among EGFR ligands. SIGNIFICANCE: Amphiregulin may play a pivotal role in the development or maturation of placenta.

Endometrial cytologic findings in tamoxifen-treated breast cancer patients.

OBJECTIVE: To describe the endometrial cytologic findings in tamoxifen (TAM)-treated patients and evaluate the clinical significance of these findings. STUDY DESIGN: A total of 1,739 endometrial cytologic samples were obtained from 203 breast cancer patients with TAM treatment using an endocyte device. Histologic examinations were recommended for the 23 endometrial cytologic samples (18 initial and 5 follow-up endometrial cytologic samples). These 23 samples were cytopathologically reviewed. RESULTS: A large nuclear dimension and anisokaryosis of the endometrial glandular cells were found in the 23 endometrial cytologic samples. All patients were amenorrheic and postmenopausal except for 3 premenopausal women with endometrial cancer. The subsequent histologic examinations showed 3 atrophic endometria, 3 cystic atrophies, 11 endometrial polyps and 6 endometrial cancers. The tumor diathesis and atypical features of cell aggregates, such as a loss of polarity and severe piling up of nuclei, were seen in the cytologic samples of the endometrial cancers but were not found in those with benign conditions. CONCLUSION: A large nuclear dimension and anisokaryosis of the glandular cells were sometimes found in endometrial cytologic samples from the TAM-treated patients. The atypical characteristics of the cell aggregates with these types of cellular atypia are useful for the cytologic assessment of malignancy.

A normal uterus communicating with a double cervix and the vagina: a müllerian anomaly without any present classification.

OBJECTIVE: To report a rare uterine anomaly consisting of a normal uterus, a double cervix, and a double vagina. DESIGN: Case report. SETTING: University hospital. PATIENT(S): A 28-year-old nulligravida patient referred for evaluation of primary infertility and a suspected müllerian anomaly. INTERVENTION(S): Clinical and surgical evaluation of the anomaly. MAIN OUTCOME MEASURE(S): Description and treatment for a rare uterine anomaly and a subsequent literature search. RESULT(S): Successful resection of vaginal septum and subsequent pregnancy. CONCLUSION(S): This extremely rare anomaly is not explained by classic embryologic teachings, and it does not fit into the classification system currently used to describe müllerian anomalies.

Amphiregulin is much more abundantly expressed than transforming growth factor-alpha and epidermal growth factor in human follicular fluid obtained from patients undergoing in vitro fertilization-embryo transfer.

OBJECTIVE: To identify the most important epidermal growth factor (EGF) receptor ligand in the LH or hCG signal pathway in human ovary. DESIGN: A retrospective clinical study. SETTING: Tertiary university hospital. PATIENT(S): Ninety-eight infertile patients who underwent IVF-embryo transfer. INTERVENTION(S): Sera and follicular fluid were collected at the time of oocyte retrieval. The levels of EGF, transforming growth factor-alpha (TGFalpha), and amphiregulin (AR) were measured in follicular fluid and sera by using ELISA. MAIN OUTCOME MEASURE(S): The relationships between the level of AR and level of hCG, fertilization rate, and embryo quality. RESULT(S): Amphiregulin was abundantly expressed in follicular fluid after hCG stimulation. Although large differences were found between AR and both EGF and TGFalpha in follicular fluid, no significant difference was detected in the levels of the three EGF receptor ligands in sera. The level of AR was inversely correlated with the fertilization rate and hCG level, whereas little significant association was observed between the level of AR and embryo quality. CONCLUSION(S): Amphiregulin was expressed most dominantly among EGF receptor ligands tested and may mediate the hCG signal in human oocyte maturation. Elaborate interaction between AR and hCG may be required for an optimal oocyte maturation.

Clinical significance of cervical length shortening before 31 weeks' gestation assessed by longitudinal observation using transvaginal ultrasonography.

AIM: To elucidate the clinical significance of progressive cervical length (CL) shortening before 31 weeks' gestation. METHODS: Transvaginal ultrasonography was used for longitudinal measurements of CL in 114 singletons. CL shortening groups were defined as having a CL of <25 mm at <26 weeks (early shortening group, 20 cases) and <31 weeks (late shortening group, 19 cases). The control group (75 cases) was defined as having a CL of >or=25 mm at <31 weeks. The CL values at 16-20, 21-25, 26-30 and 31-35 weeks, the age-related CL changes, the treatments for preterm labor, and the outcomes were compared between groups. In 78 cases with spontaneous delivery at >or=36 weeks, we investigated cervical dilatation velocity in the active phase of labor. RESULTS: At 16-20 weeks, CL values for the early group were smaller than those of the late and control groups. Rapid CL shortening occurred between 16-20 and 21-25 weeks in the early group and between 21-25 and 26-30 weeks in the late group. In the early group, all cases received cerclage and/or tocolysis or bedrest, and one case delivered prematurely. In the late group, 10 cases required tocolysis or bedrest, and one case delivered prematurely. In nulliparous women, cervical dilatation velocity in the early and late groups was more rapid than in the controls. CONCLUSIONS: CL shortening to <25 mm before 31 weeks is a risk factor for preterm delivery, as well as for preterm labor in cases who had tocolysis and bedrest, and precipitate delivery.

Modification of the endoscopic management of congenital duodenal stenosis.

This report documents a new endoscopic management modality for congenital membranous stenosis in the third portion of the duodenum. Standard approaches to duodenal stenosis in newborns include a laparotomy with an enteroenterostomy, bypassing the obstruction, or a duodenoduodenostomy with excision. We successfully developed a modification of the endoscopic treatment modality for congenital duodenal diaphragm.