Phase 2 study of anastrozole in rare cohorts of patients with estrogen receptor/progesterone receptor positive leiomyosarcomas and carcinosarcomas of the uterine corpus: The PARAGON trial (ANZGOG 0903).

BACKGROUND: Aromatase inhibitors have been used empirically to treat a subset of patients with hormone receptor positive uterine leiomyosarcomas(LMS) and carcinosarcomas (UCS) mainly supported by retrospective data. We evaluated the activity of anastrozole in two rare cohorts; patients with recurrent/metastatic LMS and UCS enrolled in PARAGON, a basket trial of anastrozole in estrogen receptor (ER+)/progesterone receptor positive (PR+) gynecological cancers. METHOD: An investigator-initiated, single-arm, prospective open-label trial of anastrozole 1 mg/day in patients with ER &/or PR + ve LMS or UCS with measurable disease, treated until progression or unacceptable toxicity. Primary endpoint was clinical benefit (complete/partial response + stable disease) rate (CBR) at 3 months. Secondary endpoints include progression-free survival (PFS), quality of life and toxicity. RESULTS: 39 eligible patients were enrolled, 32 with LMS and 7 with UCS. For the LMS cohort CBR at 3 months was 35% (95% CI: 21-53%) with a median duration of clinical benefit of 5.8 months. Best response was a partial response in one patient. Two patients remained on treatment for more than one year. The median progression-free survival was 2.8 months (95% CI: 2.6-4.9). For the UCS cohort CBR at 3 months was 43% (95% CI: 16-75%) with a median duration of clinical benefit of 5.6 months. Stable disease was seen in 3 patients but no objective responses were seen. The median progression-free survival was 2.7 months (95% CI, 1.1-8.2). Safety was acceptable with 5/39 evaluable patients showing grade 3 toxicities. CONCLUSION: Whilst objective response rates with anastrozole are low, the clinical benefit rate and good tolerance suggests that aromatase inhibitor therapy may have a role in a subset of patients with metastatic LMS and UCS.

Lack of high-dose radiation mediated prostate cancer promotion and low-dose radiation adaptive response in the TRAMP mouse model.

Cancer of the prostate is a highly prevalent disease with a heterogeneous aetiology and prognosis. Current understanding of the biological mechanisms underlying the responses of prostate tissue to ionizing radiation exposure, including cancer induction, is surprisingly limited for both high- and low-dose exposures. As population exposure to radiation increases, largely through medical imaging, a better understanding of the response of the prostate to radiation exposure is required. Low-dose radiation-induced adaptive responses for increased cancer latency and decreased cancer frequency have been demonstrated in mouse models, largely for hematological cancers. This study examines the effects of high- and low-dose whole-body radiation exposure on prostate cancer development using an autochthonous mouse model of prostate cancer: TRansgenic Adenocarcinoma of the Mouse Prostate (TRAMP). TRAMP mice were exposed to single acute high (2 Gy), low (50 mGy) and repeated low (5 × 50 mGy) doses of X rays to evaluate both the potential prostate cancer promoting effects of high-dose radiation and low-dose adaptive response phenomena in this prostate cancer model. Prostate weights and histopathology were examined to evaluate gross changes in cancer development and, in mice exposed to a single 2 Gy dose, time to palpable tumor was examined. Proliferation (Ki-67), apoptosis, DNA damage (γ-H2AX) and transgene expression (large T-antigen) were examined within TRAMP prostate sections. Neither high- nor low-dose radiation-induced effects on prostate cancer progression were observed for any of the endpoints studied. Lack of observable effects of high- or low-dose radiation exposure suggests that modulation of tumorigenesis in the TRAMP model is largely resistant to such exposures. However, further study is required to better assess the effects of radiation exposure using alternative prostate cancer models that incorporate normal prostate and in those that are not driven by SV40 large T antigen.

False-positive TUNEL staining observed in SV40 based transgenic murine prostate cancer models.

The TRAMP (Transgenic Adenocarcinoma of the Mouse Prostate) and LADY (Probasin-large T antigen transgenic mouse) mice are widely used autochthonous models of prostate cancer. Both models utilise probasin promoters to direct androgen-regulated expression of oncogenic SV40 specifically to epithelial cells of the mouse prostate. The oncogenic processes and phenotypes which result mimic many features of human prostate cancer, making these transgenic mouse models useful experimental systems. The terminal deoxynucleotidyl transferase (Tdt)-mediated dUTP in situ nick end labelling (TUNEL) assay is a commonly used method for the detection of cells undergoing apoptosis. In this study, we demonstrate false-positive TUNEL staining in frozen prostate tissue from TRAMP and LADY mice, which was not observed in non-transgenic control animals and is not due to non-specific binding of labelled-dUTP substrate. The false-positive signal co-localised with large SV40 T-antigen expression. False-positive signal was apparent using multiple commercial TUNEL kits with different detection systems. These results caution against the use of the TUNEL assay for detection of apoptosis in frozen prostate tissue of large T-antigen based autochthonous transgenic models of prostate cancer.

Evaluation of external operculum loop tags to individually identify cage-cultured Atlantic halibut Hippoglossus hippoglossus in commercial research trials.

The growth, survival and tag retention of double-tagged [external FT4 lock-on (FT4) and internal passive integrated transponder (PIT)-tagged] Atlantic halibut Hippoglossus hippoglossus were compared to internal PIT-tagged controls in a randomized trial. The objective was to assess the suitability of these tags for monitoring the performance of individual fish in longitudinal trials under commercial cage-culture conditions in the lower Bay of Fundy, New Brunswick, Canada. The FT4 tags were chosen due to their similarity to tags used by investigators to track H. hippoglossus in the wild. A subset of the population randomly received an external FT4 tag inserted through the operculum and were monitored over a 1105 day period. The specific growth rate of FT4-tagged fish was significantly reduced in the first sea summer with no significant difference observed for the remainder of the trial. The differential growth in the first sea summer created a relative size advantage, permitting controls to increase in size significantly faster than FT4 fish in all subsequent periods. The FT4 tags did not significantly influence survival under normal commercial cage-culture conditions. Results, however, suggest that the survival of FT4-tagged H. hippoglossus may be compromised during stressful handling events. Tag retention of FT4 tags was acceptable with 76% of tags remaining at the end of the 1105 day trial. FT4 tags proved to be an effective method to identify individual H. hippoglossus, with the caveat that they seriously bias productivity measures in commercial research trials.

Adrenomedullin interacts with VEGF in endometrial cancer and has varied modulation in tumours of different grades.

OBJECTIVE: Endometrial cancer, in developed countries, is the most common malignancy of the female genital tract. Surgery and radiotherapy are successful in many patients but systemic and recurrent diseases have no consistently effective treatments, and for high grade advanced disease the prognosis is poor. The study investigated characteristics of adrenomedullin in endometrial cancer to assist in identifying targets for developing treatments. METHODS: Endometrial samples of women with and without cancer, and the Ishikawa cell line were used to investigate adrenomedullin mRNA regulation, peptide expression, adrenomedullin secretion and effects of adrenomedullin on VEGF secretion. RESULTS: Expression of adrenomedullin mRNA was upregulated compared to that in healthy post-menopausal endometria. Adrenomedullin secretion was increased by cobalt chloride in this study. Secretion was reduced by the naturally-occurring compounds, (-)-epigallocatechin gallate (EGCG) and 3,4',5-trihydroxystilbene (resveratrol), which we have previously demonstrated to also suppress VEGF secretion in endometrial tumour tissue. We noted, for the first time, that adrenomedullin enhanced VEGF secretion from tumour cells. CONCLUSIONS: Increased adrenomedullin expression may result in amplifying both tumorigenic and angiogenic activities. A substantial impact on growth of tumours may result in vivo as a consequence of the synergism between adrenomedullin and VEGF. Adrenomedullin, which has altered cellular characteristics in tumour compared to healthy tissue, offers an understudied target with potential to modify endometrial cancer behaviour, complementing other treatments.

Successful reproductive outcome following treatment of advanced small cell carcinoma of the ovary.

► Ovarian small cell carcinoma usually occurs in adolescents or young adults. ► Long term survival of advanced stage disease is extremely rare. ► Fertility may be conserved and should be considered as part of management.

Workers' exposure to dust, endotoxin and ß-(1-3) glucan at four large-scale composting facilities.

OBJECTIVES: To characterise compost workers' exposure to dust, endotoxin and ß-(1-3) glucan during various operational practices and investigate whether dust concentrations are a useful indicator of endotoxin exposure in compost workers. METHODS: This study assessed inhalable dust fractions, bacterial endotoxin and ß-(1-3) glucan in 117 personal samples and 88 ambient samples from four large-scale composting facilities. RESULTS: Employees' exposures to inhalable dust, endotoxin and ß-(1-3) glucan exhibited a large range. Inhalable dust was found to be generally low (GM 0.99 mg/m(3), GSD 2.99 mg/m(3)). Analysis of the biological component of the dust showed that employees' exposures to endotoxin were elevated (GM 35.10 EU/m(3), GSD 9.97 EU/m(3)). Employees' exposure to ß-(1-3) glucan was low (GM 0.98 ng/m(3), GSD 13.39 ng/m(3)). Dust levels were elevated during manual sorting and screening of waste and high levels of endotoxin and ß-(1-3) glucan were observed during all practices involving the movement of waste. A significant correlation was observed between the personal dust levels and personal endotoxin concentrations (r=0.783, p<0.05) and that personal inhalable dust concentration may be a valuable predictor for personal endotoxin concentration in the sites studied. CONCLUSIONS: Workers at composting sites are exposed to high levels of bacterial endotoxin consistent with adverse respiratory outcomes even though in most cases, their personal dust exposure is below the suggested regulatory levels. Dose-response data for the biological components present in the dust encountered at composting sites are not well established at this time and site operators should adopt precautionary measures when assessing and managing these potential risks.

A randomised comparison of SurePath liquid-based cytology and conventional smear cytology in a colposcopy clinic setting.

OBJECTIVE: The objective of this study was to compare the sensitivity of cervical cytology using conventional smears and SurePath liquid-based cytology (LBC). DESIGN: Prospective randomised evaluation of diagnostic test. SETTING: A single institution colposcopy clinic. POPULATION: Women attending first visit colposcopy appointments were offered entry into the study. METHODS: Cervical cytology samples from 913 women of age 16-75 years were randomly processed as SurePath LBC or conventional smears. Conventional smears were taken for 453 women and a SurePath LBC taken for 451 women. Cytology results were correlated with colposcopic findings and histology from colposcopic biopsies, treatment and follow up. MAIN OUTCOME MEASURES: To compare the sensitivity of SurePath LBC and conventional smears for histologically proven abnormality. Other outcome measures include a comparison of their sensitivity for high-grade abnormalities and their satisfactory rate. RESULTS: Accounting for all randomised samples, there was a trend towards improved sensitivity for SurePath LBC (79.1 versus 73.7%, P = 0.1). However, excluding unsatisfactory cytology (and samples not taken) eliminated this trend; the sensitivity for both LBC and conventional smears for any epithelial abnormality was 81%. With a threshold of atypical squamous cells of uncertain significance (ASC-US), both SurePath LBC and conventional smears had a sensitivity of 92% for high-grade lesions. SurePath LBC was less likely to be reported as unsatisfactory (2.7 versus 9.1%, P < 0.0001). CONCLUSIONS: In this context, with a threshold of ASC-US, both SurePath LBC and conventional smears offer high sensitivity for the detection of CIN2/3, but SurePath LBC is less likely to be reported as unsatisfactory.

Cellular transfer and AFM imaging of cancer cells using Bioimprint.

A technique for permanently capturing a replica impression of biological cells has been developed to facilitate analysis using nanometer resolution imaging tools, namely the atomic force microscope (AFM). The method, termed Bioimprint, creates a permanent cell 'footprint' in a non-biohazardous Poly (dimethylsiloxane) (PDMS) polymer composite. The transfer of nanometer scale biological information is presented as an alternative imaging technique at a resolution beyond that of optical microscopy. By transferring cell topology into a rigid medium more suited for AFM imaging, many of the limitations associated with scanning of biological specimens can be overcome. Potential for this technique is demonstrated by analyzing Bioimprint replicas created from human endometrial cancer cells. The high resolution transfer of this process is further detailed by imaging membrane morphological structures consistent with exocytosis. The integration of soft lithography to replicate biological materials presents an enhanced method for the study of biological systems at the nanoscale.

The PKZ1 recombination mutation assay: a sensitive assay for low dose studies.

The majority of mutation studies are performed at high doses of DNA damaging agents due to the insensitivity of most mutation assays. Extrapolation using a linear no-threshold (LNT) dose response model is then used to estimate the extent of possible DNA damage at lower doses. There is increasing evidence to suggest that the LNT model may not be correct at low doses of at least some DNA damaging agents. The pKZ1 in vivo and in vitro recombination assays have proven to be very sensitive for detection of changes in chromosomal inversion in lymphoid tissue in response to low doses of DNA damaging agents. Non-linear dose response curves for chromosomal inversion as an end-point have been identified at low doses of DNA damaging agents using this assay. Here, we review the inversion results obtained to date with the pKZ1 assays and discuss their suitability for low dose studies.

Does the density of lymphatic vascular space invasion affect the prognosis of stage Ib and IIA node negative carcinoma of the cervix?

Lymphatic vascular space invasion (LVSI) has been noted as a poor prognostic factor in many tumors. In some studies of carcinoma of the cervix, LVSI has been demonstrated to be independent of other prognostic factors. The aim of this study is to evaluate if, by a simple quantitative technique, the density of lymphatic invasion could be correlated with the risk of recurrence in node negative early stage carcinoma of the cervix. We analyzed the pathology and clinical course of 71 consecutive patients with stage IB and IIA carcinoma of the cervix treated primarily by radical hysterectomy and pelvic lymphadenectomy. All cases had negative nodes and adequate surgical margins. There were 67 patients suitable for evaluation. Tumour type, grade, stage and the dimensions of the tumor were recorded. The density of LVSI was categorized as absent (45%), mild (15%), moderate (33%) or severe (7%) depending on the number of lymphatic vascular spaces involved per high power field in the worst affected slide. The patients were followed for 2-8(1/2) years with a mean follow up of 4 years and 2 months. There were 13 recurrences and 7 deaths. All recurrences occurred in less than 2 years after surgery. The risk of recurrence was 40% for patients with extensive LVSI, 32% for moderate, 30% for mild and 3% if LVSI was absent. Only the presence of LVSI was associated with an increased risk of recurrence. The density of lymphatic invasion as represented by the number of lymphatic spaces occupied on the worst histological slide offered no further clinically useful information.

The pre-operative identification of low-risk endometrialcancer: an audit of women treated in the South Island of New Zealand 1998-2000.

OBJECTIVE: To determine whether pre-operative investigations identify a group of patients with low-risk endometrial cancer, who do not require tertiary referral for surgical staging or pelvic radiotherapy. DESIGN: Retrospective chart review. SETTING: South Island of New Zealand gynaecological oncology services. SAMPLE: One hundred and forty consecutive patients with newly diagnosed endometrial cancer from 1988 to 2000. METHODS: The results of preoperative investigations were compared with the final pathology. MAIN OUTCOME MEASURES: Correlation of preoperative investigations with low risk disease. For the purpose of the study, women with grade 1 or 2 endometrioid tumours confined to the uterine body and less than 50% myometrial invasion were considered to have low risk disease. RESULTS: In total, 50 women had low risk disease. Only 53% of patients with grade 1 tumours on initial biopsy had low risk disease. Women who had a grade 1 tumour at biopsy and, an ultrasound report with an endometrial thickness of less than 20 mm, and no evidence of myometrial invasion, cervical involvement or adnexal metastasis had a 76% chance of having low risk disease. CONCLUSION: We were unable to accurately define the low risk group from pre-operative assessment.

Gene transfer to ovine corneal endothelium.

PURPOSE: Modification of a donor cornea by gene therapy has potential to modulate irreversible rejection, the major cause of corneal graft failure. The sheep is a useful model for the human in this respect, as ovine endothelial cells are amitotic. The aim of the study was to investigate the ability of various non-viral and viral agents to transfer a reporter gene to ovine corneal endothelium. METHODS: The non-viral agents Transfectin-10, Transfectin-20, Transfectin-50, SuperFect, Effectene and CLONfectin were used to deliver the reporter gene, Escherichia coli lacZ, to ovine corneal endothelium in vitro. A Herpes simplex virus-1 and an adenoviral vector each encoding E. coli lacZ were similarly tested. Infected corneas were organ-cutured for up to 7 days in vitro to allow transfection efficiency, duration of gene expression and toxicity attributable to each vector to be compared. RESULTS: Scattered single or clusters of endothelial cells expressing the reporter gene were observed after transfection with CLONfectin, Transfectin-10, Transfectin-20 and Transfectin-50. SuperFect and Effectene were virtually ineffective. At best, the absolute number of infected cells per endothelial monolayer after 3 or 7 days of organ culture was estimated as < 0.01%. The Herpes simplex virus-1 vector also failed to transduce ovine corneal endothelium efficiently. In contrast, transfection rates of up to 70% of endothelial cells were observed with the adenoviral vector. CONCLUSION: Non-viral vectors and Herpes simplex virus-1 are unlikely to be suitable for gene therapy of corneal endothelium, because the efficiency of transfection is low compared with the rates achieved with adenoviral vectors.

The impact of the National Cervical Screening Programme on the presentation of cancer of the cervix in Canterbury.

AIMS: To assess the impact of the introduction of the National Cervical Screening Programme (NCSP) on the presentation of women with cervical cancer to Christchurch Women's Hospital (CWH), a tertiary referral center in Canterbury. METHOD: A retrospective review of the hospital notes of patients diagnosed with invasive cervical cancer from January 1988 to December 1990 and from January 1997 to December 1999. The patient characteristics, mode of presentation and staging during these two three-year study periods were compared. RESULTS: There was a 21% decline in the number of cases of invasive cervical cancer treated at CWH from the Canterbury region from the first to the second study period. There was also a significant clinical down staging at presentation and a significant increase in women presenting with asymptomatic smear detected disease (p=0.004). These changes were less marked in women older than 50 years. There was no change in the relative proportions of different histological types. CONCLUSION: The results reflect a positive impact on the presentation of cervical cancer associated with the introduction of the NCSP in Canterbury. A significant decrease in mortality from cervical cancer in the region can be anticipated. Further emphasis needs to be placed on prevention of cervical cancer in women over 50 years of age.