In vitro and in vivo models for the evaluation of new inhibitors of human steroid sulfatase, devoid of residual estrogenic activity.

The goal of our research project is to develop a new class of orally active drugs, estrone sulfatase inhibitors, for the treatment of estrogen-dependent (receptor positive) breast cancer. Several compounds were synthesized and their pharmacological potencies explored. Based on encouraging preliminary results, three of them, TX 1299, TX 1492 and TX 1506 were further studied in vitro as well as in vivo. They proved to be strong inhibitors of estrone sulfatase when measured on the whole human JEG-3 choriocarcinoma and MCF-7 breast cancer cells and their IC(50)s found to be in the range of known standard inhibitors. Their residual estrogenic activity was checked as negative in the test of induction of alkaline phosphatase (APase) activity in whole human endometrial adenocarcinoma Ishikawa cells. In addition, their effect on aromatase activity in JEG-3 cells was also examined, since the goal of inhibiting both sulfatase and aromatase activities appears very attractive. However, it has been unsuccessful so far. Then, in vivo potencies of TX 1299, the lead compound in our chemical series, were evaluated in comparison with 6,6,7-COUMATE, a non-steroidal standard, in two different rat models and by oral route. First, the absence of any residual estrogenic activity for these compounds was checked in the uterotrophic model in prepubescent female rats. Second, antiuterotrophic activity in adult ovariectomized rat supplemented with estrone sulfate (E(1)S), showed that both compounds were potent inhibitors, the power of TX 1299 relative to 6,6,7-COUMATE being around 80%. This assay was combined with uterine sulfatase level determination and confirmed the complete inhibition of this enzyme within the target organ. Preliminary studies indicated that other non-steroid compounds in the Théramex series were potent in vitro and in vivo inhibitors of estrone sulfatase in rats and further studies are in progress.

Small cell carcinoma of the esophagus.

Small cell carcinoma of the esophagus (SCEC) is a rare tumor with its particular biologic features, distinct from squamous and glandular carcinoma of esophagus. Although initial symptoms can be similar (with metastatic dissemination and paraneoplastic syndromes at presentation more frequent in SCEC), differences in age and sex distribution, tumor location, radiological and endoscopic findings, clinical course and prognosis have been observed between SCEC and other esophageal malignancies. SCEC should be considered a systemic disease, and by analogy to bronchogenic small cell carcinoma, multimodality approach including chemotherapy is recommended. In patients with limited disease, irradiation or surgery should be offered after induction chemotherapy to manage local disease. However, optimal treatment schedule has not yet been defined. In the future, registration of all SCEC cases and careful analysis of prognostic factors in the larger multi-institutional series could contribute to further progress in treatment outcome.

Interstitial brachytherapy for carcinomas of the lower lip. Results of treatment.

From 1973 to 1988 47 patients with previously untreated T1 and T2 squamous cell carcinomas of the lower lip received a definitive course of interstitial brachytherapy by iridium 192 wires. The disease stage was T1 in 21 cases (44.7%) and T2 in 26, and N0 in all cases except 2 of N1. Radiation therapy dose ranged between 6000 and 8000 cGy. Local control was obtained in 44 patients (93.6%). Treatment failure in the neck was observed in 3 patients (6.7%). The 5- and 10-year actuarial disease-free survival rates were 92% and 85%, respectively. A surgical salvage was attempted in 3 patients, with postoperative definitive control of the disease in 2. The 10-year actuarial overall survival was 95%. The incidence of complications was acceptable (10.6% of mucosal necrosis). An excellent or good cosmetic result was obtained in 91.7% of patients.

Daily low-dose carboplatin and standard radiotherapy in unresectable head and neck and lung cancers: a pilot study.

A total of 14 patients with locally advanced and unresectable head and neck (SCCHN) or non small cell lung cancer were treated with a definitive course of radiation therapy with conventional fractionation and 30 mg/m2 carboplatin (CBDCA) given daily as an i.v. infusion during the 1st, 3rd, 5th and 7th weeks of the combined treatment. The planned tumor dose of at least 7000 cGy was reached in all SCCHN patients except 1 (6600 cGy). The 2 NSCLC patients received 6320 and 5980 cGy, respectively. The planned total CBDCA-dose of 600 mg/m2 was administered in all patients. No treatment delays were required in 10 patients. Interruptions for severe mucositis or myelosuppression occurred in 4 patients (28.6%), but in no case did the delay exceed 1 week. Complete response was obtained in 8 patients (57.1%); 7 of the 12 with SCCHN and 1 of the 2 with NSCLC. The other 6 patients achieved a partial response. Granulocytopenia of WHO grade 3 occurred in 1 patient; apart from vomiting and mucositis, toxicities above grade 2 were not observed.

[Role of radiotherapy in the combined treatment of invasive cancer of the vulva]. / Ruolo della radioterapia nel trattamento integrato del carcinoma invasivo della vulva.

The authors review the literature on the different combinations of radiotherapy with chemotherapy and surgery with the aim of giving a state of art on the role of combined multimodality treatment of invasive vulvar carcinoma. From the data of the recent literature it appears that radiation integrated with surgery and chemotherapy can play an important role in reducing the risk of postoperative locoregional failure in patients with advanced primary or nodal disease and avoiding exenteration in patients with disease involving the anus or proximal urethra. This integrated multimodality therapy is a promising approach in the treatment of invasive vulvar carcinoma but further exploration in a larger number of patients is needed before giving consolidated data applicable in routine oncological clinical practice.

Transitory and activity-dependent expression of neurogranin in olfactory bulb tufted cells during mouse postnatal development.

Neurogranin (Ng) is a brain-specific postsynaptic calmodulin-binding protein involved in synaptic activity-dependent plasticity. In the adult olfactory bulb (OB), Ng is expressed by a large population of GABAergic interneurons in the granule cell layer. We show here that, during postnatal development, Ng is also expressed by OB neurons in the superficial external plexiform layer (sEPL) and glomerular layer (GL). These Ng-positive neurons display morphological and neurochemical features of superficial and external tufted cells. Ng expression in these cells is transient during OB development: few elements express Ng at postnatal day (P) 5, increasing in number and reaching a peak at P10, then progressively decreasing. At P30, Ng is rarely detectable in these neurons. Ng expression in developing tufted cells is also modulated at the cellular level: at earlier stages, Ng labeling is distributed throughout the cell body and dendritic arborization in the GL, but, at P20, when the glomerular circuits are fully matured, Ng becomes restricted to the soma and proximal portion of tufted cell apical dendrites. We show that olfactory deprivation at early postnatal stages induces a strong increase in Ng-positive tufted cells from P10 to P20, whereas no changes have been observed following olfactory deprivation in adult mice. These findings demonstrate that Ng expression in sEPL-GL is restricted to developmental stages and indicate its activity-dependent regulation in a time window critical for glomerular circuit development, suggesting a role for Ng in maturation and dendritic remodeling of tufted cells.

Expression and localization of the calmodulin-binding protein neurogranin in the adult mouse olfactory bulb.

Neurogranin (Ng) is a brain-specific postsynaptic protein involved in activity-dependent synaptic plasticity through modulation of Ca(2+)/calmodulin (CaM)-dependent signal transduction in neurons. In this study, using biochemical and immunohistochemical approaches, we demonstrate Ng expression in the adult mouse olfactory bulb (OB), the first relay station in odor information processing. We show that Ng is principally associated with the granule cell layer (GCL), which is composed of granule cell inhibitory interneurons. This cell type is continuously renewed during adult life and plays a key role in OB circuits, integrating and modulating the activity of mitral/tufted cells. Our results indicate that Ng localizes in the soma and dendrites of a defined subpopulation of mature GABAergic granule cells, enriched in the deep portion of the GCL. Ng-immunopositive cells largely coexpress the Ca(+)/CaM-dependent kinase IV (CaMKIV), a downstream protein of CaM signaling cascade, whereas no colocalization was observed between Ng and the calcium-binding protein calretinin. Finally, we demonstrate that adult neurogenesis contributes to the Ng-expressing population, with more newly generated Ng-positive cells integrated in the deep GCL. Together, these results provide a new specific neurochemical marker to identify a subpopulation of olfactory granule cells and suggest possible functional implications for Ng in OB plasticity mechanisms.

[Survival and recurrence of endometrial adenocarcinoma]. / Sopravvivenza e recidive dell'adenocarcinoma dell'endometrio.

444 endometrial carcinomas were retrospectively reviewed. Disease-free survival (DFS) was 83.6% at five years and 80.5% at ten years. 349 T1 cancers were treated by TAH-BSO and post-operative irradiation; 22.6% of them were over-staged. DFS ranged from 100% to 91% in 270 low-risk T1 patients, dropping to 84% in intermediate risk patients and to 44% in high-risk patients. DFS was 93% and 66% in the patients overstaged as pT2 and pT3, respectively; it was 82% for ovarian spread and 45% for pelvic peritoneum/parametrium invasion. T2-T3 patients were treated with preoperative irradiation and bilateral hysterectomy. DFS was 90.5% vs. 72.2% and 2/2 vs. 3/7 when the lesion was understaged. The results of irradiation were similar to literature data, but 53% of patients (all in poor general conditions and/or very old) died from associated diseases. Relapses were observed in 13.6% of cases (62/444 patients); 29 (6%) were local recurrences and 33 (7.6%) were distant metastases, which correlated well with recurrences and 33 (7.6%) were distant metastases, which correlated well with risk factors. Severe complication (G3) were observed in 6.3% of cases.

Concomitant radiotherapy and daily low-dose carboplatin in locally advanced, unresectable head and neck cancer. Definitive results of a phase I-II study.

The combination of daily low-dose carboplatin and radiotherapy was studied in 55 patients with inoperable head and neck cancer. All patients were planned to receive 70 Gy plus carboplatin i.v. daily, 45-60 min before radiotherapy. A starting schedule of 30 mg/m2 on days 1 through 5, weeks 1, 3, 5 and 7 was administered to 17 patients; an escalating daily dose, up to 55 mg/m2, was given to 38 additional patients. Up to a daily dose of 45 mg/m2, only 4.4% of the patients developed grade 3 leukopenia; on the contrary, grade 3 and 4 leukopenia was seen in 62.5% of patients receiving 50 mg/m2 or more. Mucositis was the major nonhaematologic toxicity and seemed to be dose-dependent. At the end of the loco-regional treatment there were 33 (61.1%) CR and 17 PR; the most effective total carboplatin dose seemed to be 40-45 mg/m2. After surgical salvage the number of CRs increased to 37 (68.5%). One- and 2-year loco-regional control rates were 64% and 53% respectively. One- and 2-year actuarial survival rates were 71% and 53% respectively; the corresponding rates of disease-free survival were 60% and 43%. There was a strong correlation nodal status and both survival and disease-free survival.