Protein Complex of Fibrinogen Gamma Chain and Complement Factor H in Ovarian Cancer Patient Plasma.

BACKGROUND: In the plasma of an advanced cancer patient, fibrinogen is sometimes increased with possible effects on red blood cells (RBCs). MATERIALS AND METHODS: The plasma fraction deteriorating osmotic resistance of RBCs was separated from a patient's plasma with advanced ovarian cancer by phenyl-sepharose column chromatography and analyzed with gel filtration chromatography. RESULTS: In the plasma fraction, we found a protein reactive against whole fibrinogen with a molecular weight higher than that of intact fibrinogen from a healthy volunteer. The-high molecular weight protein was immunoractive to an antibody against fibrinogen gamma chain but not to an antibody against alpha or beta chain. Complement factor H, identified by N-terminal sequencing of a 150-kDa protein separated from the protein, was also eluted from anti-fibrinogen gamma immunoaffinity column. CONCLUSION: Fibrinogen gamma chain and complement factor H were found to be bound as a protein complex in the plasma of a patient with advanced ovarian cancer.

Human Papillomavirus E6 Knockdown Restores Adenovirus Mediated-estrogen Response Element Linked p53 Gene Transfer in HeLa Cells.

The p53 gene is inactivated by the human papillomavirus (HPV) E6 protein in the majority of cervical cancers. Treatment of HeLa S3 cells with siRNA for HPV E6 permitted adenovirus-mediated transduction of a p53 gene linked to an upstream estrogen response element (ERE). Our previous study in non-siRNA treated HHUA cells, which are derived from an endometrial cancer and express estrogen receptor ß, showed enhancing effects of an upstream ERE on adenovirus-mediated p53 gene transduction. In HeLa S3 cells treated with siRNA for HPV E6, adenovirus-mediated transduction was enhanced by an upstream ERE linked to a p53 gene carrying a proline variant at codon 72, but not for a p53 gene with arginine variant at codon 72. Expression levels of p53 mRNA and Coxsackie/adenovirus receptor (CAR) mRNA after adenovirus-mediated transfer of an ERE-linked p53 gene (proline variant at codon 72) were higher compared with those after non-ERE-linked p53 gene transfer in siRNA-treated HeLa S3 cells. Western blot analysis showed lower ß-tubulin levels and comparatively higher p53/ß-tubulin or CAR /ß-tubulin ratios in siRNA-treated HeLa S3 cells after adenovirus-mediated ERE-linked p53 gene (proline variant at codon 72) transfer compared with those in non-siRNA-treated cells. Apoptosis, as measured by annexin V binding, was higher after adenovirus-mediated ERE-linked p53 gene (proline variant at codon 72) transfer compared with that after non-ERE-linked p53 gene transfer in siRNA-treated cells.

Potential diagnostic biomarkers: differential expression of LMP2/ß1i and cyclin B1 in human uterine leiomyosarcoma.

AIMS AND BACKGROUND: Whilst most uterine smooth muscle neoplasms are benign, uterine leiomyosarcoma (Ut-LMS) is extremely malignant with a high incidence of metastasis and recurrence. Gynecological tumors are often associated with female hormone secretion, but no strong link has been detected between human Ut-LMS and the hormonal environment. In fact, the risk factors for Ut-LMS are poorly understood. In addition, no diagnostic biomarkers for differentiating between leiomyoma, a benign tumor, and malignant Ut-LMS have been found. Interestingly, mice that were homozygously deficient for LMP2/ß1i were found to spontaneously develop Ut-LMS and exhibited a Ut-LMS prevalence of ~40% by 14 months of age. Thus, analyzing potential risk factors for Ut-LMS (such as LMP2/ß1i) might aid the development of diagnostic biomarkers and clinical treatments for the condition. METHODS AND STUDY DESIGN: Fifty-seven patients (age range: 32-83 years) who had been diagnosed with uterine mesenchymal tumors were chosen from a pathological archive. Tissue samples from these patients were fixed in 10% buffered formalin, incubated in 4% paraformaldehyde for 8 hours, and embedded in paraffin. Tissue sections were stained with hematoxylin and eosin for standard histological examination or were subjected to further processing for immunohistochemical (IHC) examination. Serial Ut-LMS, bizarre leiomyoma, leiomyoma, and myometrium sections were subjected to IHC staining of ß-smooth muscle actin, estrogen receptor, cyclin B1, LMP2/ß1i, calponin h1, ki-67, tumor protein p53, and progesterone receptor. RESULTS: The Ut-LMS samples were positive for cyclin B1 and negative for LMP2/ß1i, while the opposite result was obtained for bizarre leiomyoma, leiomyoma, and myometrium samples. CONCLUSIONS: The expression pattern of LMP2/ß1i and cyclin B1 might be a diagnostic biomarker for human Ut-LMS. Studies of the biological roles of LMP2/ß1i and/or cyclin B1 could lead to the elucidation of new targets for therapies against Ut-LMS.

Pregnancy complicated by cervical varix and low-lying placenta: a case report.

We present a case of cervical varix and low-lying placenta. A cesarean section was performed because of the risk of bleeding with vaginal delivery; hemostasis was achieved using z sutures at the bleeding points. After delivery, the cervical varix decreased dramatically in size. It is important to recognize the clinical features and available treatments for cervical varix.

Pathological findings in a case of failed uterine artery embolization for placenta previa.

The reported success rate of uterine artery embolization (UAE) for obstetrical hemorrhage is more than 90%. We experienced a case of failed UAE for postpartum hemorrhage, although an embolic particle was found pathologically in the uterine vessels without coagulation. A 42-year-old woman (gravida 7, para 2) with placenta previa had genital bleeding at 35 weeks of gestation, and cesarean section was performed. We immediately added UAE aiming to reduce massive bleeding after the cesarean section, successful embolization of the bilateral uterine arteries and internal iliac arteries were confirmed by angiography regardless the vital sign was recovered with an appropriate amount of transfusion; the massive bleeding recurred after 1 hour of UAE. Hysterectomy was performed and pathological findings of the uterus showed that there was no coagulation in the vessels, which was supposed to be observed by the effect of gelatin sponge. In addition, despite the fact that no coagulation was found, only one gelatin sponge was found in 16 slices of the uterine wall specimens. We speculate that thrombotic materials were caught in vasoconstricted vessels triggered by hypovolemic shock due to acute blood loss, and then the gelatin sponge could be washed out after recovering to normalized circulation status leading to recurrent massive hemorrhage.

Presentation of a Patient who Underwent Fertility-Sparing Surgeries for Contralateral Recurrence of Ovarian Immature Teratoma with Gliomatosis Peritonei.

We report a patient who has maintained a regular menstrual cycle despite undergoing cystectomy and chemotherapy for contralateral recurrence of ovarian immature teratoma with gliomatosis peritonei. We initially performed a fertility-sparing right salpingo-oophorectomy, omentectomy and peritoneal biopsy for immature teratoma with gliomatosis peritonei, with adjuvant chemotherapy; we performed a left ovarian cystectomy and peritoneal biopsy for mature cystic teratoma with gliomatosis peritonei 16 months after the first surgery, a fertility-sparing left ovarian cystectomy and peritoneal biopsy for contralateral recurrence of ovarian immature teratoma with gliomatosis peritonei 60 months after the first surgery, and a left ovarian cystectomy and peritoneal and external iliac lymph node biopsy for endometrial cyst with gliomatosis peritonei 71 months after first surgery. The peritoneal gliomatosis lesions gradually decreased through the 4 surgeries over 8 years. The patient has maintained a regular menstrual cycle and currently shows no evidence of disease.

Association of copper transporter expression with platinum resistance in epithelial ovarian cancer.

BACKGROUND: Copper transporters (CTR) also regulate the cellular transport of platinum drugs, but their role in platinum resistance of ovarian cancer has not been elucidated. MATERIALS AND METHODS: CTR expression in ovarian cancer tissues resected from patients treated by platinum-based chemotherapy was evaluated immunohistochemically. CTR2 expression in ovarian cancer cells was inhibited by bathocuproine disulfonate, and the changes in cisplatin sensitivity were examined. RESULTS: CTR2 expression was increased in chemoresistant patients, but not significantly. However, the CTR2/CTR1 ratio was significantly increased in chemoresistant patients. Cases with positive CTR2 expression or positive CTR2/CTR1 ratio had poor prognoses. When the CTR2 expression in ovarian cancer cells was suppressed, sensitivity to cisplatin was significantly increased. CONCLUSION: These data suggest that CTR2 contributes to platinum resistance in ovarian cancer. The CTR2/CTR1 ratio is a useful marker for platinum sensitivity and a potential prognostic factor in patients with ovarian cancer.

Clinical management of atypical polypoid adenomyoma of the uterus. A clinicopathological review of 29 cases.

OBJECTIVE: The clinical management of atypical polypoid adenomyoma (APAM) of the uterus remains to be established. We collected APAM cases, reviewed the clinicopathological features, and discussed the clinical management. METHODS: Twenty-nine patients with APAM were identified by searching the tumor registry of the Japan Clinical Oncology Group (JCOG). Clinical information and histological specimens were obtained from 13 institutional members of the JCOG, and a central pathological review was performed. RESULTS: The mean age of the patients was 38 years (range, 22-58). Squamous metaplasia was present in 19 cases (65.5%), and well-differentiated endometrioid adenocarcinoma coexisted in 5 cases (17.2%). Primary treatment consisted of dilatation and curettage in 9 patients (31.0%), vaginal resection in 2 patients (6.9%), hysteroscopic transcervical resection (TCR) using hysteroscopy in 10 patients (34.5%), and hysterectomy in 8 patients (27.6%). There were recurrences in 5 (23.8%) of the 21 cases in which fertility was preserved, and the recurrent rate was 10% (1/10) in patients those were treated with TCR and 36.4% (4/11) in those the other treatment options were selected. All patients were alive after primary treatment (a mean follow-up period was 39.6 months; range, 1-202). CONCLUSION: The clinical outcome of APAM is benign. However, differential diagnosis should be performed because of its histological similarity to invasive endometrial carcinoma and the possibility of coexistence with other endometrial neoplasms. TCR is a recommended diagnostic and treatment option for patients who desire to preserve fertility.

Pegylated liposomal doxorubicin for platinum-resistant or refractory Müllerian carcinoma (epithelial ovarian carcinoma, primary carcinoma of Fallopian tube and peritoneal carcinoma): A single-institutional experience.

The aim of the present study was to evaluate the efficacy and safety of pegylated liposomal doxorubicin (PLD) in patients with Müllerian carcinoma treated at our hospital. Nineteen patients with platinum-resistant Müllerian carcinoma were treated with intravenous PLD 50 mg/m(2) every 4 weeks. Tumor response was assessed by MRI following every 2-3 cycles of treatment. The severity of adverse events was assessed according to the Common Terminology Criteria for Adverse Events (v3.0). The best overall responses in the 19 patients were identified as 5 partial responses (PR), 6 stable diseases (SD) and 8 progressive diseases (PD). Response rate was 26.3%. The proportion of patients with CR, PR or SD was 57.9%. The median time to progression was 188.0 days. The median survival time was 381.0 days. Toxicity grades were identified as one grade III hand-foot syndrome, two grade III neutropenia, one grade IV hand-foot syndrome, one grade IV stomatitis and one grade IV neutropenia. The present study confirmed that PLD is an effective drug when administered as a salvage therapy for the treatment of Müllerian carcinoma and is associated with a reduced toxicity profile compared with current therapeutic options.

Outcomes of traditional prolapse surgery for pelvic organ prolapse repair at a single center.

OBJECTIVE: To evaluate the clinical outcomes of traditional surgical approaches to pelvic organ prolapse. METHODS: From January 20, 2000, to March 24, 2009, 182 patients underwent traditional prolapse surgery at Osaka City University Graduate School of Medicine, Osaka, Japan, after prolapse assessment by the pelvic organ prolapse quantification system. Incontinence, defined as more than 2 g of urine leaked in a 1-hour pad test; posterior urethrovesical angle, using chain cystography; and quality of life (QOL), using a face visual analog scale, were assessed 1, 3, 6, and 12 months postoperatively, and then annually for 2 more years, in the 167 patients who completed follow-up. The presence of lower urinary tract symptoms was determined preoperatively and postoperatively. RESULTS: The anatomic and subjective recurrence rates were 21.0% and 6.0%. There was a significant difference in time to recurrence between anatomic and subjective prolapse. The posterior urethrovesical angle was improved postoperatively. The prevalence of lower urinary tract symptoms was 67.1% preoperatively and decreased to 28.2%, 17.2%, 16.2%, 15.6%, and 15.6% at 1 month, 6 months, 1 year, 2 years, and 3 years, respectively, and QOL was improved postoperatively regardless of anatomic recurrence. CONCLUSION: Anatomic support and QOL were satisfactory after traditional prolapse surgery.

Expression of mitotic-arrest deficiency 2 predicts the efficacy of neoadjuvant chemotherapy for locally advanced uterine cervical cancer.

We previously reported satisfactory therapeutic results when using cisplatin-based cyclic balloon-occluded arterial infusion chemotherapy as neoadjuvant chemotherapy (NAC), which enabled hysterectomy to be performed for patients with locally advanced cervical cancer. Mitotic arrest deficiency 2 (MAD2) is a key component of the mitotic spindle checkpoint pathway. The expression of MAD2 is associated with tumor progression and resistance to chemotherapy. Therefore, the aim of the present study was to examine whether the expression of MAD2 is related to the efficacy of NAC for locally advanced uterine cervical cancer. We reviewed 53 cases of locally advanced uterine cervical cancer (stage IIIa-IIIb; based on the International Federation of Gynecology and Obstetrics criteria). These patients were initially treated at Osaka City University Medical School Hospital, Japan, from 1995 to 2008 and were under 70 years old. Tumor samples were obtained by biopsy prior to NAC. Cases were divided into two groups: one group in which NAC was effective, surgery was possible and radiotherapy was performed (NAC+OP+R group; n=33), and another group in which NAC was ineffective and radiation therapy was performed (NAC+R group; n=20). MAD2 expression was examined in paraffin-embedded sections using the avidin-biotin peroxidase complex method. The results showed that MAD2 expression was significantly higher in the NAC+R group compared to the NAC+OP+R group (P<0.001). There was no significant difference in overall survival between the two groups, although the prognosis for the NAC+OP+R group tended to be slightly better (P=0.064). Taken together, these results suggest that the expression of MAD2 may predict the efficacy of NAC as a treatment for locally advanced uterine cervical cancer.

Prediction of the shrinking rate of uterine leiomyoma nodules using needle biopsy specimens.

OBJECTIVE: To determine correlations between shrinkage of uterine leiomyomas after treatment with GnRH agonists (GnRH-a) or menopause and expression levels of estrogen receptors (ER), progesterone receptors (PR), and vascular endothelial growth factor (VEGF). DESIGN: Cohort study. SETTING: University teaching hospital. PATIENT(S): A total of 26 women with uterine leiomyoma. INTERVENTION(S): Ten women were treated with buserelin acetate injection (1.8 mg), four courses every 4 weeks, and 16 women went into menopause naturally. MAIN OUTCOME MEASURE(S): Tumor shrinkage rates determined from magnetic resonance images taken before and after GnRH-a therapy and before and after natural menopause; immunohistochemical analysis of ER, PR, and VEGF in uterine leiomyoma biopsy specimens taken before intervention or within 6 months before menopause. RESULT(S): Shrinkage rates of uterine leiomyomas were positively correlated with expression levels of ER in women treated with GnRH-a and in postmenopausal women managed conservatively, and with VEGF expression in women treated with GnRH-a. There were no significant correlations with PR expression levels in either group. CONCLUSION(S): Estrogen plays the predominant role in myoma shrinkage for women with GnRH-a therapy and naturally menopausal women.

Expression of the mitotic-arrest deficiency 2 is associated with chemotherapy resistance in ovarian serous adenocarcinoma.

Mitotic-arrest deficiency 2 (MAD2) is a key component of spindle assembly checkpoint (SAC) function; SAC mediates spindle microtubule attachment to kinetochores on chromosomes and chromosomal segregation during mitosis. To determine whether MAD2 expression is associated with chemotherapy resistance in ovarian serous adenocarcinoma, we reviewed tumor samples from 41 cases of ovarian serous adenocarcinoma at Osaka City University Medical School Hospital (Osaka, Japan), 2000-2007. Of the 41 cases, 24 were recurrent and 17 were not recurrent. Expression of MAD2 was investigated in paraffin-embedded sections using a MAD2 antibody. Quantitative analysis of MAD2 expression gave mean weighted scores of 4.3 for the relapsed group and 7.2 for the relapse-free group; the expression was, thus, significantly greater in the relapse-free group compared to the relapsed group (P=0.023). When the 41 cases were classified into low- and high-expression, these classifications showed no significant difference in terms of progression-free survival (P=0.0685), however, overall survival for the low-expression group was significantly shorter than that of the high-expression group (P=0.0188). The present study implies that MAD2 expression levels can indicate sensitivity to anticancer agents, and risk for recurrence.

Potential role of LMP2 as an anti-oncogenic factor in human uterine leiomyosarcoma: morphological significance of calponin h1.

Uterine leiomyosarcoma (LMS) is a highly metastatic smooth muscle neoplasm for which calponin h1 is suspected to have a biological role as a tumor-suppressor. We earlier reported that LMP2-null mice spontaneously develop uterine LMS through malignant transformation of the myometrium, thus implicating this protein as an anti-tumorigenic candidate as well. In the present study, we show that LMP2 may negatively regulate LMS independently of its role in the proteasome. Moreover, several lines of evidence indicate that although calponin h1 does not directly influence tumorigenesis, it clearly affects LMP2-induced cellular morphological changes. Modulation of LMP2 may lead to new therapeutic approaches in human uterine LMS.

Mitotic arrest deficiency 2 induces carcinogenesis in mucinous ovarian tumors.

Mitotic arrest deficiency 2 (MAD2) is a key component of the mitotic spindle checkpoint pathway. A compromised mitotic spindle checkpoint results in an abnormal number of chromosomes. This is referred to as chromosomal instability, and has been reported in most types of human cancer. The aim of this study was to examine the expression of MAD2 in mucinous ovarian tumors exhibiting varying degrees of malignancy. We reviewed 128 cases of mucinous ovarian tumors initially treated at Osaka City University Medical School Hospital, Japan. Tumor samples were obtained following surgery. The cases were divided into three groups: benign (group B; n=30), borderline malignant (group BM; n=55) and malignant (group M; n=43). MAD2 expression was examined in paraffin-embedded sections using the avidin-biotin peroxidase complex method. Results showed MAD2 expression to be significantly greater in group M compared to groups B and BM (P<0.05). In addition, there was a moderate correlation between MAD2 expression and the degree of malignancy (r=0.51, P<0.05). However, when the samples in group M were classified according to a low or high expression of MAD2, no difference was observed in terms of overall survival. These findings suggest that the overexpression of MAD2 may be correlated to carcinogenesis in mucinous ovarian tumors.

Reference ranges for time-related analysis of ductus venosus flow velocity waveforms in singleton pregnancies.

OBJECTIVES: To investigate time intervals of ductus venosus (DV) flow velocity waveforms (FVW) in correlation to fetal heart rate and gestational age and to construct reference ranges for the second and third trimester. Furthermore, we investigate time intervals of FVW through the tricuspid valve. METHODS: Flow velocity waveforms of the DV and through the tricuspid valve were recorded in 135 normal singleton fetuses between 17 and 38 weeks' gestation. Time intervals for systolic (S) and early diastolic (D) peaks were analyzed regarding acceleration time (acc-S for S, acc-D for D) and deceleration time (dec-S for S, dec-D for D), respectively. Similarly, time intervals for both peaks of right ventricular inflow were analyzed regarding acceleration time (acc-E for E-wave, acc-A for A-wave) and deceleration time (dec-E for E-wave, dec-A for A-wave), respectively. RESULTS: In the DV, acc-D and dec-D increased significantly with gestational age. In tricuspid valve, acc-E and acc-A showed a significant increase with gestational age. All parameters except acc-S showed significant negative correlations with fetal heart rate. CONCLUSION: With advancing gestational age, prolongation of the diastolic phase of DV-FVW and of the E-wave of tricuspid flow was observed, suggesting maturation of ventricular diastolic function. Time-related analysis of Doppler signals of DV-FVW may provide detailed insights into fetal cardiac function.

Maternal dietary patterns in pregnancy and fetal growth in Japan: the Osaka Maternal and Child Health Study.

Maternal nutritional status during pregnancy is an important determinant of fetal growth. Although the effects of several nutrients and foods have been well examined, little is known about the relationship of overall maternal diet in pregnancy to fetal growth, particularly in non-Western populations. We prospectively examined the relationship of maternal dietary patterns in pregnancy to neonatal anthropometric measurements at birth and risk of small-for-gestational-age (SGA) birth among 803 Japanese women with live-born, singleton, term deliveries. Maternal diet in pregnancy was assessed using a validated, self-administered diet history questionnaire. Dietary patterns from thirty-three predefined food groups (g/4184 kJ) were extracted by cluster analysis. The following three dietary patterns were identified: the 'meat and eggs' (n 326), 'wheat products', with a relatively high intake of bread, confectioneries and soft drinks (n 303), and 'rice, fish and vegetables' (n 174) patterns. After adjustment for potential confounders, women in the 'wheat products' pattern had infants with the significantly lowest birth weight (P = 0·045) and head circumference (P = 0·036) among those in the three dietary patterns. Compared with women in the 'rice, fish and vegetables' pattern, women in the 'wheat products' pattern had higher odds of having a SGA infant for weight (multivariate OR 5·2, 95 % CI 1·1, 24·4), but this was not the case for birth length or head circumference. These results suggest that a diet high in bread, confectioneries, and soft drinks and low in fish and vegetables during pregnancy might be associated with a small birth weight and an increased risk of having a SGA infant.

An upstream estrogen response element linked to exogenous p53 tumor suppressor gene expression differentiates effects of the codon 72 polymorphism.

The objective of this study was to assess the effects of an upstream estrogen response element (ERE) on exogenous p53 tumor suppressor gene with a codon 72 polymorphism about which there have been controversial reports in relation to cancer risk. The p53 gene (bases 166-1143 from start codon) with the codon 72 polymorphism, inserted into the pIRES-hrGFP II plasmid with or without upstream ERE, were transfected into HHUA endometrial cancer cells expressing the estrogen receptor. The ERE-linked p53 gene with the proline variant at codon 72 showed lower transfection rates than the gene without ERE or with the arginine variant at codon 72. p21 expression was significantly higher in HHUA cells transfected with the proline variant gene than in those transfected with the arginine variant gene. We consider that the presence of an upstream ERE promotes the transcriptional effects of the exogenous p53 gene with the proline variant, which strengthens the expression of p21, and results in lower transfection rates through cell cycle inhibition.

Claudin-4: a potential therapeutic target in chemotherapy-resistant ovarian cancer.

BACKGROUND: Claudin-4, a component of the tight junction, plays an important role in tumorigenesis and metastasis of ovarian cancer, but its role in platinum resistance has not been elucidated. MATERIALS AND METHODS: Claudin-4 expression in ovarian cancer cells was inhibited and the changes in cisplatin sensitivity were examined. Fluorescence-labeled cisplatin was used to examine whether inhibition of claudin-4 changed the cellular accumulation of cisplatin. Claudin-4 expression in ovarian cancer tissue resected from the patients surgically was evaluated immunohistochemically. RESULTS: Suppression of claudin-4 resulted in a significant increase of cisplatin sensitivity and cellular accumulation of fluorescence-labeled cisplatin. Claudin-4 expression was significantly greater in ovarian cancer tissue from chemoresistant patients compared to chemosensitive patients. The overall survival was significantly shorter for claudin-4-positive than claudin-4-negative cases. CONCLUSION: These data suggest that claudin-4 contributes to platinum resistance in ovarian cancer and may be a potential target in the treatment of platinum-resistant tumors.