Feulgen cytometry in simultaneous endometrial and ovarian carcinoma

The modal DNA value was measured in six patients with simultaneous endometrial and ovarian carcinoma by Feulgen static cytometry. One patient with low-grade endometrial and ovarian carcinoma manifested diploid indices at both sites. Another patient demonstrated aneuploid ovarian carcinoma and diploid endometrial carcinoma, indicating that these were separate neoplasms. The remaining four patients with Stage III disease had aneuploid endometrial and ovarian carcinomas with identical DNA indices. These data support a single neoplastic process with metastasis in the latter four patients. There was good correlation with the clinicopathologic impression on the likelihood of synchronous primaries versus metastatic neoplasms. It was concluded that DNA analysis is a useful adjunct in assessing the probability that spatially separate neoplasms represent metastasis.     

采用Feuloen细胞光度法测定法探讨鼻咽低分化癌...

Nuclear area and DNA content of the normal columnar cells and cancer cells which were small, fusiform, irregular, vesicular-nuclear, giant malformed-nuclear and giant vesicular-nuclear were measured by Leitz MPV-III microscope photometer with HP-85 microcomputer in 32 poorly differentiated nasopharyngeal carcinoma (NPC) and 5 chronic nasopharyngitis sections. Based on different DNA contents and distribution patterns, it was divided into 4 cancer cell populations: small, fusiform and/or irregular, vesicular-nuclear and giant tumor cell populations. It should be noted that the nuclear DNA pattern of vesicular-nuclear cancer cell population had pathognomonic characteristics. The nuclear unit DNA content ratio of vesicular-nuclear cancer cells to lymphocytes was under 0.4. This cancer cell population was sensitive to irradiation. If the biological characteristics of these four cancer cell populations could be clarified not only in nuclear DNA pattern but also in biochemistry at molecular level, it will be possible to design the different treatment trials on the different cancer cell populations, then resulting in better prognosis of NPC patients.     

Flow cytometry for detection and evaluation of urinary bladder carcinoma.

Flow Cytometry (FCM) DNA assays of bladder irrigation specimens are now recognized as a clinically useful and reliable means of detecting and monitoring carcinoma of the bladder. This technique, which identifies carcinoma by the presence of an aneuploid population of cells, can be carried out on specimens obtained in an outpatient or hospital setting and is easily performed in any medium-sized laboratory. It is most sensitive to superficial and high grade tumors. Overall, nearly 80% of superficial carcinomas of bladder will have positive flow cytometry, comparing very favorably with conventional cytology. Until now, the widest clinical application of FCM has been in monitoring the conservative treatment of stage 0-1 flat and papillary carcinomas, but newly developed dual parameter measurements are capable of quantifying proliferative activity, oncogene expression, growth factor receptors, and other cellular features that may better characterize the biologic potential of these tumors and can be expected to aid in the selection and timing of treatment.     

Flow cytometry evaluation of erythroid dysplasia in patients with myelodysplastic syndrome.

Erythroid dysplasia is the pathologic hallmark of myelodysplastic syndromes (MDS). To develop a quantitative flow-cytometry approach to its evaluation, we analyzed the expression of CD71, CD105, cytosolic H-ferritin (HF), cytosolic L-ferritin (LF) and mitochondrial ferritin (MtF) in erythroblasts from 104 MDS patients, 69 pathologic control patients and 19 healthy subjects. Six-parameter, 4-color flow cytometry was employed, and data were expressed as mean fluorescence intensity. Compared with pathologic and healthy controls, MDS patients had higher expression of HF (P < 0.001) and CD105 (P < 0.001), and lower expression of CD71 (P < 0.001). MtF was specifically detected in MDS with ringed sideroblasts, and there was a close relationship between its expression and Prussian blue staining (r = 0.89, P < 0.001). In vitro cultures of myelodysplastic hematopoietic progenitors showed that both HF and MtF were expressed at a very early stage of erythroid differentiation, and that MtF expression is specifically related to mitochondrial iron loading. A classification function based on expression levels of HF, CD71 and CD105 allowed us to correctly classify > 95% of MDS patients. This flow-cytometry approach provides an accurate quantitative evaluation of erythroid dysplasia and allows a reliable diagnosis of sideroblastic anemia, and may therefore be a useful tool in the work-up of patients with MDS.     

Flow cytometry evaluation of erythroid and myeloid dysplasia in patients with myelodysplastic syndrome.

The purpose of this study was to develop a flow cytometric approach to the evaluation of marrow dysplasia in myelodysplastic syndromes (MDS). We first studied a cohort of 103 MDS patients as well as 46 pathological and healthy controls. Flow cytometry data were expressed as percentage of positive cells. Analysis of erythroid cells showed higher proportions of immature cells (P < 0.001) and decreased levels of CD71 expression on nucleated red cells (P = 0.02) in MDS. Analysis of myeloid cells showed lower proportions of CD10+ and higher proportions of CD56+ granulocytes (P < 0.001), and increased ratios of immature to mature cells (P = 0.007). Since no single immunophenotype could accurately differentiate MDS from other conditions, we used discriminant analysis for generating erythroid and myeloid classification functions using combinations of immunophenotypic parameters. These functions were prospectively validated in a testing cohort of 69 MDS patients and 46 pathological controls. A diagnosis of MDS was obtained in 60/69 cases (87%). No false-positive results were noticed among controls. Significant correlations between values of these functions and both degree of morphological dysplasia and the International Prognostic Scoring System were found. These findings indicate that flow cytometry evaluation of marrow dysplasia is feasible and may be useful in the work-up of individual MDS patients.     

流式细胞术用于良恶性胸腔积液鉴别诊断的研究--DNA倍体分析和突变型p53蛋白检测

目的研究流式细胞术DNA倍体和细胞周期分析、突变型p53检测用于良恶性胸腔积液鉴别诊断的临床可行性.方法 24例标本(12例恶性,12例良性)采用流式细胞术细胞内抗原检测的直接免疫荧光标记法检测胸腔积液细胞中表达突变型p53的阳性细胞百分率和p53表达的平均荧光强度,并且通过PI染色分析细胞倍型和周期分布.结果流式细胞术DNA倍体分析用于恶性胸腔积液诊断的敏感性为66.7%,特异性100%.倍体分析和细胞学检查联合应用能将诊断的敏感性提高到100%,特异性仍为100%.良性胸腔积液和恶性胸腔积液p53表达有显著性差异.以p53阳性细胞百分率＞90%为阳性标准,p53用于良恶性胸腔积液鉴别诊断的敏感性和特异性分别为83.3%和91.7%.联合p53阳性细胞百分率和DNA倍体分析对恶性胸腔积液诊断的敏感性能提高到91.7%,特异性91.7%.细胞周期SPF(S期时相百分比)在良恶性胸腔积液之间没有显著差异.结论流式细胞术DNA倍体分析和突变型p53检测可以作为临床良恶性胸腔积液鉴别诊断的辅助手段,结合细胞学检查更有意义.     

Flow cytometry analysis of DNA damage and the evaluation of cytotoxicity of alkylating agents

Monoclonal antibody (MAb) F7-26 generated against nitrogen mustard (HN2)-treated DNA (O.S. Frankfurt, Exp. Cell Res., 170: 369-380, 1987) reacted with regions of local DNA denaturation (distortion) induced by DNA alkylation. The relationship between immunoreactivity of cellular DNA with MAb F7-26 and cytotoxic effects of HN2, L-phenylalanine mustard (L-PAM), and 1,3-bis(2-chloroethyl)-1-nitrosourea was studied in HeLa S3 cultures. Cells were treated with drugs for 1 h and assessed for cell survival by colony formation assay and for DNA immunoreactivity by flow cytometry. Cells were fixed in ethanol, exposed to MAb, and stained with fluorescein-labeled anti-mouse immunoglobulin. Immunofluorescence (IF) intensity was measured on a flow cytometer. For each drug the cell killing and the binding of MAb to DNA appeared in the same dose ranges. A strong correlation (r = 0.96) between cell survival (log10 surviving fraction) and IF was observed when data for HN2, L-PAM, and 1,3-bis(2-chloroethyl)-1-nitrosourea were combined. This correlation was apparent in the range of 1-5 log10 cell killing. Enhancement of L-PAM cytotoxicity by buthionine sulfoximine (BSO) or hyperthermia was accompanied by a proportional increase of DNA immunoreactivity with MAb F7-26. The enhancement factors calculated from survival curves (a ratio of the dose decreasing cell survival by 1 log10 for L-PAM alone to that for L-PAM combined with modulating factor) were 1.67, 1.58, and 3.07 for BSO, hyperthermia, and BSO plus hyperthermia, respectively. For the same treatment regimens the enhancement factors calculated from drug dose-IF curves were 1.73, 1.34, and 3.79. A strong correlation between log10 surviving fraction and IF intensity (r = 0.93) was observed when data for L-PAM alone or L-PAM combined with BSO and/or hyperthermia were considered together in the range of 1-6 log10 cell killing. The cytotoxicity of alkylating agents and nitrosoureas and the effectiveness of factors modulating chemotherapeutic effects can be predicted by flow cytometry analysis of DNA immunoreactivity with MAb F7-26.     

Flow cytometry and interactive image cytometry in endometrial carcinoma. A comparative and prognostic study

Comparative DNA measurements were performed in 139 women with endometrial carcinoma using flow cytometry (FCM) and interactive image cytometry (ICM). Ploidy level and the percentage of S-phase cells were determined by FCM and ploidy level and the percentage of cells with a DNA content exceeding 2.5c, 3c, 4c and 5c, respectively, were calculated by ICM. The aim was to compare ploidy level obtained by the two methods and to evaluate the prognostic value of all the above-mentioned parameters. Recurrence or residual disease after completing treatment were used as end-points. Follow-up time was 18-48 months. An agreement was obtained in 85% of the cases as regards ploidy level, but in 15% the tumors were regarded as near-diploid by one method and as grossly aneuploid by the other. Both ploidy level (both methods) and S-phase rate (FCM) were correlated with histopathologic grade (p less than 0.001 and p less than 0.05, respectively). In univariate analysis, ploidy level (obtained either by FCM or ICM) correlated with recurrence rate, with a more favourable prognosis for near-diploid cases. When using multivariate models (Cox analysis) including clinical variables, ploidy level by FCM (but not ICM) was still significant as regards prognosis. In the multivariate analysis, S-phase fraction (as measured by FCM) also yielded independent prognostic information. In a separate analysis the proportion of cells with a DNA content greater than 5c gave independent prognostic information besides that of the S-phase fraction and ploidy level. We conclude that measurements of the percentage of cells exceeding 5c give prognostic information beyond the information obtained from flow cytometric determination of ploidy level and S-phase fraction.     

Flow cytometry in exfoliative cytology of malignant tumor

Exfoliative cells from 170 cases of malignant tumor and 70 cases of normal tissue were studied by flow cytometry. The results showed that 155/170 (91.2%) cases were positive and 15/170 (8.8%) were false-negative; 4/70 (6.0%) cases were false-positive. The authors consider that flow cytometry has achieved a diagnostic accuracy comparable to the conventional cytology. Flow cytometry is a useful supplementary method in the diagnosis of tumors.     

Flow cytometry: an overview.

Flow cytometry is a method of rapidly analyzing large numbers of cells as they flow in a liquid medium through a laser source. Flow cytometry provides valuable information regarding DNA content, RNA content, and the percentage of cells in the S phase of the cell cycle. This information can be used to predict the aggressiveness of many tumors, which has direct prognostic and diagnostic implications. Flow cytometry assists healthcare professionals in planning and implementing effective, appropriate, and timely interventions. Because flow cytometry is becoming more prevalent, nurses must be aware of its actual and potential applications. The data obtained from flow cytometry provide important information that can assist nurses in anticipating a particular regimen for specific tumors and in projecting the patient's clinical course. With this information, nurses can modify and individualize care plans. When used in conjunction with a patient's clinical presentation and other laboratory findings, flow cytometry has a direct impact on both treatment decisions and nursing interventions.     

Rapid detection of neoplastic cells in serous cavity effusions in children with flow cytometry immunophenotyping

The aim of the study was to evaluate the role of flow cytometric immunophenotyping (FCI) as a rapid diagnostic tool for pediatric malignancy in serous cavity effusions (SCEs). FCI results for 103 SCEs in a pediatric population were compared with retrospective clinical outcomes (RCOs). Among 41 patients assessed as having malignancies by RCO, 36 patients were diagnosed with lymphoma (n =25), acute myeloid leukemia (n =2), neuroblastoma (n =8) and retinoblastoma (n =1) by FCI, respectively. The sensitivity and specificity of FCI for detecting neoplastic cells were 87.8% and 98.4%, respectively. The concordance of FCI data with final diagnoses of the patients was 94.2%. FCI data for lymphoma was concordant with the final diagnosis in 89.3% of cases. When Hodgkin lymphoma was excluded, the overall correlation increased to 96.1%. FCI is a useful tool for rapid and reliable diagnosis of pediatric non-Hodgkin lymphoma in SCE samples and also to suggest the presence of non-lymphoid malignancies, especially neuroblastoma.     

Flow cytometry analysis of hematopoietic stem cells

Blood formation is sustained by a population of undiferentiated and metabolically quiescent hematopoietic stem cells (HSC) found in the bone marrow. HSC remain in the Go compartment of the cell cycle, are able to self-renew, and differentiate into progenitors of all hematopoietic lineages. A subset of hematopoietic cells suspected of harboring HSC express the cell surface antigen CD₃₄; CD₃₄+ purified fractions are enriched in colony-forming units, whereas negative fractions are depleted. CD₃₄+ cells obtained from either bone marrow or periphe-ral blood are increasingly used in bone marrow transplantation. The accurate enumeration of CD₃₄+ cells may be important for predicting the success of engraftment after transplantation, as it can assure that sufficient numbers of progentor cells remain in the intended transplant graft. For these reasons we developed a new method for the enumeration of CD₃₄+ cells in whole blood samples, that takes advantage of classic methods that are widely used in flow cytometry. However, a stem cell most accurately may refers to a biological function. Recent advances in flow cytometry may provide a fast and accurate way of measuring the presence and quality of HSCs.     

Flow cytometry of platelets for clinical analysis

Platelets are small, non-homogenous cells with distinctive surface features important to their essential role in hemostasis. The surface membrane is dynamic, and changes remarkably in lipid asymmetry and receptor expression on triggering of the activation process. There are also extensive and rapid intracellular changes in platelets as a result of biochemical activation through calcium fluxes, phospholipase activity, kinase activity, and phosphorylation mechanisms that lead to release of storage granule contents and generation of fast-acting prostaglandins, all in a matter of seconds after stimulation with a strong agonist. These characteristics make the platelet an interesting but difficult cell to study, and the explosion of knowledge over the last two decades has been fueled in large part by the application of flow cytometry techniques. Clinical applications of flow cytometry analysis of platelets have been pursued in individual specialized medical centers, but have not found widespread practice in clinical laboratories, mostly because of difficulties in standardization of techniques and the inherent biovariability in comparing normal to abnormal platelets. Despite these hurdles, it seems certain that flow cytometry analysis of platelets in pathological states will continue to evolve into more practical and robust procedures that will eventually become standard hematologic assays rather than specialized research tools.     

Flow cytometry of liver fine needle aspirates

Fine needle aspiration analysis of intraabdominal masses is increasing in frequency. This study was designed to evaluate whether flow cytometry could be of value in the cytopathology interpretation of such specimens. Over a 4-year time span, 129 consecutive liver fine needle aspirates were evaluated by flow cytometry for DNA content, ploidy, and proliferation rate. Only excess cells remaining in the needle after cytology samples were prepared were included in this study. Overall sensitivity was 75% and specificity was 94%. In addition, flow cytometry results were pivotal for at least two specimens in achieving the appropriate diagnosis. For these reasons, it was concluded that flow cytometry could be a valuable adjunctive technology to the cytopathologist in the interpretation of liver fine needle aspirates. © 1994 Wiley-Liss, Inc.     

Clinical and flow cytometry characteristics of malignant pleural effusions in patients after intracavitary administration of methylprednisolone acetate

Ten patients with recurrent pleural effusions due to advanced cancer were treated by intracavitary methylprednisolone acetate (Depo-Medrol [DM], Upjohn, Kalamazoo, MI). They received one to six courses of DM (median, three courses per patient) with doses ranging from 80 to 160 mg per course. Effusion cells were cryopreserved before and during DM installation for subsequent determination of ploidy by flow cytometry. Pleural effusion in all three patients with advanced breast cancer resolved and did not reaccumulate throughout follow-up for 11+, 10+, and 8+ months. Pleural effusion in a patient with metastatic gastric cancer and in two of four patients with adenocarcinoma of unknown origin partially resolved. Altogether six of ten patients (60%) subjectively and objectively benefited from this therapy. All patients tolerated the treatment well with no local or systemic side effects. Flow cytometry showed a reduction in ploidy of effusion cells in all three patients with breast cancer, from a peak mean channel of 6C to nearly 2C after therapy. Transient reduction of ploidy was seen also in the effusion of a patient with unknown primary tumor associated with clinical improvement. The clinical and laboratory data reported offers initial evidence that DM when instilled into the pleural cavity after incomplete thoracentesis may act as effective palliative therapy either alone or in combination with other anticancer agents.     

Flow cytometry in analysis of precancerous lesions of the esophagus

DNA content of the severe dysplasia cells in the esophageal epithelium was quantitatively analysed using flow cytometry and compared with those of the normal, mild dysplasia and cancer cells. The results showed that DNA index of the severe dysplasia cells was 1.27 +/- 0.11 and there was a significant difference between its value and those of the normal (1.0 +/- 0.02), mild dysplasia (1.01 +/- 0.03) and cancer cells (1.73 +/- 0.35). The DNA content of severe dysplasia cells lies between the normal and cancer cells. The hyperplasia degree of the severe dysplasia cells parallels their DNA contents. This study indicates that flow cytometry is a useful supplementary tool for diagnosis of the tumor.