It is commonly believed that tumor cells treated with anticancer agents, chemotherapy and/or radiation, die by apoptosis and that tumors which do not undergo apoptosis are resistant to treatment. In this study, we investigated the molecular basis underlying cisplatin cytotoxicity in the murine teratocarcinoma F9 cell line to see whether irradiation enhances cisplatin-induced cytotoxicity. We compared the apoptosis induced by chemo and/or radiotherapy with other cellular effects such as cell survival, clonogenic capability, cell cycle perturbation, expression of p53 and p53-related mRNAs, and necrosis. When combined with radiation, a clear additive cytotoxic effect of cisplatin was demonstrated. We found that both cisplatin and radiation induced cell death, but the level of induced apoptosis was low and there was no correlation with the results of the clonogenic assays: we noted a difference between cytotoxic effects in the clonogenic assay and the extent of apoptosis by fluorescence-activated cell sorter analysis, suggesting that cell killing reflected not only apoptosis but also cell cycle arrest, and that apoptosis, cell kinetics and clonogenicity suppression were independent processes.

BACKGROUND:Pathologic staging and grading are the most important prognostic factors in prostatic cancer. Unfortunately, pathologic staging needs to be evaluated by surgical procedures; moreover, the proposed grading systems are largely based upon subjective histopathologic evaluations. In recent years, these problems have been approached with flow cytometry (FCM). The present study evaluates the significance of DNA ploidy of prostatic carcinoma assessed by FCM.MATERIALS AND METHODS:Microscopic slides from 132 core needle biopsies and 22 surgical specimens of prostatic carcinoma were reviewed and classified according to Gleason's score modified by Epstein et al. Formalin-fixed, paraffin-embedded tissue samples were prepared for FCM evaluation using standard techniques. Adequate histograms were obtained from 113 biopsies (85.6%) and from all the surgical specimens (100%).RESULTS:Among the 113 biopsy specimens, a statistically significant correlation was found between DNA ploidy and Epstein's grading, since high-grade neoplasms accounted for 40.38% of non-diploid cases and only for 21.33% of diploid cases (chi square = 5.8; p = 0.05). Moreover, tetraploid tumors were defined as a separate FCM category with an absolute prevalence (65.52%) of non-high grade neoplasms (chi square = 5.9; p = 0.05). Although not submitted to statistical analysis, data collected from surgical specimens showed similar distribution.CONCLUSIONS:DNA ploidy assessment by FCM is a viable procedure in the majority of needle biopsy tissue samples of prostatic carcinoma; it may produce prognostic parameters with greater objectivity than conventional histologic grading.

BACKGROUND DNA flow cytometry of hepatocellular carcinoma (HCC) cells has been investigated in many studies, but, to the best of our knowledge, there are no data on DNA analysis of cirrhotic parenchyma around the HCC. In this study, cell kinetics and ploidy of parenchymal cells around HCC were performed to ascertain if this would predict the possibility of recurrence in the cirrhotic areas. METHODS The DNA content of 93 cases of HCC and of cirrhotic liver around the tumor nodules was analyzed by flow cytometry. Ploidy and proliferative index of HCC and cirrhotic liver were compared with macroscopic, histologic, and clinical features of each case and linked with the behavior of these tumors. Survival curves were assessed according to the Kaplan-Meier method. A multivariate analysis based on Cox proportional hazards regression model was performed on cases of diploid cirrhosis cells in which the S-phase fraction was evaluable. RESULTS The univariate analysis of survival suggested significant roles for age, number of intrahepatic nodules, Edmondson-Steiner's classification, portal invasion, vascular invasion, presence of necrosis, hepatitis B surface antigen, α-fetoprotein, Child's score, ploidy, and S-phase fraction of HCC cells. The DNA analysis of the cirrhotic cells showed that polyploidy was dramatically reduced in patients with HCC, compared with normal hepatocytes, and aneuploid clones were present among diploid cells. High S-phase fraction of cirrhotic cells and Child-Pugh classification were the strongest independent parameters affecting the tumor behavior in this study. CONCLUSIONS The results of this study suggest that S-phase fraction of cirrhotic liver parenchyma may be employed as a new parameter in the prognostic evaluation of HCC patients. Cancer 1996;78:1195-1202.

Nuclear DNA content was determined on paraffin blocks of 28 neuroblastic tumors retrieved from Surgical Pathology files. In 26 cases cytofluorimetric analysis for DNA content was satisfactory. 8 tumor were diploid, 12 were aneuploid and 6 tetraploid. All but one of the children with diploid neuroblastoma (NB) died after a survival ranging from 14 to 32 months. The only surviving child had a Stage I thoracic ganglioneuroblastoma (GNB) and was alive after 27 months. Conversely 11 of 12 patients with aneuploid tumors were alive with prolonged follow-up. Two children are alive after a period exceeding 146 months. The only exception in this group of aneuploid tumors was a child with Stage IV neuroblastoma of the adrenal gland with pleuro pulmonary metastasis who died after 6 months. In the group with tetraploid tumors 3 patients died (2 children with Stage III and IV died early while the third, with thoracic Stage II GNB, survived for 72 months). Two patients with tetraploid GNB Stage II and III are alive 65 and 72 months after diagnosis respectively. Another child with Stage IV adrenal NB is alive after 14 months but with metastatic spread to bones, bone marrow and lymph nodes. Cox analysis of the cases demonstrated ploidy as an independent prognostic factor. These results are in agreement with other molecular analysis linking near-ploidy of Neuroblastic tumors to poor prognosis. Ploidy, as detected by flow cytometry for nuclear DNA content, may represent an important prognostic factor in Neuroblastic tumors. The advantage of flow cytometry over other techniques of molecular analysis is represented by the simple methodology suitable for fixed and paraffin embedded tissue, even on retrieval material.

A case of enteropathy associated T-cell lymphoma (EATCL) in a 62-year-old female with a previous history of coeliac disease, complicated during the clinical course by massive blood and tissue eosinophilia is described. The patient's serum contained a factor capable of stimulating the in vitro growth of eosinophilic colonies (CFU-Eo), that was absent in the serum of normal donors. We suggest that such factor was Interleukin-5 (IL-5), as indicated by the presence in the monoclonal tumor T cells of IL-5 encoding mRNA, usually absent in the normal enterocytes of the jejunum.

By flow cytometric analysis of nuclear DNA content it is possible to demonstrate the cellular clones that have a quantity of DNA higher or lower in comparison with that of diploid nuclei. The possibility of measuring DNA ploidy by flow cytometry provides a simple way to asses triploidy and tetraploidy in placental tissue. In this investigation, are reported the results of the DNA analysis in a series of 540 consecutive spontaneous abortions or suction curettage material obtained after intrauterine death, which was detected by ultrasound examination. At microscopy 351 (65.0%) cases showed feature of normal placentas, 147 (27.2%) of hydropic abortus, 27 (5.0%) of partial hydatidiform mole and 15 (2.8%) of complete hydatidiform mole. DNA flow cytometric analysis showed triploidy in 38 cases (7.0%) and tetraploidy in 12 cases (2.2%). Comparing microscopic feature and ploidy, 12 normal placental specimens were triploid, 6 were tetraploid. Nine hydropic abortuses had a triploid DNA content, 5 a tetraploid DNA content. In the group of partial hydatidiform moles triploidy has been found in 14 cases, whereas tetraploidy in one case and diploidy in 12 cases. Twelve complete hydatidiform moles showed a diploid DNA content and 3 a triploid one. The incidence of poliploid pregnancies has been higher among young patients. In most cases the poliploid pregnancy was the first pregnancy of the patient. DNA flow cytometric analysis is a simple and useful mean to state etiology of many spontaneous abortion, even when histology is negative. This analysis must be associated with morphology in this field.(ABSTRACT TRUNCATED AT 250 WORDS)

In the medico-legal practice differential diagnosis between spontaneous and non-spontaneous abortion is important because causes of pregnancy wastage are often obscure and, moreover, spontaneous abortion is more common than accidental or voluntary. In all the cases in which the cause of abortion is not otherwise detectable and especially in cases of discovery of fetal adnexa, it is necessary to investigate genetic causes. Recently, DNA flow cytometric analysis has been applied in determining the genetic causes of spontaneous abortions. Among karyotypic abnormalities, flow cytometric analysis on paraffin embedded material can detect only polyploidies (triploidy and tetraploidy). Trisomies, monosomies and structural anomalies cannot be detected. In our study we tried to establish whether flow cytometry could be useful in determining the genetic cause of spontaneous abortions, in the lack of any other detectable cause. Histologic examination and flow cytometric analysis were performed on a series of 395 consecutive spontaneous abortions. Histologic examination allowed the detection of a molar pattern in about 9% of cases. DNA flow cytometric analysis showed diploidy in 346 (87.59%) cases, triploidy in 37 (9.36%) cases and tetraploidy in 12 (3.03%) cases. Combined microscopic and flow cytometric analysis revealed abnormalities in 17.5% of cases. A non-diploid pattern is more frequent in molar cases (P < 0.001). Flow cytometry seems to be interesting in forensic pathology, as it allows the detection of some frequent genetic abnormalities in dead tissues and cells, when other techniques are no longer practicable.

INTERLEUKIN-2 (IL-i?) stimulates immunity by acting on specific IL-2 cell surface receptors, expressed by activated but not resting immune cells [ 11. It has been demonstrated that lymphocytes may produce a soluble form of IL-2 receptors(sIL-2r) [2]. We have reported increased sIL-2r serum levels in lung cancer Table 1. Median values of sIL-2r in healthy subjects and in lung cancer patients

A review of the literature concerning the analysis of nuclear DNA content by flow cytometry is made and it is compared with own experience. The advantages and the limits of this technique are examined. The practical problems in the interpretation of the histograms and the value of the measurements are discussed. The importance of this analysis in diagnosis and in staging of many tumors and the clinical involvements are emphasized. In many tumors DNA ploidy represents a new independent prognostic variable that is useful to separate the cases with a potential worse behaviour in an early stage, when other classic parameter are not available. It is also stressed the importance of flow cytometric DNA analysis of tumor cells. This is made on the same tissues that pathologists use for histopathological diagnosis and the results are important in many cases for a diagnostic accuracy.

Flow cytometric analysis of DNA content was done on 133 primary squamous cell carcinomas of the larynx. Overall, 76 tumors (57.1%) were not diploid (aneuploid or tetraploid). The DNA index (DI) was calculated and grouped into three levels. Fifty-seven cases had a DI in a range of 1.85 to 2.15 (44 diploid and 13 tetraploid). The cases were grouped in relation to ploidy, proliferative index, and the tumor-node-metastasis (TNM) system. Every group was analyzed with respect to survival rate. Ploidy was related to histocytologic grade. In well-differentiated tumors (G1) survival rates at 48 months were 41.7% in diploid cases and 27.7% in nondiploid ones (relative risk [RR], 2.01; 95% confidence interval [CI], 0.89, 4.52). In NO cases that underwent surgery, survival rates at 48 months were, respectively, 81.8% and 49.2% (RR, 5.07; 95% CI, 0.76, 33.93). These results suggest that ploidy may be a new independent parameter of prognosis in squamous cell carcinoma of the larynx. This is useful in clinical practice because it allows the clinician to recognize those cases with poorer prognosis among the early tumors at a stage where other prognostic parameters are not yet available.

Plasma cell acid phosphatase (AP) activity and thymidine labeling index (LI%) were evaluated concomitantly in 52 patients with monoclonal gammopathies. AP score, percentage of AP positive plasma cells, and LI% were significantly higher in 26 patients with multiple myeloma (MM) at the time of diagnosis than in 11 monoclonal gammopathy of undetermined significance (MGUS) and eight smoldering myeloma (SM) patients. LI% had the highest statistical correlation with disease status. A 1% cutoff could clearly separate the patients with progressive MM compared to those with stable disease (SM-MGUS) (P less than .001). There was a significant overall correlation between the AP score and LI% (P less than .005). Since LI% is a recognized powerful prognostic factor, this correlation suggests that the AP score can also be a reliable test predicting patient survival duration. In addition, we identified a subgroup of IgG MM patients with very high tumor mass who had a low LI% but a high AP score. This was associated with very poor patient survival and indicated the discrete prognostic importance of AP score in this subgroup with low LI%. Thus, both the LI% and AP score can be recommended as helpful clinical tests in patients with monoclonal gammopathies.