Influence of soil properties and phosphate addition on arsenic uptake from polluted soils by velvetgrass (Holcus lanatus).

Four kinds of soil material were used in a pot experiment with velvetgrass (Holcus lanatus). Two unpolluted soils: sand (S) and loam (L) were spiked with sodium arsenite (As II) and arsenate (As V), to obtain total arsenic (As) concentrations of 500 mg As kg(-1). Two other soils (ZS I, ZS III), containing 3320 and 5350 mg As kg(-1), were collected from Zloty Stok where gold and arsenic ores were mined and processed for several centuries. The effects of phosphate addition on plants growth and As uptake were investigated. Phosphate was applied to soils in the form of NH4H2PO4 at the rate 0.2 g P/kg. Average concentrations of arsenic in the shoots of velvetgrass grown in spiked soils S and L without P amendment were in the range 18-210 mg As kg(-1) d.wt., whereas those in plants grown on ZS I and ZS II soils were considerably lower, and varied in the range 11-52 mg As kg(-1) d.wt. The addition of phosphate caused a significant increase in plant biomass and therefore the total amounts of As taken up by plants, however, the differences in As concentrations in the shoots of velvetgrass amended and non-amended with phosphate were not statistically significant.

BRCA1-positive breast cancers in young women from Poland.

We identified 4316 unselected incident cases of early-onset breast cancers (<51 ears of age at diagnosis) in 18 Polish hospitals between 1996 and 2003. We were able to obtain a blood sample for DNA analysis from 3472 of these (80.4%). All cases were tested for the presence of three founder mutations in BRCA1. The proportion of cases with a BRCA1 mutation was 5.7%. The hereditary proportions were higher than this for women with breast cancer diagnosed before age 40 (9%), for women with cancer of medullary or atypical medullary histology (28%), for those with bilateral cancer (29%) or with a family history of breast or ovarian cancer (13%). It is reasonable to offer genetic testing to women with early-onset breast cancer in Poland.

BRCA2 germline mutations in male breast cancer patients in the Polish population.

Breast cancer is a rare disease in men. Germ-line mutations in BRCA2 and androgen receptor (AR) genes are thought to be responsible for a proportion of male breast cancer cases. The present study was performed on a series of 37 consenting patients not selected for family history of breast/ovarian cancer. The entire coding region of the BRCA2 gene and two exons of the AR gene were analyzed for germ-line mutations to evaluate the association between BRCA2 and AR genes and male breast cancer in Poland. We identified four frameshift mutations (11%) in exons 10, 11, 17 and 18, two of them were novel: 6495del3insC and 8457insA. Three missense unclassified variants (8%) of the BRCA2 gene were also identified. The frequencies of missense alterations were examined in a set of 200 chromosomes. No alteration of the AR gene was found. We did not observe much difference in clinicopathological features between carriers and non-carriers of BRCA2 mutations. Five of 37 patients (14%) had a family history of breast cancer, in one first- or second-degree relative, among the latter was one mutation carrier. The results of this study suggest that germ-line BRCA2 mutations account for rather small proportion of male breast cancer in Poland.

Genetically modified tumour vaccines (GMTV) in melanoma clinical trials.

Since melanoma is a model immunogenic malignancy incurable in the disseminated phase of its natural course different immunotherapeutic approaches are tested in clinical trials. A number of tumour vaccines genetically modified (GMTV), with various immunostimulatory factors, are tested in phase I/II clinical trials. These factors include cytokines, tumour antigens (TA), costimulatory molecules or HLA antigens. We have designed a novel, mixed auto/allogeneic cellular melanoma vaccine modified with the IL-6 and the sIL-6R genes. Preclinical studies in a mouse model demonstrated that the IL-6/sIL-6R based vaccine is able to elicit efficient anti-tumour responses, mediated by CD8+ and NK cells, which resulted in inhibition of the tumour growth, metastases formation and prolonged survival of the animals treated. Irradiation of vaccine cells does not only lead to their sterilisation but also causes increased secretion of exogenous IL-6 and sIL-6R. Since January 1996 we have vaccinated more than one hundred metastatic melanoma patients. Promising clinical results (22% CR+PR, 32% SD) and the evidence of immune responses in the vaccinated patients have prompted us to design a phase III clinical trial which is to be open in 2000.

Expression of interleukin-6, interleukin-6 receptor, and glycoprotein 130 correlates with good prognoses for patients with breast carcinoma.

BACKGROUND: Interleukin-6 (IL-6) is secreted by normal epithelial breast cells but not by oncogene-transformed cells. Interleukin-6 is able to inhibit growth of breast carcinoma cells in culture. Interleukin-6 exerts its activity via two receptor subunits, IL-6R and glycoprotein 130 (gp130). The expression of these receptor subunits in breast tumors has been studied, but there are no previous reports of their prognostic significance, to the authors' knowledge. METHODS: mRNA of IL-6, IL-6R, and gp130 was studied in 75 tumor samples obtained from breast carcinoma patients. Patients were followed for a maximum of 71 months (median follow-up, 61 months; 60 patients were followed for a minimum of 5 years or died during the observation period). Prognostic factors were analyzed in univariate and multivariate analysis. RESULTS: mRNA specific to IL-6, IL-6R, and gp130 was detected in 57%, 53%, and 71% of breast carcinoma tissues, respectively. Expression was strongly correlated with earlier stages of the disease. In univariate analysis, expression of IL-6 and its receptor subunits proved to be a positive prognostic factor for overall survival (OS) and disease free survival (DFS). IL-6R and gp130 expression were good independent prognostic factors for OS. The 5-year OS of all patients was 66%. The 5-year OS in IL-6, IL-6R, and gp130 positive groups was 95%, 94%, and 90%, respectively, whereas in negative groups it was 26%, 31%, and 9%, respectively. CONCLUSIONS: Expression of IL-6, IL-6R, and gp130 in breast carcinoma tissue is associated with earlier stages of the disease. In advanced stages, expression of IL-6 and its receptor subunits predicts better prognosis.

New protein and PCR markers RAK for diagnosis, prognosis and surgery guidance for breast cancer.

Breast cancer antigens RAK-p120, -p42, -p25 were detected in 100% of breast cancer cases tested (71 cases). Only 10% of adjacent tissue cases tested positive for all three cancer antigens, and 17.5% of the cases tested positive for two antigens only. Eighty-five percent of histologically normal breast tissue samples, isolated either from breast cancer patients or patients with advanced fibrocystic disease, tested RAK-negative, with the exception of low expression of p25, observed in some patients. Polymerase chain reaction (PCR) with HIV-1 gp 41-derived primers revealed cancer-associated DNA fragments of similar size (140 bp) as in HIV-1 genome. Fifty-four percent of cancer adjacent tissues, and 50% of malignancy-free breast tissue samples, tested PCR-negative. It is suggested that genetic predisposition to cancer may be associated with the presence of RAK genes, while expression of RAK antigens marks an already ongoing process of malignant changes.

Helicobacter pylori from duodenal ulcer patients expresses inducible nitric oxide synthase immunoreactivity in vivo and in vitro.

Helicobacter pylori is a major pathogenic factor in gastritis, ulcer disease and gastric cancer. Helicobacter pylori associated gastritis is of complex pathogenesis which is only partially elucidated. In the present study we investigated by immunohistochemical BSA method the expression of inducible nitric oxide synthase in the gastric wall and in vitro in Helicobacter pylori. In the gastric wall nitric oxide synthase immunoreactivity was found in macrophages, in endothelial cells and in neural elements. In Helicobacter pylori nitric oxide synthase was also expressed. In conclusion, we postulate a possibility of a direct toxic effect of Helicobacter pylori to gastric epithelial cells through the nitric oxide radical.

Diagnostic evaluation of cancer antigens RAK .1. Cervical and ovarian cancer.

Cancer antigens RAK-p120, p42, and p25, which exhibit biological, immunological and molecular similarity to the proteins expressed by Human Immunodeficiency Virus 1 (HIV-1), were found in 47 of 47 tested cases of serous adenocarcinoma of the ovary, and 45 of 45 tested cases of squamous carcinoma of the cervix. Normal ovary and cervix did not express antigens RAK. High molecular weight protein (RAK-p160) was detected in the blood of over 61% of ovarian and 72% of cervical cancer patients, and in 14.3% of healthy women with family history of breast and/or gynecological cancer. Antigens RAK might represent new diagnostic markers.

Soluble interleukin 6 receptor is biologically active in vivo.

A cancer cell transfection model was used to evaluate biological activity of soluble IL-6 receptor (sIL-6R) in vivo. B-78 melanoma cells were stably transfected with cDNAs encoding human IL-6, murine sIL-6R and human leukaemia inhibitory factor (LIF). Control and transfected cells were intravenously (i.v.) and/or subcutaneously (s.c.) injected into B57BL/6 x C3H or SCID mice. Whereas B-78 cells formed tumours and lung metastasis in injected animals, transfected animals, transfected cells showed greatly reduced tumour and metastasis formation. Transfection of IL-6, sIL-6R or LIF had similar protective effects. The combination of IL-6 and sIL-6R was most effective. Kinetic analysis demonstrated a 3 week lag period between formation of tumours by B-78 cells and the combination of B-78 cells transfected with IL-6 and sIL-6R. No such lag phase was seen when B-78-IL-6 or B-78-IL-6 or B-78-sIL-6R were injected alone. These results indicate that IL-6 alone exhibits a different quality of activity when compared to the IL-6-soluble receptor complex. Our results demonstrate for the first time that sIL-6R is a biologically active molecule in vivo.