Biweekly administration of docetaxel and carboplatin for advanced or recurrent endometrial and ovarian carcinomas.

OBJECTIVE: To examine efficacy and safety of biweekly administration of docetaxel and carboplatin for advanced or recurrent en- dometrial and ovarian carcinomas. MATERIAL AND METHODS: The recommended doses were determined in the phase I study. In the phase II feasibility study, the primary end-point was safety, and the secondary end-point was response rate and progression-free survival (PFS). RESULTS: The recommended doses of docetaxel and carboplatin were determined to be 45 mg/n(2) and AUC 3.0, respectively, in phase I study. In phase II feasibility study, no treatment-related death was observed. Most non-hematotoxicity cases were mild or moderate. Grade 4 neutropenia was confirmed in 13 patients (31.0%), whereas all cases showed tolerability with 2.6 days delay of anticancer drugs administration in both groups. Response rate was 55.0% in the ovarian carcinoma group, and average PFS was 8.7 months. In the endometrial carcinoma group, response rate was 50.0% and average PFS was 32.0 months. CONCLUSION: The present results showed that biweekly administration of docetaxel and carboplatin for advanced and recurrent endometrial and ovarian carcinomas results in acceptable side effects, response rate, and PFS.

Evaluation of the human papillomavirus mRNA test for the detection of cervical lesions in Japan.

AIMS: For the screening of cervical abnormalities, human papillomavirus (HPV) DNA testing is widely used along with Papanicolaou (Pap) testing. Although the sensitivity of the HPV DNA testing is good, its specificity is relatively low. In the present study, the authors evaluated the use of the Gen-Probe APTIMA HPV Assay for the detection of HPV mRNA and compared it with HPV DNA testing. MATERIALS AND METHODS: Liquid cervical Pap specimens collected from 410 women were assessed using the APTIMA test, the Qiagen Hybrid Capture 2 HPV DNA (HC2) Test, and the AMPLICOR HPV Test. RESULTS: The sensitivity and specificity for the detection of high-risk HPV were 85.6% and 99.2% for the APTIMA test, 94.1% and 98.4% for the HC2 test, and 90.2% and 95.7% for the AMPLICOR test, respectively. As the severity of the cervical lesion progressed, the positive rate of the three tests indicated a similar increase. The clinical sensitivity and specificity for the detection of squamous intraepithelial lesion (SIL) were 91.2% and 84.2% for the APTIMA test, 94.5% and 80.4% for the HC2 test, and 87.9% and 78.2% for the AMPLICOR test, respectively. CONCLUSION: The APTIMA is sensitive and specific for the detection of high-risk HPV. In the specimens with SIL, the APTIMA test is more specific than the HC2 and the AMPLICOR tests. This indicates that the APTIMA test may improve patient management and reduce the cost of screening.

A case of squamous cell carcinoma arising from endometriosis of the ovary.

Ovarian endometriosis sometimes develops into ovarian cancer, especially clear cell adenocarcinoma and endometrioid adenocarcinoma. However, endometriosis rarely develops into squamous cell carcinoma. We present a case of squamous cell carcinoma arising from endometriosis. A 47-year-old Japanese woman was given a diagnosis of ovarian squamous cell carcinoma arising from endometriosis. She was treated with combination chemotherapy consisting of paclitaxel and carboplatin once every three weeks. Four months after the initial chemotherapy, multiple liver tumors appeared, and her treatment was changed to palliative therapy. Based on this case, in which ovarian squamous cell carcinoma arose from endometriosis, endometriosis should be followed-up strictly.

A case of uterine cervical carcinosarcoma recurrence who obtained a clinically complete response by ifosfamide, doxorubicin and cisplatin.

Cervical carcinosarcoma (CS) is a rare gynecologic tumor. The histogenesis, clinical features, and optimal treatment remain unclear. We report a case of cervical CS recurrence to the right lung, which had complete response by treating with ifosfamide, doxorubicin and cisplatin (IAP). A 61-year-old woman underwent semi-radical hysterectomy, bilateral salpingo-oophorectomy, and pelvic lymphadenectomy for CS of the uterine cervix. Eleven months later, the patient presented with left pulmonary metastasis. She refused debulking surgery and had chemotherapy with IAP. After four cycles of chemotherapy, the metastatic tumor completely disappeared. Unfortunately, a re-recurrent tumor was seen in the same lung area six months after IAP. Eventually, she died 39 months after surgery.

Microbial community of anammox bacteria immobilized in polyethylene glycol gel carrier.

Anaerobic ammonium oxidation (anammox) is a recently discovered microbial pathway in the biological nitrogen cycle and a new cost-effective way to remove ammonium from wastewater. We have so far developed new immobilization technique that anammox bacteria entrapped in polyethylene glycol (PEG) gel carrier. However, fate and behavior of anammox bacteria in a gel carrier is not well understood. In the present study, we focused on the population changes of anammox bacteria in a gel carrier. Three specific primer sets were designed for real-time PCR. For quantification of anammox bacteria in a gel carrier, real-time PCR was performed. The anammox bacteria related to HPT-WU-N03 clone were increased the rate in anammox population, and found to be a major population of anammox bacteria in a gel carrier. Furthermore, from the results of nitrogen removal performance and quantification of anammox bacteria, the correlation coefficient between copy numbers of anammox bacteria and nitrogen conversion rate was calculated as 0.947 in total anammox population. This is the first report that population changes of anammox bacteria immobilized in a gel carrier were evaluated.

Early growth response-1 mediates up-regulation of telomerase in placenta.

Telomerase is thought to play a very important role in oncogenesis. It is also believed to wind back the "mitotic clock" which leads to ageing and enable permanent cell division. We evaluated telomerase activity in chorionic tissues, with particular attention to the early growth response-1 (EGR-1) gene, the importance of what was recently shown by Khachigian et al. We started our study by evaluating the relationship between activation of transcription of the human telomerase reverse transcriptase (hTERT) gene and EGR-1 gene. For this purpose, we first evaluated telomerase activity using the villous cancer cell lines JAR and JEG-3. We then demonstrated that EGR-1 plays an important role in activation of the transcription of hTERT by luciferase assay using hTERT promoter constructs. As a result of further computer analysis, we discovered a site postulated to be an EGR-1 consensus binding site at -273 to -281 in the hTERT promoter region. With forced expression of EGR-1, an increase in hTERT protein concentration was detected on Western blot analysis, while marked high expression of hTERT mRNA was observed by reverse transcriptase polymerase chain reaction. Furthermore, we evaluated the expression of EGR-1 and hTERT at the mRNA level in the placenta during the first, second and third trimesters of pregnancy and in patients with preeclampsia. Expression of EGR-1 and hTERT in the chorion increased in the first trimester of pregnancy and decreased later. Increased expression was noted in the placenta of patients with preeclampsia. The present findings suggest that EGR-1 plays an important role in activating the transcription of hTERT, showing that activation of the transcription of hTERT by EGR-1 is involved in the trophoblast growth mechanism.

Synthesis of placental protein 12 by decidua from early pregnancy.

The synthesis and secretion of placental protein 12 (PP12) by early pregnancy decidua and trophoblast were studied in vitro from tissues obtained by curettage during elective termination of pregnancy (weeks 8-14). The tissue explants were incubated in Ham's F-10 medium for a 27-h period, and the PP12 levels in media and tissue homogenates were measured by RIA. De novo synthesis of PP12 was assessed by measuring the incorporation of radioactivity into PP12 after 20 h of incubation of tissues with 20 microCi/ml [35S]methionine. PP12 from the culture medium was immunoprecipitated with anti-PP12(A) antiserum, and the immunoprecipitate was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The initial tissue content of PP12 was 10- to 72-fold higher in decidua than in trophoblast. The total amount of radioimmunoassayable PP12 released into medium by decidual explants during the 27-h incubation period together with that present in the tissues at the end of incubation exceeded the initial tissue content by 242.7 +/- 63.7% (mean +/- SE). Only small amounts of PP12 were detected in media from trophoblast cultures. During the first 7 h of incubation, inclusion of cycloheximide had no effect on PP12 release by decidual explants in three of four experiments. Between 7 and 27 h, the amount of PP12 released by cycloheximide-treated tissues was 20.0 +/- 7% of that released by control tissues (P less than 0.01). Cycloheximide had no effect on PP12 release by trophoblasts. Decidual explants incorporated [35S]methionine into PP12, but trophoblasts did not. In sodium dodecyl sulfate-gel electrophoresis, the newly synthesized PP12 comigrated with the major band of purified PP12 corresponding to mol wt 29,000. These data clearly confirm that PP12 is a protein of decidual rather than trophoblastic origin, and indicate that decidua from early pregnancy has the ability to synthesize it.

The correlation of TERT expression with c-myc expression in cervical cancer.

Recently putative catalytic telomerase subunit was identified as telomerase reverse transcriptase (TERT). Several reports showed that TERT expression correlated with telomerase activity. It has also been found that c-myc can induce telomerase activation through TERT expression. We examined expression of TERT and c-myc and their correlation in cervical cancers by reverse transcription-polymerase chain reaction (RT-PCR). It was found that TERT and c-myc expression was observed mostly in malignancies and expression of c-myc was concordant for positivity and negativity with TERT. These results support recent studies that c-myc expression is closely associated with TERT and telomerase activity. c-myc up-regulation may play an important role in activation of TERT and telomerase.

Expression of telomerase subunits and localization of telomerase activation in hydatidiform mole.

Activation of telomerase compensating for the loss of telomeres has been implicated in human cell immortalization and carcinogenesis. Telomeric repeat amplification protocol (TRAP) assay can be used to detect telomerase activity in a variety of malignant tumours, including those of the female reproductive tract which have been found to have high levels of telomerase activity. However, it is unclear whether all the cells or only a subset of cells within a tumour have telomerase activity. To determine the regulation mechanism of telomerase activity in hydatidiform moles, we studied telomerase activity at the single cell level (using an in situ TRAP assay), and expression of TLP1 (telomerase protein 1), TERC (telomerase RNA component) and TERT (telomerase reverse transcriptase component). Expression of TERC and TLP1 was observed in all normal chorionic villi, as well as in trophoblastic diseases, and various cell lines irrespective of telomerase activity. TERT expression was observed in trophoblastic diseases and normal chorionic villi with telomerase activity but not in normal chorionic villi without telomerase activity, except in some cases in the present series, indicating that TERT expression is closely associated with telomerase activity. Upregulation of TERT expression may thus play an important role in telomerase reactivation.

Expression and activity of matrix metalloproteinase 2 and 9 in human trophoblasts.

Indiscriminate invasion upon the endometrium by normal trophoblasts is strictly regulated unlike that by choriocarcinoma cells. In this study, we focused on the activity of matrix metalloproteinase (MMP)-2 and MMP-9 as parameters of invasion in normal human placenta. In situ hybridization (ISH), immunohistochemical staining (IH) and film in situ zymography (FIZ) were performed to identify cells having MMP-2 or MMP-9 expression and activity. Purified cytotrophoblasts (CTs) were used to examine the expression and activity of MMP-2 and MMP-9, and their invasive ability. In first trimester placental tissue, the MMP-2 expression was observed in extravillous trophoblasts (EVTs), and MMP-9 mainly in villous cytotrophoblasts (VCTs). FIZ revealed marked gelatinase activity in the EVTs which MMP-2 expression was observed in. In full-term placental tissue, the MMP-2 expressions was observed in the EVTs similar to that in first trimester, whereas the gelatinase activity in these cells was decreased or completely lost. Using purified CTs, the gelatinase activity was marked in early CTs, but not term CTs. Invasive ability of early CTs was inhibited by tissue inhibitor of metalloproteinase (TIMP)-2 and MMP-2 antibody in a dose dependent manner. These suggests that the invasive ability of trophoblasts may be regulated by the enzyme activity of gelatinases, especially MMP-2.

Comparison of telomerase activity in normal chorionic villi to trophoblastic diseases.

Trophoblasts are derived from the normal placenta, and they infiltrate into the endometrium and the maternal blood vessels under strict control but, unlike malignant cells, never metastasize. To understand the proliferative characteristics of trophoblasts and its related disorders, we assessed telomerase activity in chorionic villi obtained from 27 normal individuals, 9 hydatidiform moles, and 2 choriocarcinomas. Telomerase activity was detected in 13/27 (48%) normal chorionic villi samples. The detectability and the level of telomerase activity depended on gestational age; 8/10 (80%) villi samples in the first trimester (relative telomerase activity; 1.77 +/- 1.37), whereas 2/8 (25%) villi samples in the second trimester (0.78 +/- 1.52) and 3/9 (33%) in the third trimester (0.28 +/- 0.43) had telomerase activity. Telomerase activity of normal chorionic villi in the first trimester was higher than that of the third trimester (P = 0.0251). In contrast, all mole samples had increased telomerase activity compared to normal villi (3.17 +/- 2.81, P = 0.0152). Thus, a relationship may exist among cell proliferation, telomerase activity, and progression to trophoblastic disease.

Mutation and transcription analyses of the p63 gene in cervical carcinoma.

Genetic mutation of p53, which monitors DNA damage and operates cellular checkpoints, is a major factor in the development of human malignancies. A novel gene p63/p73L/p51, encoding a protein with significant homology to p53 and p73, was recently identified at 3q27-9. To investigate the penetration of p63 in cervical carcinogenesis, mutation and transcription analyses of p63 were performed in cervical carcinoma. A certain isotype of p63 called TAp63gamma encodes the acidic N-terminus and possesses a short C-terminus. Using reverse transcriptase-polymerase chain reaction-single strand conformation polymorphism (RT-PCR-SSCP) analysis for TAp63gamma, one mutation was found in the cervical carcinoma cell line SKG-I. However, no mutations causing amino acid substitutions or frameshifts were found in 54 cases examined for TAp63gamma, which is thought to be a tumor suppressor gene. While cervical carcinomas tended to yield a positive signal in the RT-PCR reaction designed to amplify transcripts encoding the acidic N-terminus, normal cervix and cervical intraepithelial neoplasia (CIN) did not express this transcript. These data suggest that the p63 gene does not play an essential role as a tumor suppressor gene, but expression of TAp63gamma may be speculatively associated with tumor growth in cervical carcinogenesis.

[Establishment and characterization of the new cell line (EI) from a human endometrial adenocarcinoma].

A moderately-differentiated endometrial adenocarcinoma cell line(EI) was established from a surgical specimens obtained from a 55-year-old woman with endometrial carcinoma. This cell line could be transplanted to nude mice, where the cells showed the same histological type as the primary tumor. The doubling time of the cell line was 50.5 hours; the saturation density was 7.5 x 104 cells/cm2; the plating efficiency was 46%. This cell line was determined to produce TPA, but not other tumor markers, such as CA125 or CEA. Neither estrogen receptor, nor progesterone receptor was detected from the culture cell or the primary tumor. Chromosome analysis revealed that cells examined were all 46,XX, + 8,t(14q14q), and only cells with this karyotype were thought to be able to grow. From these results, it was suggested that a gene on No. 8 chromosome would be involved in the carcinogenesis of endometrial adenocarcinoma. Thus this cell line was thought to be useful for the clarification of gene conversion during the process of development of endometrial adenocarcinoma.

Regulation of decidualization in human endometrial stromal cells through exchange protein directly activated by cyclic AMP (Epac).

INTRODUCTION: Human endometrial stromal cells (ESCs) undergo differentiation during the decidualization process. Decidualization is characterized by their enhanced production of IGF binding protein-1 (IGFBP-1), prolactin (PRL), and the forkhead transcriptional factor FOXO1, and transformation into more rounded cells, during the secretory phase of the menstrual cycle and subsequent pregnancy. Protein kinase A (PKA)-mediated cAMP signaling is crucial for this process. The present study was undertaken to examine the involvement of a mediator of cAMP signaling, exchange protein directly activated by cAMP (Epac), in decidualization of cultured ESCs. RESULTS: Treatment of ESCs with the Epac-selective cAMP analog 8-CPT-2-OMe-cAMP (CPT) had no effect on IGFBP-1, PRL, and FOXO1 mRNA expression. However, CPT potentiated IGFBP-1 and PRL expression stimulated by the PKA-selective cAMP analog N(6)-Phe-cAMP (Phe) and activated Rap1, a downstream regulator of Epac signaling. Knock-down of Epac1, Epac2, or Rap1 significantly inhibited the Phe- or Phe/CPT-induced increase in IGFBP-1 and PRL expression, as well as Rap1 activation. Furthermore, CPT enhanced IGFBP-1 and PRL expression and the morphological differentiation induced by ovarian steroids, whereas Epac1, Epac2, or Rap1 knock-down suppressed these events. CONCLUSION: These data provide evidence for the involvement of the Epac/Rap1 signaling pathway in cAMP-mediated decidualization of human ESCs.

Effects of tissue conductivity and electrode area on internal electric fields in a numerical human model for ELF contact current exposures.

Contact currents flow through the human body when a conducting object with different potential is touched. There are limited reports on numerical dosimetry for contact current exposure compared with electromagnetic field exposures. In this study, using an anatomical human adult male model, we performed numerical calculation of internal electric fields resulting from 60 Hz contact current flowing from the left hand to the left foot as a basis case. Next, we performed a variety of similar calculations with varying tissue conductivity and contact area, and compared the results with the basis case. We found that very low conductivity of skin and a small electrode size enhanced the internal fields in the muscle, subcutaneous fat and skin close to the contact region. The 99th percentile value of the fields in a particular tissue type did not reliably account for these fields near the electrode. In the arm and leg, the internal fields for the muscle anisotropy were identical to those in the isotropy case using a conductivity value longitudinal to the muscle fibre. Furthermore, the internal fields in the tissues abreast of the joints such as the wrist and the elbow, including low conductivity tissues, as well as the electrode contact region, exceeded the ICNIRP basic restriction for the general public with contact current as the reference level value.