Molecular expression of 17 beta hydroxysteroid dehydrogenase types in relation to their activity in intact human prostate cancer cells.

In the present study we have inspected estrogen metabolism in cultured human prostate cancer cells (LNCaP, DU145, PC3), in relation to the expression of mRNAs for different 17 beta hydroxysteroid dehydrogenase (17 beta HSD) enzymes (from 1 to 4). Using an intact cell analysis, we have compared precursor degradation and product formation after incubation of cells with physiological amounts of radioactive E2 or estrone (E1) for 24-72 h and subsequent reverse-phase high performance liquid chromatography analysis. The LNCaP and DU145 cells only partly converted E2 to E1 (26 and 13% at 72 h, respectively), giving rise to an appreciable production of E2 from E1 (nearly 20% in all cases). Conversely, PC3 cells revealed a massive E2 oxidation to E1 (up to 90% by 72 h) and a scant formation of E2 (<2%) from E1. In addition, an appreciable formation of 16 alpha OHE1 was seen in either PC3 (11%) or DU145 (5%) cells. respectively using E2 or E1 as precursor. All three cell lines exhibited marked amounts of 17 beta HSD4 mRNA species, whilst even greater amounts of 17 beta HSD2 transcript were found in PC3 cells only. No mRNA for either 17 beta HSD1 or 17 beta HSD3 could be detected in any cell line. The present evidence indicates that pathways of estrogen metabolism are distinctly governed in prostate cancer cells depending on their endocrine status, being associated with a differential expression of mRNA for different 17 beta HSD enzymes.

Product of aromatase activity in intact LNCaP and MCF-7 human cancer cells.

We investigated conversion rates of androgens to estrogens in cultured, hormone-responsive prostate (LNCaP) and breast (MCF-7) human cancer cells. For this purpose, we adopted an intact cell analysis, whereby cells were incubated for different incubation times in the presence of close-to-physiological (1 nM) or supraphysiological (1 microM) concentrations of labelled androgen precursors, i.e. testosterone (T) and androstenedione (delta4Ad). The aromatase activity, as measured by estrogen formation, was detected in LNCaP cells (0.5 pmol/ml), even though to a significantly lower extent than in MCF-7 cells (5.4 pmol/ml), using 1 microM T after 72 h incubation. Surprisingly, LNCaP cells displayed a much higher aromatase activity when T was used as a substrate with respect to delta4Ad. In either cell line, T transformation to delta4Ad was relatively low, attaining only 2.8% in LNCaP and 7.5% MCF-7 cells. However, T was mostly converted to conjugates (over 95%), glucuronides and some sulphates, in LNCaP cells, whereas it was only partly converted to sulphates (<10%) in MCF-7 cells. Aromatase activity seems to be inconsistent in LNCaP cells, being strongly affected by culture conditions, especially by fetal calf serum (FCS). Further studies should assess the regulation of aromatase expression by serum or growth factors in different human cancer cells, also using anti-aromatase and/or anti-estrogen compounds, in different culture conditions.

Central nervous system metastases in patients with ovarian carcinoma. A report of 23 cases and a literature review.

BACKGROUND: Central nervous system (CNS) involvement by ovarian carcinoma is rare. PATIENTS AND METHODS: From September 1982 to September 1994, 23 patients with CNS metastases from ovarian carcinoma were observed in our institution. RESULTS: Their median age at the time of CNS metastasis diagnosis was 59 years, and the median interval between diagnosis of ovarian cancer and documentation of the metastasis was 35 months. The most common symptoms related to CNS involvement were motor weakness, headache, seizures, dizziness and visual disturbances. One patient had meningeal carcinomatosis; 22 had parenchymal lesions (18 cerebral and 4 cerebellar). Nine patients had a single CNS lesion, and 13 had multiple metastatic sites. CNS was the only site of disease in 9 patients, while 8 had concomitant extraperitoneal dissemination. The median survival (MS) from diagnosis of cerebral metastases for the entire series was five months. Four patients were not treated (MS 3 months); 14 received radiotherapy (MS 5.5 months), and five underwent surgical resection of solitary metastases followed by radiotherapy (MS 17 months). Number of CNS lesions, extent of the disease at the time of CNS metastasis and treatment were the only factors which significantly affected survival CONCLUSIONS: The prognosis of patients with CNS metastasis from ovarian carcinoma appears poor. However, early diagnosis followed by multimodal treatment may result in significant palliation and improve overall survival in a selected group of patients.

Growth of LNCaP human prostate cancer cells is stimulated by estradiol via its own receptor.

We report that growth of LNCaP human prostate cancer cells is significantly stimulated (up to 120% above control) by physiological estradiol (E2) concentrations. This growth increase appears to be comparable to that induced by either testosterone or dihydrotestosterone, as also reported by others. This paper presents novel illustrative evidence for estrogen-binding proteins and messenger RNA transcripts in LNCaP cells. In fact, 1) the reverse transcriptase-polymerase chain reaction system documented normal messenger RNA for estrogen receptors (ER); 2) the radioligand binding assay allowed the detection of high affinity, reduced capacity binding sites in both soluble and nuclear cell fractions; and 3) the immunocytochemical analysis showed a consistently intensive staining for both ER and progesterone receptors. Compared to other human estrogen-responsive mammary cancer cells, MCF7 and ZR75-1, ER expression in LNCaP cells was not significantly lower, as shown by levels of the ER transcripts, number of sites per cell, or femtomoles per mg DNA as well as the percentage and intensity of immunocytochemical staining. A relative estimate of ER expression obtained by matching LNCaP with another human prostate cancer cell line, PC3, always displayed significantly and consistently higher levels in LNCaP cells. The detection of relatively high type I ER content in either cell compartment of LNCaP cells was paralleled by a highly intensive staining for progesterone receptors. In addition, evidence that the synthetic androgen R1881 did not compete for type I binding of E2 and that any E2-induced growth was completely reversed by the pure antiestrogen ICI-182,780, but unaffected by the antiandrogen Casodex, clearly suggests that the biological response of LNCaP cells to E2 is mediated via its own receptor.

Long-term peripheral neurotoxicity of cisplatin in patients with successfully treated epithelial ovarian cancer.

We evaluated clinically and neurophysiologically the immediate and long-term involvement of the peripheral nervous system in 22 selected patients with epithelial ovarian cancer successfully treated with DDP alone or in combination with non-neurotoxic drugs. While the motor nerves were unaffected, generally the involvement of sensory nerves was more severe at the examination performed several months after DDP discontinuation than at the evaluation performed after the "induction phase". We conclude that up to now the importance of long-term DDP-induced peripheral nerve damage has probably been underestimated. DDP-induced long-term damage is at least as severe as the immediate toxicity and, moreover, it is likely that complete recovery can occur, if ever, only years after DDP discontinuation.

Effect of isoprinosine on IL-2, IFN-gamma and IL-4 production in vivo and in vitro.

The effects of an immunopotentiating drug, isoprinosine, on the splenocytes of BALB/c mice to produce cytokines were investigated. Isoprinosine enhanced IL-2 production, upregulating the expression of IL-2 receptor in vitro. It also significantly increased the IFN-gamma secretion and decreased the IL-4 production in vivo. The significance of these findings in terms of immune regulation is discussed.

A sero-epidemiological survey of asymptomatic cases of Boutonneuse fever in western Sicily.

963 sera from contacts, persons from various localities, and blood donors were examined with a commercially produced kit for micro-immunofluorescence for the presence of antibodies to Rickettsia conorii. 10.6% of sera were serologically positive. The higher rates of positivity were observed in sera of contacts (19%) and persons from Mussomeli (20%) and Ustica (18.4%), the lower rates in blood donors from Palermo (3.5%). These results support the view that there is an occupational risk factor related to a rural environment.

[Seroimmunologic studies in boutonneuse fever. I. Evaluation of a commercial micro-immunofluorescence kit in the serodiagnosis of boutonneuse fever]. / Studi sieroimmunologici nella Febbre Bottonosa. I. Valutazione di un kit del commercio per micro-immunofluorescenza nella diagnostica sierologica della Febbre Bottonosa.

The diagnosis of Boutonneuse Fever usually depends on clinical evidence (summer occurrence, fever, tache noire at the site of the tick bite, in 30-70% of cases, erythemato -papular rash, prompt response to chloramphenicol or tetracycline treatment). Serological confirmation is difficult since the only diagnostic procedure currently feasible, the Weil-Felix test, is not specific. Other more specific diagnostic procedures (agglutination, complement fixation, ELISA, indirect immunofluorescence tests) are beyond the possibilities of most laboratories (antigens are not available from the market). In the present paper, results obtained with a new commercially produced kit for indirect immunofluorescence are reported. Sera from patients with infectious and non infectious diseases as well as Boutonneuse Fever (at various stages of illness, from 6 days to 12 months) were examined. Sera from blood donors were also included. Specificity and sensitivity were satisfactory as well as reproducibility of results. Some apparently false positivities must be related to the present epidemiological pattern in western Sicily, and namely to the incidence of asymptomatic cases of Boutonneuse Fever, as demonstrated by recent works.

[Preliminary results of the use of an ELISA test for the differential determination of IgG and specific IgM in toxoplasmosis serology]. / Primi risultati dell'impiego di un test ELISA per la determinazione differenziata delle IgG ed IgM specifiche nella sierologia della toxoplasmosi.

The diagnosis of acute toxoplasmosis usually depends on serology, since clinical and/or histological features are difficult and/or often misleading. IgM titers are the best indicators of infection acquired in the past two to four months. IgG titers are generally correlated with non-active infections as well as previous (symptomatic and/or asymptomatic) illness. In the present paper, results obtained with a new kit for the evaluation of IgG and specific-IgM by micro-ELISA technique, are reported. Optical density values in sera from: a) blood donors; b) miscellany group; c) suspected toxoplasmosis, were at various degree high for IgG. Few sera of the entire sample (in the group of suspected toxoplasmosis and one in miscellany) showed high optical density for specific IgM. Reading of results with spectrophotometer were in agreement with reading to the naked eye. Reproducibility was satisfactory. Unfortunately some sera positive in the first determinations gave equivocal results in successive proofs. For these reasons diagnosis of toxoplasmosis must be based upon the use of at least three tests.

[Further observations on the use of counterimmunoelectrophoresis (CIEP) on cellulose acetate membrane (Cellogel) in the diagnosis of visceral leishmaniasis (author's transl)]. / Ulteriori osservazioni sull'uso della controimmunoelettrosoresi (CIEP) su membrana di acetato di cellulosa (Cellogel) nella diagnostica a della leishmaniosi viscerale.

Technique of counterimmunoelectrophoresis (CIEP) on cellulose acetate membrane (Cellogel) is described for diagnosis of (human and canine) visceral leishmaniasis (VL). Various lots of antigen were grossly obtained from the liquid phase (10, 20 and 30 tubes respectively) of cultures of Leishmania donovani by repeated freezing and thawing. Sera from patients (and, in a few cases, dogs) with confirmed VL, other parasitic and non-parastic diseases (especially blood disorders and hepatosplenomegaly) and from blood donors were tested. Positive results were obtained in 91-93% (according to various lots) of the patients with VL. All sera from infected dogs gave positive results. No precipitin lines were detected in the control sera. Antigens from 30 (or 20 tubes) showed better results with regard to the evidence of the precipitin lines. Clarity of the precipitin bands appears to be in relation to: 1. the protein concentration of the antigens; 2. the antibody levels of the sera. CIEP on cellulose acetate membrane combine the features of good sensitivity, specificity and speed of performance and appear available for use also in epidemiological research.