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As at present only a long‐term follow‐up can fully determine whether monoclonal gammapathies of undetermined significance (MGUS) will evolve into multiple myeloma (MM), this study attempted to identify other variables connected with the amount of monoclonal component (MC), generally considered as the most reliable marker of malignant evolution. Thirty‐four MGUS subjects showing a high MC (≥ 15.0 g/l) but without clinical evidence of MM (MGUS group b), were characterized for their phenotypic and genotypic profile by comparing them either with 40 MM patients or with 24 subjects affected by a benign form of monoclonal gammapathy (MGUS group a) according to the standard criteria. In addition to the usual laboratory markers, the levels of expression of a panel of CD membrane subsets were measured on B and T lymphocytes. Also, the serum level of the p53 mutant protein and the structural alterations of the c‐myc oncogene were evaluated. The results show that for MGUS group b patients, an increased M‐protein was accompanied by significantly increased levels of peripheral blood CD3(+) T cells and oncogenetic aberrations in c‐myc. Since a high serum MC level seems to indicate a greater likelihood of malignant transformation for MGUS patients, these findings suggest that this relationship may be a result of the concomitant alterations observed at a phenotypic and genotypic level. Such alterations may be potentially useful as surrogate markers for the transition of benign to malignant (MM) plasma cell dyscrasia.

Flow cytometric DNA content and proliferative kinetic markers, S-phase fraction (SPF) and thymidine labeling index (TLI), were evaluated in 68 patients with endometrial carcinoma. A high rate of aneuploid tumors was detected (48.4%); median values of SPF and TLI were 6.4 and 6.2, respectively. No significant relationship emerged between ploidy status and proliferative markers in respect to clinical and pathological variables. Aneuploid tumors had a higher recurrence rate than diploid tumors (21.8% vs 9.6%), but the difference was not statistically significant. According to the median value of both kinetic markers, the study population was divided into low and high-risk, where DFS was 100% and 71.4%, respectively (p = 0.05). Furthermore, high-TLI tumors (6.2) had a significantly worse DFS (75.4%) than low-TLI (100%) only among patients assigned to Stage I of the disease, regardless of other pathological variables. At multivariate analysis myometrial invasion resulted as an independent and significant factor. Flow cytometric ploidy analysis was useless as a predictive biological parameter and did not add any further prognostic information to the pathologic variables. SPF and TLI values could indicate a subset of women with unexpected poor outcome in a group of patients generally considered at low-risk, i.e. Stage I. If further investigation confirms these data, it could prove useful for therapeutic planning in endometrial cancer patients. At the present time, pathological and clinical factors are still the most reliable predictive parameters.

Models describing progression in the genetic derangement of glial tumors have shown chromosomal loss and gain occurring most frequently in high-grade lesions, suggesting that identification of these aberrations may be prognostically significant. In this study, Fluorescence in situ hybridization (FISH) has been used to determine, and to confirm, loss and gain of chromosomes 1, 8, 10, 12 and 17, in formalin-fixed, paraffin-embedded brain biopsy tissue taken from 60 brain gliomas submitted to surgical resection or stereotactic biopsy. FISH analysis may be a valuable adjunct to histological grading. The results showed that this molecular cytogenetic technique is an important clinical and experimental tool that provides new insight on genetic alterations, confirming gain and loss of genetic material that occurs at the initiation and progression of human glioma. Our data suggests that potentially useful prognostic information may be obtained through this approach. Monosomy 10 was the most statistically significant negative predictor of patient survival, showing a significant correlation with the histological grading.

Characterization of the biopathologic events underlying the early steps of breast carcinogenesis may have a dramatic impact on reducing breast cancer mortality. Genes involved in breast tumorigenesis are localized on chromosomes 1 and 17, and numeric aberrations of these chromosomes have been correlated with breast cancer tumorigenesis and progression. According to the field cancerization hypothesis, specific chromosome aberrations may be present in breast cancer and in normal-appearing adjacent tissue. The latter changes reflect the genomic damage that follows longterm carcinogenic exposure and precede the morphologically detectable neoplastic transformation. We hypothesize that detection of these aberrations in benign breast epithelium may provide a tool for molecular risk assessment.Using fluorescence in situ hybridization with centromere-specific probes, we determined the status of chromosomes 1 and 17 in fresh imprints of 28 samples of primary tumors and 54 samples of their surrounding uninvolved parenchyma taken from patients undergoing operations for breast carcinoma. Ten contralateral breast biopsy specimens collected from patients with previous breast carcinoma were also evaluated as a surrogate of a high-risk group to rule out the hypothesis that chromosomal aneusomy in tumor-adjacent tissue could be related to a paracrine effect of the primary tumor. Ten samples of benign breast tissue taken from patients at low risk were used as controls to define tolerance limits for aneusomy definition.Using threshold values of 40% of signal loss and 13% of signal gain to define chromosome aneusomy (ie, mean + 3 SDs of the control group signals), we found the following: 1) almost all primary breast tumors were aneusomic for chromosomes 1 and 17; 2) primary breast tumor and adjacent uninvolved parenchyma shared the same pattern of chromosomes 1 and 17 aneusomy in 66.7% of patients; and 3) chromosomes 1 and 17 aneusomies in contralateral benign breast samples from high-risk patients were not different from those in primary breast tumor or adjacent tissue samples.These results suggest that chromosomes 1 and 17 aneusomy may represent an intermediate biomarker of breast tumorigenesis potentially useful to detect patients at high risk of breast carcinoma who may benefit from preventive interventions.

The present study is the first to evaluate serum levels of vascular endothelial growth factor (VEGF) in B-cell chronic lymphocytic leukaemia (CLL). All 68 B-cell CLL patients and 31 control subjects analysed had detectable serum levels of VEGF, with no statistically significant difference between two proups. An aberrant increase of circulating levels of VEGF was found in only 17.6% of cases. B-cell CLL patients whose serum VEGF levels were higher than the median (i.e. 194.8 pg/ml) or 75th percentile (i.e. 288.5 pg/ml) values were more frequently at an advanced clinical stage. In contrast, no correlation with other clinico-biological features representative of either tumour mass [bone marrow (BM) histology, peripheral blood (PB) lymphocytosis, beta-2 microglobulin (beta-2m), LDH, interleukin-6 (IL-6)] or disease-progression (DP) [lymphocyte doubling time (LDT)] was found. Serum levels of VEGF predicted the risk of DP in early CLL. Among 41 patients in Binet stage A, progression-free survival (PFS) was significantly shorter in those patients whose VEGF serum concentrations were above the median value. Interestingly, characteristics of stage A patients stratified according to the median value of VEGF were similar with respect to many clinico-biological features, thus suggesting a possible independent prognostic role for such a marker. Finally, when added to the Rai subclassification, VEGF serum levels identified two groups with different PFS within stages I-II. We conclude that increased serum levels of VEGF can be considered useful for predicting the risk of DP and add prognostic information to the Rai subclassification of stage A CLL.

To determine the role of serum levels of IL-6 and p53 mutant protein as well as of c-myc proto-oncogene alterations: a) in discriminating between benign (MGUS) and malignant Plasma cell dyscrasias (Multiple and Microsecreting Myeloma, Plasmocytoma); b) in monitoring the clinical course of malignant forms of this disease.Eighty-eight patients affected by Plasma cell dyscrasias (58 MGUS, 24 MM and 6 PLC) entered this study. Using commercially available ELISA kits, serum levels of IL-6 and p53 have been determined in all the patients. In addition, a selected group of patients (n = 30) was also analyzed for structural c-myc gene alterations by Southern blot technique.The results show that, conversely from p53 protein, IL-6 and c-myc gene may represent useful diagnostic markers for discriminating benign from malignant forms of Plasma cell dyscrasia. On the contrary, preliminary findings of the same work indicate a potential role for the mutant p53 protein in monitoring the response to chemotherapy of patients affected by MM or PLM.Overall, these data suggest that the combined use of IL-6, p53 and c-myc may provide a new approach for a more rational management of Plasma cell dyscrasia patients.

CD27, a transmembrane homodimer belonging to the nerve growth factor (NGF) receptor superfamily, is typically expressed on leukemic CD5+ cells in B-cell chronic lymphocytic leukemia (CLL) and found in soluble form in the serum of CLL patients. Therefore, we investigated clinico-biological implications of increased serum levels of sCD27 in an unselected series of B-CLL patients.Serum CD27 (sCD27) levels were determined at the time of diagnosis in 82 previously untreated B-CLL patients using a sandwich enzyme-linked immunoassay (ELISA). Results were correlated with either clinico-hematological or biological features. Finally, quantitative flow cytometric analyses of membrane CD27 (mCD27) expression were carried out on peripheral blood (PB) cells of 22 B-CLL patients and 5 healthy controls, respectively.CD27 was found to be expressed on the surface of both resting normal and leukemic B cells. sCD27 levels were significantly higher in B-CLL patients (median value 2150 U/mL) than in healthy controls (median value 220 U/mL) (p0.0001). There was a close relationship between sCD27 and soluble TNF-alpha, another molecule belonging to the NGF receptor superfamily. Changes in sCD27 level correlated with clinical stage, beta 2 microglobulin and LDH.These findings indicate that sCD27 is a reliable marker of tumor mass in B-CLL. Its potential prognostic value should be tested in prospective studies.

Using a commercially available, competitive ELISA kit based on a polyclonal anti-interleukin-10 antibody, serum interleukin-10 (IL-10) levels were quantified in samples of different groups of patients: 20 healthy controls (CTR), 11 monoclonal gammopathies of uncertain significance (MGUS), 17 multiple myelomas (MM), 10 cancer patients (CANCER), 13 cancer patients + MGUS (MGUS-CA) and 7 MGUS patients after surgical removal of concomitant cancer (MGUS-SRCC). Results show significant differences of both the median levels of IL-10 and the monoclonal component (MC) in CTR, MGUS and MM (patients with increasing concentrations in the mentioned order). The IL-10 levels found in the three groups of cancer patients showed serum levels higher than those observed in the controls. Moreover, the surgical cancer removal was related to an IL-10 decrease. A higly significant correlation between serum IL-10 levels and the corresponding MC was also found in the MM-bearing patients and to a lesser extent, in MGUS patients, indicating that serum IL-10 is parallel to the amount of the activated clone causing the monoclonal gammopathy. Since human myeloma lines, cultured in vitro may release significant amounts of IL-10, the data presented support the hypothesis that serum IL-10, measured in myelomatous patients may, at least in part, derive from the activated clone causing the monoclonal gammopathy.

Antibodies against phospholipid antigens (APA) have been demonstrated in idiopathic thrombocytopenic purpura (ITP), but their clinical and pathogenetic significance has remained elusive. In this study we analyzed the prevalence and clinical features of ITP patients with elevated APA. In addition, we prospectively evaluated APA levels after treatment with corticosteroids and compared them with platelet-associated immunoglobulin (PAIgG) titers. We studied 149 patients with newly diagnosed ITP. Of these, 78 had a platelet count less than 50 x 10(9)/L and received an initial treatment with oral prednisone (PDN). In 71 asymptomatic cases with platelet counts between 50 x 10(9)/L and 120 x 10(9)/L, no therapy was scheduled. However, in five of them, the platelet count fell below 50 x 10(9)/L after more than 12 months; these patients were treated with PDN. Tests for APA included the measurement of anticardiolipin antibodies (ACA) with a solid-phase immunoassay and the detection of the lupus-like anticoagulant (LA) activity with coagulation tests that included kaolin-clotting time, dilute Russel's Viper venom time, activated partial thromboplastin time (aPTT), and dilute aPTT. Controls consisted of 174 apparently healthy subjects. Either LA or elevated ACA was seen in 69 patients (46.3%) at diagnosis. LA and ACA were both elevated in 24 cases (16.1% of the overall patient population and 34.8% of patients with high APA concentrations). No correlation was found between LA ratio values and ACA-IgG or -IgM titers, or between ACA-IgG and ACA-IgM levels. The presence of these antibodies was not associated with sex, age, platelet count, or the severity of hemorrhages. PAIgG was detected in 106 of 127 cases (83%). Again, no relationship was observed with clinical parameters or with APA levels. However, all cases with elevated APA also had increased PAIgG. With regard to the clinical course, we were not able to detect any significant difference between patients with normal and elevated APA. An initial complete response to prednisone treatment was observed in 43 of 83 cases (51.8%), with 13 (15.7%) achieving a prolonged complete remission. APA levels were not significantly modified after PDN therapy and on relapse. We conclude that APA positivity is a common finding in patients with ITP and does not select a category with different clinical features. APA levels are not influenced by immunosuppressive therapy with steroids and are not related to the activity of the disease. Therefore, we do not support a role for APA in the pathogenesis of ITP.

Fifty-four gastric biopsies and their relative gastric juices were analyzed for the presence of Helicobacter pylori with both cultural and microscopic methods. Thirty-one samples were positive and twenty-three were negative. These data were therefore employed as references for the subsequent comparisons. Furthermore, the gastric juices were later tested to establish the urea concentration and the pH level. In addition, the sediment obtained after centrifugation was microscopically observed for the possible presence of other bacterial flora in the sample (unstained smears). The urease test on the bioptic specimens has also been evaluated. The presence of H pylori was strictly related to urea levels of less than 15 mg/dl and pH less than 3.5. Furthermore, H pylori was generally not associated with the presence of other bacterial flora (only 1 out of 12 samples). The latter instead, was almost exclusively present in high pH samples (with the exception of one). On the basis of these results, a simple diagnostic scheme was constructed to identify carrier subjects. All patients (14/14) with urea levels of more than 15 mg/dl were found to be negative as well as those presenting a pH of more than 3.5 (7/9) or evidence of other bacteria in the juices (8/9). The remaining subjects (30/31 or 29/31, respectively) presented H pylori in the gastric juices. The final classification was 96.3% (or 94.4%) correct.

Two acute phase reactants, four cytokines, five soluble factors and lymphocyte subpopulations have been simultaneously evaluated in 16 subjects before and closely after the HIV-Ab seroconversion time. The same variables have also been determined in 50 HIV-Ab-negative high risk subjects, in 36 CDC II-III and in 30 CDC IV patients, utilizing a mixed longitudinal epidemiological model. The results show significant variations of few parameters in the early phases (increase: sCD8, beta-2-Microglobulin, sIL-2R, sCD23, Neopterin, IFN-alpha; decrease: CD4+ lymphocytes). In the course of the disease, many others parameters progressively increase (IFN-tau, IL-4, IL-6, acid-alpha 1-glycoprotein, alpha 1-antitrypsin) or decrease (B- and T-lymphocytes). Ferritin, in particular, highly increases only in CDC IV stage. These data may be useful to monitor patients during the entire course of their disease and to suggest the time elapsed from seroconversion.

We report the results of a randomized clinical trial of two different coricosteroids (prednisone versus deflazacort) in patients affected by autoimmune thrombocytopenic purpura (ATP). We have evaluated the efficacy of the two steroids on platelet count, antiplatelet antibodies, lymphocyte subsets and the occurrence of side effects. Twenty-seven patients were evaluable: 13 were treated with PDN and 14 with DFC. After 24 weeks of treatment, 4/12 (33%), subjects treated with PDN were refractory while complete responses were obtained in 2/12 (17%) and partial responses in 6/12 (50%). Among patients treated with DFC, 4/11 (36%) were considered as refractory, 2/11 (18%) had a complete response and 5/11 (46%) a partial response. A statistically significant decrease of antiplatelet antibodies was recorded in both groups after 4 weeks of therapy, but only in subjects receiving PDN did the reduction last until the 24th week. We observed an increase of T lymphocyte subsets (CD3, CD2, CD4, CD8) in absolute number, due to an increase in circulating lymphocytes, after 4 weeks. No substantial modifications were observed in these populations regarding the percentage or the CD4/CD8 ratio. After 24 weeks, 91% (10/11) of patients treated with PDN presented an increase of body weight and 1 had a stable increase in blood pressure. Among the subjects treated with DFC, 64% (7/11) showed an increase of body weight after the same follow-up. In conclusion, no difference was observed the two steroids studied.

"Giant cell" production by C. albicans cultured in the presence of substances belonging to the pentitol family is described in this study. After two days of incubation, at pentitol concentrations ranging between 0.1 and 20 mg%, the production of very large, frequently multibudding cells became evident. Their number increased with time, in some cases reaching 20% of the culture cells and their size could exceed 25 mcm. Comparisons with other sugar/alcohols and with other Candida spp. are presented. A possible explanation of this observation is consistent with an accumulation of pentitol catabolites in the yeast cytoplasm followed by an increased osmotic strength and cell swelling.

36 young women, aged 15-36, on oral combined contraceptives, were observed for a period of 3-24 months to study changes in lipid metabolism. Triglycerides serum concentrations and cholesterol levels were normal, except in 2 patients; total lipidemia and glycemia also were normal. These results demonstrate that oral contraceptives do not alter lipid metabolism. Modifications of this nature are sometimes found in older women, possibly because forms of lipid metabolism disorders can be latent in older age.

Two cases of benzodiazepine-induced thrombocytopenia are described. Drug-dependent antibodies with antiplatelet activity of the IgG class were detected by inhibition of complement-dependent hemolysis. This technique permitted also the demonstration of cross-reactivity of these antibodies with different benzodiazepines.