SEARCH · PubMed Health

Results for “Flow Cytometry”

Explore indexed PubMed citations for clinical trials, systematic reviews and public health research. Read source abstracts and follow each citation to its original PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 685 records · Page 38Linked to original sources

PubMed2005

Variation in concentration of prion protein in the peripheral blood of patients with variant and sporadic Creutzfeldt-Jakob disease detected by dissociation enhanced lanthanide fluoroimmunoassay and flow cytometry.

BACKGROUND: A highly sensitive dissociation-enhanced lanthanide fluoroimmunoassay (DELFIA) and flow cytometry techniques have previously been developed and employed to characterize soluble cellular prion protein (PrP(c)) expression in whole blood and separated components from healthy adult blood donors. No previous studies with these techniques have evaluated the concentration and expression of PrP in the blood of patients with variant Creutzfeldt-Jakob disease (vCJD). STUDY DESIGN AND METHODS: For blood from vCJD patients, sporadic CJD (sCJD) patients, non-CJD neurological controls, and healthy adults, PrP(c) was measured by DELFIA and cell-associated PrP was measured by flow cytometry. RESULTS: DELFIA analysis identified a significant reduction in the concentration of PrP(c) in the whole blood of vCJD (p = 0.012) and non-CJD neurological patients (p = 0.0004) compared with healthy adults. A significant elevation was found in plasma PrP(c) in sCJD patients compared with healthy adult (p = 0.022) and neurological controls (p = 0.050). Flow cytometry found no significant differences between groups in expression of PrP on platelets and lymphocytes, nor in sensitivity of cellular PrP to proteinase K. Neurological controls show significantly less PrP on red cells than healthy adults. CONCLUSION: There are differences in free and cell-associated PrP found in blood of CJD patients and control groups, some of which might be useful with other tests in disease profiling as an aid to diagnoses.

Adult↗

PubMed1993

Assessment of the acrosomal status and viability of human spermatozoa simultaneously using flow cytometry.

Acrosomal status and viability were evaluated simultaneously on human spermatozoa using flow cytometry. Samples were divided into three aliquots and randomly assigned to one of three treatments: (i) cryopreservation; (ii) 10 microM calcium ionophore [A23187 in dimethylsulphoxide (DMSO)] or (iii) DMSO alone (control). Acrosomal status was evaluated using monoclonal antibodies recognizing MH61 and CD46, respectively. Fluorescein-conjugated goat anti-mouse immunoglobulin (IgG) was used as a second antibody. Sperm viability was assessed using Hoechst 33258 (H258) exclusion. The following factors were analysed: (i) the specificity of the monoclonal antibodies for the human acrosome; (ii) the relative effectiveness of flow cytometry and direct fluorescent microscopy scoring and (iii) the acrosomal status and viability of the control, ionophore-treated, and cryopreserved spermatozoa. Across all treatments, the MH61 and CD46 monoclonal antibodies resulted in acrosomal status values (acrosome-reacted/viable spermatozoa) which were not significantly different (P > 0.05): control, 1.0 +/- 0.3% and 1.5 +/- 0.6% (mean +/- SEM); A23187, 42.8 +/- 3.5% and 38.1 +/- 3.5%; cryopreserved, 8.2 +/- 2.0% and 9.9 +/- 1.3%; respectively. However, acrosomal status among treatments differed significantly (P < 0.01). Flow cytometric and direct fluorescent microscopy assessments were significantly correlated (r2 = 0.96, P < 0.01). These results indicate that flow cytometry, using an acrosome-specific monoclonal antibody and a supravital dye, provides an objective and efficient method to evaluate human sperm acrosomal and viability status simultaneously.

Acrosome↗

PubMed1995

Flow cytometry analysis of dual red blood cell populations after bone marrow transplantation.

Flow cytometry represents an alternative method to agglutination assays for the accurate quantification of mixed field populations of erythrocytes observed after bone marrow transplantation. Murine monoclonal antibodies directed against the blood group ABH antigens were selected and processed in order to prepare ready-to-use fluorescent reagents. Anti-A (NaM87-1F6; IgG3), anti-B (NaM9-2E11; IgG3) and anti-H (NaM19-7E11; IgM) were purified, labelled with fluorescein isothiocyanate, and used in a direct flow cytometry assay. Anti-A1 (NaM1-1C9; IgG3) was no longer active after FITC-labelling and then was used in an indirect assay. The agglutination was prevented by formaldehyde pretreatment of erythrocytes. Using artificially-made double populations of erythrocytes, measured values with mixtures of 1-100% of cells were very closely related to expected values, showing both the sensitivity and the accuracy of the method. From careful investigation of a series of bone-marrow transplanted patients, we conclude that engraftments could be demonstrated earlier by flow cytometry than by agglutination, because minor populations (1-10%) of cells could be determined accurately only with labelled reagents. In addition, the disappearance of the donor cells on a long-term follow-up of patients enabled an earlier detection of graft failure in one case. The proposed method provides appreciable help to follow engraftment in patients and may have more general applications for the study of other haemopoietic chimaeras.

ABO Blood-Group System↗

PubMed2001

HTPS flow cytometry: a novel platform for automated high throughput drug discovery and characterization.

The flow cytometer is unique among biomedical analysis instruments because it makes simultaneous and multiple optical measurements on individual cells or particles at high rates. High throughput flow cytometry represents a potentially important multifactorial approach for screening large combinatorial libraries of compounds. Limiting this approach has been the availability of instrumentation and methods in flow cytometry for automated sample handling on the scale required for drug discovery applications. Here, we describe an automated system in which a novel patented fluidics-based pharmacology platform, the HTPS (High Throughput Pharmacological System), is coupled to a flow cytometer using a recently described plug flow-coupling valve technology. Individual samples are aspirated sequentially from microplate wells and delivered to a flow cytometer for rapid multiparametric analysis. For primary screening to detect and quantify cell fluorescence in endpoint assays, a high-speed no-wash protocol enabled processing of 9-10 cell samples/min from 96-well microplates. In an alternate primary screening format, soluble receptor ligands were sampled from microplate wells at rates of 3-4 samples/minute and successfully assessed for the ability to elicit intracellular calcium responses. Experiments with fluorescent beads validated the accurate automated production by the HTPS of exponential and linear gradients of soluble compounds. This feature enabled rapid (2- to 3-min) characterization of the intracellular calcium dose response of myeloid cells to formyl peptide as well as the quantitative relationship between formyl peptide receptor occupancy and cell response. HTPS flow cytometry thus represents a powerful high throughput multifactorial approach to increase the efficiency with which novel bioresponse-modifying drugs may be identified and characterized.

Automation↗

PubMed1997

A comparison of cytogenetic studies and flow cytometry in breast carcinomas.

In this study, we compared genetic instability in 70 breast carcinomas analyzed by two different methods, cytogenetics and flow cytometry. This comparison showed that each method has its strengths and weaknesses. Flow cytometry detected aneuploidy in 60% of cases, but missed most of the cytogenetically near-diploid clones and clones with simple chromosomal changes. Cytogenetics revealed chromosomal abnormalities in 50% of the samples. Simple chromosomal changes and multiploidy were readily detected by this method, but some of the clones with a high DNA index by flow cytometry were missed. The two methods gave corresponding results in the majority of cases (54%). In 17 cases, both methods detected matching abnormal clones (r = 0.93) but the DNA index was higher than predicted by the chromosome numbers. Most of the discrepancies might be explained by tumor heterogeneity and insufficient numbers of cells available for cytogenetic analyses. In seven cases, single-cell abnormalities were found that corresponded to a flow cytometry peak. Multiclonality was present in 25% of aneuploid tumors. No association was found between metaphases in cytogenetic preparations and increased S-phase fraction of the tumors, but aneuploid tumors had a significantly higher proliferation rate. Pooling data from both methods demonstrated that the majority of our samples were aneuploid (74%).

Breast Neoplasms↗

PubMed1993

Neural Network Analysis of DNA flow cytometry histograms.

A pattern recognition system based on Neural Network Analysis, a form of artificial intelligence, was used to search DNA flow cytometry histograms for features that correlated with breast cancer patients' risk of relapse. DNA flow cytometry histograms and clinical follow-up information from 796 breast cancer patients were used to train a Neural Network to predict the clinical outcome of patients in a separate independent set of 794 patients. Median follow-up in this patient data base was short, 23 months. Neural Network Analysis resulted in a model that evaluated DNA flow cytometry histograms differently than conventional analysis, which categorizes the histograms by ploidy and S-phase fraction. Neural Network Analysis appeared to identify low risk and high risk subsets of patients as accurately as conventional analysis. Neural Network Analysis placed heavy emphasis on the region to the right of the diploid G2/M peak, where a subpopulation of nuclei with high DNA content is seen even in many histograms scored as diploid by conventional techniques. The number of nuclei in this region was found to be a powerful predictor of patient outcome, and multivariate analysis showed that the number of nuclei in this region and the S-phase fraction both were independently predictive of relapse. This pilot study suggests that conventional analysis (based on a mechanistic interpretation of regions in flow cytometry histograms) might be used in conjunction with and improved by pattern recognition systems or insights derived from them.

Breast Neoplasms↗

PubMed1980

Dual parameter flow cytometry studies in human lymphomas.

Dual parameter flow cytometry studies using Coulter volume and cell DNA content were carried out in monodisperse cell suspensions of 64 samples of human lymphoma, chronic lymphocytic leukemia, hairy cell leukemia, and benign lymphoid proliferations. Differences in mean Coulter volume among the lymphomas were due both to the intrinsic differences in mean G1 cell Coulter volume and to the presence of increased fractions of larger S and G2 cells, especially among the large B cell lymphomas. However, the relative contribution of large non-G1 cells to the overall population Coulter volume distribution was a relatively minor one; the presence of cells in S did not increase mean Coulter volume by more than 10%, even in samples with high S fractions. There was a good correlation between mean G1 cell Coulter volume and the log of the fraction of cells in S among the B cell lymphomas (r = 0.55). Evidence is presented that within individual samples, large cells proliferate more rapidly than small cells. This was seen in every case, both in the normal samples and in the lymphomas, and in the T cell lymphomas as well as in the B cell lymphomas. Aneuploidy was detected by flow cytometry in 11 cases; in 7 cases the aneuploid cell component could be analyzed separately from the diploid cell component on the basis of cell Coulter volume differences. The aneuploid components of diploid-aneuploid mixtures had higher S fractions than the diploid components in six of seven cases (0.16 +/- 0.04 [SE] vs. 0.08 +/- 0.02). These findings are considered in relation to the histopathological classification of the lymphomas, and in relation to the concept of clonal selection and clonal evolution of tumors.

Aneuploidy↗

PubMed1994

[Lymphocyte subpopulations determined by flow cytometry in chronic type B lymphatic leukemia].

BACKGROUND: For examination of surface characteristics of lymphocytes automatic analyzers of flow cytometry are used which make it possible to assess labelling with 2-3 monoclonal antibodies on one examined cell and provide thus more accurate data on the immunophenotype of proliferating cells. The objective of the present work was-using the method of flow cytometry and monoclonal antibodies-to detect surface signs on lymphocytes in the peripheral blood stream of patients with lymphatic leukaemia. METHODS AND RESULTS: In 29 patients with chronic lymphatic leukaemia type B (B-CLL) CD signs were examined, using flow cytometry and the findings were compared with those obtained in a control group of 63 healthy subjects. Evidence of CD 19+ CD5+ lymphocytes (B1 cells) in B-CLL (83.4 +/- 15% vs. 3.5 +/- 1.9% in controls) was of diagnostic importance. These CD5+ B cells are polyreactive, "memory B" lymphocytes which contrary to "conventional" lymphocytes B2 (CD5-B) do not differentiate further to plasma cells. Accumulation of B1 (CD5+ B) lymphocytes was, on the other hand, associated in patients with B-CLL with a decrease of B2 (CD5-B) lymphocytes (2.8 +/- 4.0% vs 9.0 +/- 4.2%) (p < 0.01). This finding can in case of excess of B1 (CD5+ B) lymphocytes explain the tendency of patients with B-CLL to develop autoimmune complications (e.g. AIHA); and conversely a decrease of B2 (CD5-B) lymphocytes can lead to hypogammaglobulinaemia which is also associated with B-CLL. A tendency towards autoimmunity may be also promoted by a decline of so-called inductors of suppressor cells (CD 4+ CD45RA); in patients with B-CLL among CD4 T lymphocytes 32.5 +/- 15% CD 45RA+ cells were found, as compared to 43.0 +/- 14.8% in controls (p < 0.011) and a lower ratio of true suppressor T cells (CD8+ CD11b+) in patients with B-CLL: 47.0 +/- 17.6% CD11b+ from CD8+ cells, as compared with controls 80.4 +/- 13.3 CD11b from CD 8+ cells (p < 0.01). CONCLUSIONS: The method of flow cytometry with double fluorescence thus contributes to the accurate diagnosis of B-CLL, to monitoring of the therapeutic effect but also to knowledge of the immunopathology of the disease.

Adult↗

PubMed2000

Detection of anti-HLA antibody by flow cytometry in patients with a left ventricular assist device is associated with early rejection following heart transplantation.

BACKGROUND: Patients with a left ventricular assist device (LVAD) as a bridge to heart transplantation (HT) often have elevated levels of panel reactive antibodies (PRA). The clinical significance of anti-human histocompatibility leukocyte antigen (HLA) antibodies detected by flow cytometry in PRA negative patients remains unclear. METHODS: Eighteen patients who underwent LVAD placement as a successful bridge to HT had standard anti-human globulin complement-dependent cytotoxicity and retrospective flow cytometry assays performed to detect class I anti-HLA antibodies. A positive flow result was defined as a fluorescent ratio of 23:1 versus a negative control. RESULTS: Six patients had anti-HLA antibodies detected by flow cytometry. Univariate analysis demonstrated more moderate-severe rejection episodes (ISHLT > or = IIIA) at 2 months (0.83+/-0.75 vs. 0; P=0.04) and a trend toward decreased time to first rejection (61+/-17 vs. 225+/-62 days; P=0.06) in these patients. No differences were observed in donor-recipient HLA mismatch or 1 year Kaplan-Meier survival between patients with or without anti-HLA antibodies. CONCLUSION: Despite a negative PRA, LVAD patients with class I anti-HLA antibodies detected by flow cytometry have a greater incidence of moderate-severe rejection in the first 2 months after HT. Flow cytometry may be a useful clinical tool in screening PRA negative LVAD patients before transplantation. Patients with positive anti-HLA antibody screening by flow cytometry may require more intensive immunosuppression in the early post-HT period.

Adult↗

PubMed2005

Comparison of immunocytochemistry, real-time quantitative RT-PCR and flow cytometry for detection of minimal residual disease in neuroblastoma.

A quantitative and precise measure of treatment response is warranted in neuroblastoma patients. We compared three quantitative methods often used for detection of minimal residual disease in such patients. Specificity, sensitivity and concordance of immunocytochemistry, real-time quantitative reverse transcription polymerase chain reaction (RT-PCR) and flow cytometry were compared using experimental cell suspensions (n = 8) and clinical samples (n = 126). Neuroblastoma cells were identified by immunocytochemistry and flow cytometry using anti-GD2 (14.G2a) and anti-NCAM (5.1H11) antibodies, whereas tyrosine hydroxylase mRNA was the molecular target for quantitative RT-PCR. The sensitivity using flow cytometry was 1-2 logs less than using immunocytochemistry or quantitative RT-PCR. All control samples (n = 35) tested negative by immunocytochemistry, whereas 2/34 (6%) and 1/14 (7%) were false positive by quantitative RT-PCR and flow cytometry respectively. Concordant results were obtained in 85% of patient samples (n = 116) analyzed in parallel by quantitative RT-PCR and immunocytochemistry, whereas 71% of samples analyzed by flow cytometry and immunocytochemistry were concordant (n = 35). The correlation between tumor cell levels analyzed by quantitative RT-PCR and immunocytochemistry was high (r = 0.78, p < 0.001). Quantitative RT-PCR and immunocytochemistry both reliably detected very low levels of neuroblastoma cells in clinical samples. The agreement and correlation between these methods were high. In comparison, flow cytometry was less sensitive.

Antibodies↗

PubMed1993

Applicability of flow cytometry in breast cancer.

The cumulative evidence indicates that flow cytometry can provide important information about the behavior of breast tumors. A particularly appealing aspect of this technology is the ability to determine flow cytometric parameters on fresh surgical samples, frozen biopsy specimens, and archival paraffin-embedded blocks. However, several issues regarding the standardization of tissue preparation and the cell cycle analyses must be addressed before flow cytometry can be routinely applied on a wide scale. Quality control standards are currently being discussed that will help to reduce interlaboratory variability, particularly with respect to absolute values of S-phase fraction.

Breast Neoplasms↗

PubMed1980

Deoxyribonucleic acid flow cytometry of germ cells in the investigatin of male infertility.

DNA flow cytometry was applied to fine-needle aspiration biopsy specimens from the testes of 20 oligospermic or azoospermic men under investigation for infertility. All aspirations gave sufficient material for DNA flow cytometry, and no complications occurred after aspiration. In both groups many DNA patterns deviated from those observed in control specimens. In general, abnormally small testes were associated with the greatest deviations in DNA patterns. In specimens from certain azoospermic men the DNA distributions did not deviate from those of control samples, suggesting a block in the testicular excurrent ducts. These findings also suggest that DNA flow cytometry of aspirated testicular material can be used in the classification of tubular degeneration in males. The results are obtained within a short time, are quantitative and without bias, and the aspiration causes only little discomfort to the patient. The method can therefore be recommended as a very practical means of obtaining quantitative estimates of spermatogenesis in men being evaluated for infertility.

DNA↗

PubMedDate not supplied

Flow cytometry: clinical applications in equine medicine.

The use of flow cytometry in veterinary diagnostics is becoming a valuable clinical tool with a broad range of applications. Physical characteristics of cells can be determined by the flow cytometer laser and electronics through the measurement of changes in light scatter properties. Other components and functions of cells can be defined through the application of fluorochrome dyes that have an affinity for cellular components. Traditionally, common clinical applications are immunophenotyping of cells of the hematopoietic system with fluorescent-labeled antibodies raised against specific cell surface proteins. Other approaches have been used to elucidate changes in cell function and DNA content. This review is intended to provide the reader with the fundamental uses of flow cytometry. Examples of clinical applications in equine patients include immune-mediated hemolytic anemia, immune-mediated thrombocytopenia (IMT), chronic inflammatory disease, and neoplasia.

Animals↗

PubMed2001

Validation of flow cytometry for assessment of viability and acrosomal integrity of dog spermatozoa and for evaluation of different methods of cryopreservation.

The aims of the present study were: (i) to validate the accuracy of flow cytometry for assessment of viability and acrosomal status of canine spermatozoa; and (ii) to evaluate the cryopreservation protocols currently used for dog spermatozoa using flow cytometry. Data obtained by flow cytometry analysis of fresh dog spermatozoa stained with carboxyfluorescein diacetate (CFDA) and propidium iodide, or with fluorescein isothiocyanate (FITC)-conjugated Pisum sativum agglutinin (PSA) and propidium iodide, were compared with those obtained by microscopic evaluation. The results demonstrated that flow cytometry is a precise method for evaluating the viability and acrosomal status of fresh samples of dog semen. A new triple staining procedure, using carboxy-SNARF-1, propidium iodide and FITC-PSA, was developed and was an efficient method for evaluating the following aspects of cryopreservation protocols for dog spermatozoa: (i) addition of 0.5% (v/v) Equex STM paste to a Tris-egg yolk-based extender; (ii) dilution of the semen in one or two steps; (iii) freezing semen by placing 0.5 ml straws horizontally above liquid nitrogen in a styrofoam box or lowering them vertically into a liquid nitrogen tank; (iv) thawing semen at two different rates; (v) packaging semen at different sperm concentrations; and (vi) diluting semen at different rates after thawing. The highest sperm survival and longevity was obtained when Equex was present in the semen extender, the semen dilution was performed in two steps to obtain a concentration of 2.0 x 10(8) spermatozoa ml-1, the freezing was carried out using the styrofoam box, the straws were thawed at 70 degrees C for 8 s and the semen was diluted 1:4 after thawing.

Acrosome↗

PubMed1996

Comparison of flow cytometry and western blotting to measure Hsp70.

The levels of constitutive and inducible forms of heat shock protein 70 (hsp73 and hsp72, respectively) through the cell cycle were measured in CHO cells by flow cytometry and Western blotting at various times after heating. Cells were labeled with antibody C92 (hsp72) or N27 (hsp73) and propidium iodide prior to analysis by flow cytometry. Cells were heated for 15 min at 45 degrees C, then analyzed from 3 to 36 h later. There was about a tenfold increase in hsp72 in early S phase cells beginning within 6 h after heating and these cells gradually cycled though S phase so by 36 h most of them had divided. When CHO cells were exposed to 10 microM sodium vanadate, an inhibitor of tyrosine phosphatase, for 24 h prior to heating, the induction of hsp72 in early S phase cells was almost completely inhibited. Heated cells did not express hsp73 in a cell-cycle-dependent manner. Hsp73 increased uniformly in all cells by 10 h after heating and sodium vanadate did not affect the expression. Quantitative comparisons of the relative levels of hsp72 and hsp73 measured by flow cytometry and Western blotting were in excellent agreement. Control and heated cells were labeled with Hoechst 33342 and sorted from G1, S, and G2/M phases and processed by Western blotting to verify the cell cycle dependent increase in hsp72 as measured by flow cytometry. Again there was excellent agreement between the Western blotting and flow cytometry results.

Animals↗

PubMed2004

Stage-specific activity of potential antimalarial compounds measured in vitro by flow cytometry in comparison to optical microscopy and hypoxanthine uptake.

The evaluation of new antimalarial agents using older methods of monitoring sensitivity to antimalarial drugs are laborious and poorly suited to discriminate stage-specific activity. We used flow cytometry to study the effect of established antimalarial compounds, cysteine protease inhibitors, and a quinolone against asexual stages of Plasmodium falciparum. Cultured P. falciparum parasites were treated for 48 h with different drug concentrations and the parasitemia was determined by flow cytometry methods after DNA staining with propidium iodide. P. falciparum erythrocytic life cycle stages were readily distinguished by flow cytometry. Activities of established and new antimalarial compounds measured by flow cytometry were equivalent to results obtained with microscopy and metabolite uptake assays. The antimalarial activity of all compounds was higher against P. falciparum trophozoite stages. Advantages of flow cytometry analysis over traditional assays included higher throughput for data collection, insight into the stage-specificity of antimalarial activity avoiding use of radioactive isotopes.

Animals↗

PubMed2006

In vivo micronucleus test with flow cytometry after acute and chronic exposures of rats to chemicals.

The use of flow cytometry with rat peripheral blood erythrocytes is expected to increase the sensitivity of the in vivo micronucleus test and allows assessment of the genotoxic effects at doses that may be equal or close to those relevant to human exposure. However, there was a limitation to the use of rat peripheral blood erythrocytes since the spleen selectively removes micronucleated erythrocytes from circulation. In the present study, the selective analysis by flow cytometry of young MN-PCEs (micronucleated polychromatic erythrocytes or reticulocytes) by use of anti-CD71 antibodies was intended to compensate for the splenic clearance of micronucleated erythrocytes. The young polychromatic erythrocytes have on their surface a specific marker (CD71 antigen) that decreases in density during the maturation process. To investigate the usefulness of the flow cytometric micronucleus analysis combined with anti-CD71 staining of reticulocytes several compounds were tested in acute or sub-chronic treatment regimens. Furthermore, an evaluation was conducted in comparison with the standard rat bone-marrow micronucleus test with additional compounds. The results of acute studies with intraperitoneal application of ethyl methanesulfonate (EMS) (50, 100 and 200 mg/kg) and mitomycin C (MMC) (0.5, 1 and 2 mg/kg), were comparable to data published in the literature. Sub-chronic experiments were performed with cyclophosphamide (CP) (1, 2, 4 and 8 mg/(kg day)), colchicine (6, 8 mg/(kg day)) and mitomycin C (0.1 mg/(kg day)) and showed dose- and time-dependent accumulation of MN-PCEs. Parallel analysis of micronucleus induction in peripheral blood and bone marrow performed with Novartis compounds up to the highest tested dose (5 mg/kg of compound A, 200 mg/kg of compound B and 1250 mg/kg of compound C) showed concordant results. Furthermore, we performed kinetic studies of micronucleus induction in peripheral blood samples obtained at various times after a single treatment with 10 mg/kg CP and with 6 or 8 mg/kg of colchicine. Such experiments gave important supplementary information about the time course of micronucleus induction. Our data suggest that the peripheral blood flow-cytometry micronucleus test can be used for the assessment of micronucleus induction after acute and chronic exposures of rats to chemicals.

Animals↗

PubMed1994

Correlation between immunohistochemical expression of proliferating cell nuclear antigen and flow cytometry parameters in colorectal neoplasia.

PURPOSE: Proliferating cell nuclear antigen immunohistochemical expression and flow cytometry techniques were used in this study to estimate the proliferation tendency and biologic aggressiveness in benign and malignant epithelial tumors of the colon and rectum. METHODS: Thirty-five adenomas and 60 adenocarcinomas were studied immunohistochemically concerning proliferating cell nuclear antigen positivity in tumor cell nuclei. In addition, flow cytometry techniques were used to estimate the DNA content and percentage of tumor cells in the S-phase. RESULTS: The mean proliferating cell nuclear antigen score for adenomas was 38 percent compared with a mean score of 50.4 percent for adenocarcinomas that were studied (P < 0.05). In dysplastic areas of malignantly transformed adenomas (n = 5), the highest proliferating cell nuclear antigen score (80 percent) was focally observed. Taking flow cytometry parameters into account, we found out that proliferating cell nuclear antigen can be used as an indirect indicator of the number of cells in the S-phase (SPF) but not as an independent prognostic factor. Statistical significance was found between Type III (aneuploid carcinomas) and increased proliferating cell nuclear antigen expression (proliferating cell nuclear antigen score > or = 60 percent). Furthermore, aneuploidy was especially found on cancer located on the left colon (44 percent vs. 14 percent of right colon neoplasms). Considering DNA ploidy of the above neoplasms, the aneuploid adenocarcinomas had a tendency for poorer prognosis especially if they were related to Dukes Stage C female patients. CONCLUSIONS: The comparative assessment of the above parameters might be of considerable importance in the study of the proliferation activity of any form of colorectal neoplasia.

Adenocarcinoma↗