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PubMed1997

Possibility of correcting image cytometric nuclear DNA (ploidy) measurements in tissue sections. Insights from computed corpuscle sectioning and the reference curve method.

OBJECTIVE: To describe, validate and apply a computer simulation of corpuscle sectioning to determine which assumptions are compatible with the possibility of correction. STUDY DESIGN: A computer program was written in PASCAL and run on a personal computer. RESULTS: Correction is practical in the case of spherical nuclei with homogeneous intranuclear DNA and a constant internuclear DNA concentration, even if section thickness is uneven or inaccurate. Correction is not possible in the case of marked nuclear ellipticity: mild cases are correctable, although subpopulations in a mixed-ploidy sample would not be distinguished. Correction is possible in the case of spherical (but not ellipsoidal) nuclei with a variable internuclear DNA concentration, and subpopulations in a mixed-ploidy sample would be distinguishable if sufficiently different. If the DNA is markedly concentrated in one part of the nucleus, overcorrection results. CONCLUSION: The results clarify the factors that limit correction of image cytometric measurements of ploidy in tissue sections. A new method of correcting ploidy measurements in tissue sections is usable.

Cell Nucleus↗
PubMed1996

Ultrastructural changes related to multidrug resistance in CEM cells: role of cytoplasmic vesicles in drug exclusion.

The multidrug resistance phenotype is found to be frequently associated with the overexpression of proteins which lead to a decrease of drug accumulation within human tumor cells. A 170 kDa membrane glycoprotein which is related to the overexpression of the mdr1 gene is inserted in the plasma membrane and pumps the cytotoxic drugs out of the cells. The aim of this work was to study the morphological modifications of resistant CEM/VLB 100 cells relative to their parental drug-sensitive ones and the detection of the ultrastructural localization of P-glycoprotein at the cytoplasmic level. Using a scanning electron microscope, CEM resistant cells showed wide smooth protrusions while CEM sensitive cells showed microvilli and fine folds. With transmission electron microscopy, an enhanced secretory system was observed in CEM resistant cells: both electron transparent and electron opaque vesicles were associated with the Golgi system, revealed by wheat germ agglutinin-colloidal gold labelling. These vesicles were the binding site of C 219 and MRK 16 antimembrane glycoprotein antibodies, and some of them were determined to belong to the lysosomal system after PTA staining. These vesicles may be an additional way to decrease the cellular uptake of drugs in multidrug resistant cells. Moreover, some nuclear and nucleolar modifications were also observed. These observations show that MDR has wide morphological features which concern several organelles.

ATP Binding Cassette Transporter, Subfamily B, Mem↗
PubMed1997

Variability in cytokine and cell adhesion molecule staining in arthroscopic synovial biopsies: quantification using color video image analysis.

OBJECTIVE: To investigate the variability in immunostaining for cytokines and cell adhesion molecules using multiple arthroscopically directed synovial biopsies from within a rheumatoid knee joint, quantitated by color video image analysis. METHODS: Needle arthroscopic biopsies were taken from multiple sites (4-7 sites) around a knee joint in 8 patients with rheumatoid arthritis (RA). In 5 patients, immunoperoxidase staining for the cytokines tumor necrosis factor alpha (TNF-alpha), interleukin 8 (IL-8), and IL-1beta as well as the IL-1 receptor antagonist protein (IL-1ra) was performed. In 3 patients, immunoperoxidase staining for the cell adhesion molecules E-selectin (CD62E), P-selectin (CD62P), intercellular adhesion molecule 1 (ICAM-1, CD54), and platelet endothelial cell adhesion molecule (PECAM, CD31) was performed. Immunostaining was quantified using color video image analysis. RESULTS: The overall probability of paired biopsies from the same RA knee joint being significantly different from each other due to sampling variation was at most 22% for cytokine staining (usually less than 10%). There were no significant differences between intrabiopsy and interbiopsy variability for cell adhesion molecule staining of the sublining and vessels. CONCLUSION: The variability in cytokine and cell adhesion molecule staining within any single biopsy usually reflects the variability between biopsies taken from different sites in the same rheumatoid joint when the immunostaining is quantified using color video image analysis. Therefore, only a small number of synovial biopsies are required to accurately determine the cytokine and cell adhesion molecule expression in a single joint.

Aged↗
PubMed1998

Cholesteatoma: a molecular and cellular puzzle.

HYPOTHESIS: There are at least three possible molecular models of cholesteatoma pathogenesis. Cholesteatoma may arise as a result of 1) the induction of a preneoplastic or neoplastic transformation event; 2) a defective wound-healing process; and/or 3) a pathologic collision of the host inflammatory response, normal middle ear epithelium, and a bacterial infection. BACKGROUND: There have been a number of speculations concerning the factors that foster the development of cholesteatoma. Before resolving the molecular basis for the pathogenesis of cholesteatomas, it is important to present and test plausible models that could explain how a cholesteatoma becomes invasive, migratory, hyperproliferative, aggressive, and recidivistic. METHODS: The authors evaluated by various techniques (e.g., immunohistochemistry, flow cytometry, and image analysis) a large number of cholesteatomas of all types (e.g., primary and secondary acquired, recurrent, and congenital) and a range of normal tissues (tympanic membrane, canal wall skin, and postauricular skin) for the expression of various proteins (e.g., p53, ectopeptidases, tryptase) and for the presence of DNA aneuploidy. RESULTS AND CONCLUSIONS: The authors' published and unpublished studies to date support several suppositions concerning the pathology of cholesteatomas. First, cholesteatoma epithelium behaves more like a wound-healing process than a neoplasm. The available evidence to date does not indicate that cholesteatomas have inherent genetic instability, a critical feature of all malignant lesions. Second, the induction of hyperproliferative cells in all layers of the cholesteatoma epidermis implicates a potential idiopathic response to both internal events as well as external stimuli in the form of cytokines released by infiltrating inflammatory cells. Third, the presence of bacteria may provide a critical link between the cholesteatoma and the host, which prevents the cholesteatoma epithelium from terminating specific differentiation programs and returning to a quiescent state in which it becomes minimally proliferative, nonmigratory, and noninvasive. Fourth, none of our data suggest that there are any obvious molecular or cellular differences among the various types of cholesteatomas (e.g., primary and secondary acquired, recidivistic, and congenital). Continued research should delineate the precise molecular and cellular dysfunction involved in the pathogenesis of cholesteatomas and how this knowledge can be useful in the clinical management of cholesteatomas.

Aminopeptidases↗
PubMed1998

The receptor for complement anaphylatoxin C3a is expressed by myeloid cells and nonmyeloid cells in inflamed human central nervous system: analysis in multiple sclerosis and bacterial meningitis.

The complement anaphylatoxins C5a and C3a are released at the inflammatory site, where they contribute to the recruitment and activation of leukocytes and the activation of resident cells. The distribution of the receptor for C5a (C5aR) has been well studied; however, the receptor for C3a (C3aR) has only recently been cloned, and its distribution is uncharacterized. Using a specific affinity-purified anti-C3aR peptide Ab and oligonucleotides for reverse transcriptase-PCR analysis, C3aR expression was characterized in vitro on myeloid and nonmyeloid cells and in vivo in the brain. C3aR was expressed by adult astrocytes, astrocyte cell lines, monocyte lines THP1 and U937, neutrophils, and monocytes, but not by K562 or Ramos. C3aR staining was confirmed by flow cytometry, confocal imaging, and electron microscopy analysis. A 65-kDa protein was immunoprecipitated by the anti-C3aR from astrocyte and monocyte cell lysates. Our results at the protein level were confirmed at the mRNA level. Using reverse transcriptase-PCR, Southern blot, and sequencing we found that C3aR mRNA was expressed by fetal astrocytes, astrocyte cell lines, and THP1, but not by K562 or Ramos. The astrocyte C3aR cDNA was identical with the reported C3aR cDNA. C3aR expression was not detected in normal brain sections. However, a strong C3aR staining was evident in areas of inflammation in multiple sclerosis and bacterial meningitis. In meningitis, C3aR was abundantly expressed by reactive astrocytes, microglia, and infiltrating cells (macrophages and neutrophils). In multiple sclerosis, infiltrating lymphocytes did not express C3aR, but a strong staining was detected on smooth muscle cells (pericytes) surrounding blood vessels.

Adult↗
PubMed1998

In situ subcellular localization of epitope-tagged human and rabbit carboxylesterases.

Carboxylesterases are a ubiquitous class of enzymes thought to be involved in xenobiotic metabolism and detoxification. Primary amino acid sequence data suggest that these proteins localize to the endoplasmic reticulum. However, since this family of proteins is highly homologous, the generation of specific reagents to monitor expression and subcellular localization has been unsuccessful. To accomplish in situ detection of a human alveolar macrophage carboxylesterase and a rabbit liver carboxylesterase, we constructed plasmids that expressed recombinant proteins containing an 11 amino acid influenza hemagglutinin tag near the C-terminus. These proteins retained carboxylesterase activity as determined by the conversion of o-nitrophenol acetate to o-nitrophenol. Following transfection of plasmids encoding these proteins into mammalian cells, cells were analyzed by both fluorescence and electron microscopy. The tagged enzymes were localized to the endoplasmic reticulum of both Cos7 monkey kidney cells and Rh30 human rhabdomyosarcoma cells. No tagged protein was detectable in the culture media. Hence, epitope tagging allowed the analysis of expression and localization of specific carboxylesterases. The methods described in this paper are, therefore, applicable to any protein, including those that are highly homologous to other candidate molecules.

Amino Acid Sequence↗
PubMed1998

Association of prognosis in surgically treated lung cancer patients with cytometric, histometric and ligand histochemical properties: with an emphasis on structural entropy.

OBJECTIVE: To explore new tumor features for refined category formation that permits the tailoring of individualized treatment schemes in lung cancer. STUDY DESIGN: Survival data on patients from six independent studies on cases with surgically treated lung cancer, primary lung carcinoids or metastasizing breast carcinoma (including data on primary breast carcinoma) were analyzed by nonhierarchic multivariant discriminant analysis with respect to a set of cytometric/histometric and immunohistochemical/ligand histochemical parameters. The number of stem lines, S-phase-related tumor cell fraction and the extent of structural entropy and its current were measured. In addition, the expression of binding capacities for histo-blood group trisacharides, galectins, the alpha/beta-interferon antagonist sarcolectin, the lymphokine macrophage migration inhibitory factor and a monoclonal antibody to the Le(y) epitope was monitored for insight into aspects of immunologic and biologic behavior. RESULTS: In all studies, a correlation between tumor parameters, according to TNM stage and survival, was seen. In order to refine this category formation, at least certain selected features should provide an even more stringent association than TNM stages. Indeed, statistical correlation of the cytometric and histometric parameters as well as the expression of receptors for the two histo-blood group trisaccharides, ligands for the galectins (CL-16, CL-14) and macrophage migration inhibitory factor was stronger than that of TNM stage. A large amount of the current of structural entropy was especially highly significantly associated with poor survival. This observation could be verified in each of the different studies. CONCLUSION: The obtained data strongly support the notion that thermodynamic evaluation of tumor growth focusing on the "entropy distance" of the tumor from its environment is a promising perspective warranting extended studies. Additionally, glycohistochemical features, including binding capacities for histo-blood group trisaccharides, have the potential to aid in establishment of a biologic marker set for tumor staging.

Biomarkers, Tumor↗
PubMed1998

The role of biopsy pathology, quantitative nuclear morphometry, and biomarkers in the preoperative prediction of prostate cancer staging and prognosis.

The focus of this review is to survey pretreatment biopsy and patient-derived information applicable to predicting pathological stage and prognosis of men with a diagnosis of prostate cancer. Various sources of clinical and pathological information that may contribute to building decision support tools (DSTs) for application by the urologist to manage prostate cancer patients are presented. These DSTs use serum biomarkers and objective, well-established, pathology information extracted by experienced pathologists from needle-core tissue samples that describe tumor size, grade, and location. Other valuable data can be derived from the biopsy tissue, such as computer-assisted image cytometry-derived DNA ploidy and nuclear morphometry informatics, as well as select tissue biomarker results that may provide supplemental prognostic information. Also discussed are the technical and clinical limitations of these DSTs with respect to the prediction accuracy. A commercially available pretreatment prediction algorithm (UroScore, Oklahoma City, OK) was applied to predict the disease organ confinement status of the prostate cancer test case. Finally, the authors present existing and future applications of computer-derived computational solutions for incorporating all patient history, clinical laboratory, and pathology information into algorithms that can generate patient-specific predictive probability estimates of stage, recurrence, and progression.

Biomarkers, Tumor↗
PubMedDate not supplied

Prognostic relevance of histomorphological parameters and DNA content and their therapeutic consequences in esophageal carcinoma: a multivariate approach.

BACKGROUND/AIMS: Despite recent advances in surgical and multidisciplinary treatment, the prognosis for patients with esophageal carcinoma remains poor. The low prognostic accuracy of even surgical pathological TNM staging suggests that additional parameters would be useful in determining the prognosis. METHODOLOGY: We undertook a retrospective analysis of 115 patients who had undergone transhiatal and transthoracic esophageal resection due to squamous cell carcinoma of the esophagus. In addition to TNM classification and the usual morphological criteria, a quantitative DNS analysis using Image DNA cytometry was performed. At the time of DNA analysis, histomorphological parameters and survival time was not known. RESULTS: The main prognostic parameter was the curativity (R classification) of the operation. Using only patients who had had R0 resection, a multivariate analysis was performed. Parameters included: patient age; preoperative ASA classification; tumor localization; pN category; number of intra-thoracal lymph nodes removed; pT category; pM category; grading; lymphangiosis; ploidy; operative procedure, and the development of postoperative complications identifying the ploidy, and the depth of tumor infiltration of prognostically independent factors. CONCLUSIONS: In the case of a diploid or tetraploid DNA content, tumor resection is recommended even in the case of lymph node metastasis at the truncus coeliacus. Patients with a diploid or tetraploid tumor without distant metastasis and tumor stage pT1-pT3 may, after curative (RO) transthoracic resection with two-field lymph node dissection, have an advantage over patients having a transmediastinal procedure in terms of long-term follow up. In cases of aneuploid DNA content, tumor resection shows no advantage over palliative non-operative procedures. Preoperative radio- or chemotherapy may improve the prognosis of these patients.

Adolescent↗
PubMedDate not supplied

[Thyroid glands as an object of morphometric study].

Standardization of the morphometric studies of the thyroid is performed. The following table for a semiquantitative analysis of thyroid morphofunctional state is given: 1) epithelial components with characteristics of main cell types and their proliferative activity; 2) stromal components; 3) some pathologic changes (cyst, hemorrhages, etc.). A quantitative study of the thyroid includes: I) on the organ (macroscopic level)--thyroid volume, weight, length, width, height; II) light microscopy: point counting, stereological methods. Main parameters: 1) epithelial components (18 indices are suggested); 2) proliferative activity (7 indices: nuclear diameter and surface, mitotic activity, etc.); 3) desquamation (mean number of cells per follicle, relative volume of the desquamated epithelium); 4) colloid characteristics; 5) stromal characteristics; 6) vascular characteristics; III) electron microscopy. The first stage of the morphometric study is a description, then semiquantitative method supported by a deep morphometric analysis with the use of statistics.

Animals↗
PubMed1998

Nuclear chromatin texture in prostatic lesions. I. PIN and adenocarcinoma.

OBJECTIVE: To document changes in the chromatin pattern in secretory cell nuclei from prostates with prostatic intraepithelial neoplasia (PIN) or adenocarcinoma. STUDY DESIGN: High-resolution images of nuclei were recorded, and a set of features descriptive of the chromatin texture and spatial distribution was computed. From this data set, features undergoing a monotonic trend of progression were selected and plotted to reveal trends in lesion progression. RESULTS: The nuclear chromatin in secretory cells in prostates with either PIN or malignant adenocarcinoma undergoes distinct and statistically significant changes in its texture and spatial distribution. Two trends of progressive change were observed. First, the values of a number of features descriptive of the clumpiness of the chromatin increase from values found in normal prostates to those recorded for nuclei from low grade to high grade PIN lesions. The second trend is a decrease in the values of the same features from those found in nuclei from high grade PIN still facing an intact basal cell layer to those no longer facing such a layer. This may be the first detectable step in progression towards development of a malignant lesion. There is a further decrease in nuclei in glands immediately adjacent to adenocarcinoma and in malignant lesions themselves. CONCLUSION: The described changes may lend themselves to the monitoring of lesion progression or of response to treatment or to chemopreventive intervention.

Adenocarcinoma↗
PubMed1998

Nuclear chromatin texture in prostatic lesions. II. PIN and malignancy associated changes.

OBJECTIVE: The objective of this study was to identify and document prostatic intraepithelial neoplasia (PIN) and malignancy associated changes in secretory cell nuclei from visually normal appearing tissue regions of prostates harboring PIN or adenocarcinoma. STUDY DESIGN: High-resolution digitized images of nuclei were recorded in histologically normal appearing tissue regions at defined distances from the margin of PIN or malignant lesions. Features descriptive of nuclear chromatin texture were computed and used to derive a discriminant function score for each nucleus. RESULTS: Secretory cell nuclei in prostates harboring either PIN or adenocarcinoma were shown to have statistically significantly different chromatin texture from secretory cell nuclei recorded in prostates free from any such lesion. The expression of PIN or malignancy associated changes was documented for distances up to 10 mm from the margin of a lesion. CONCLUSION: The finding of characteristic changes in nuclear chromatin texture of nuclei from histologically normal appearing tissue in prostates with PIN or adenocarcinoma offers the potential for higher sensitivity of detection of such lesions and for earlier detection of changes potentially preceding the development of clinically significant disease.

Cell Nucleus↗
PubMed1998

Chromatin texture signatures in nuclei from prostate lesions.

OBJECTIVE: To characterize nuclei from prostatic lesions in a highly specific manner by developing a nuclear chromatin texture signature and to characterize lesions by means of their composition of nuclei with diverse degrees of deviation from normal. STUDY DESIGN: High-resolution digitized imagery of nuclei from normal prostates, from prostatic neoplastic lesions of low and high grade and from histologically normal appearing regions of prostates with low and high grade prostatic intraepithelial neoplasia (PIN) lesions were recorded. A set of 65 features descriptive of the spatial and statistical distribution of nuclear chromatin was computed for each nucleus. These features were arranged and processed to form a distinctive signature. A distance metric from "normal" was defined and computed for each nucleus. RESULTS: Profiles of feature values can, after suitable scaling, be presented as distinctive feature value signatures. For many practical applications, profiles based on a standardized distance from normal nuclei may be more useful. Such profiles allow the derivation of a progression curve, showing increasing distances for diagnostic groups with increasing lesion progression up to high grade PIN lesions. Within each diagnostic group different cases show distinctive distributions of nuclei with differing degrees of deviation from normal, allowing the derivation of a lesion signature. CONCLUSION: Nuclear chromatin texture signatures may be of value for the characterization of both nuclei and lesions. They are based on a more comprehensive use of information offered by the nuclear chromatin pattern than that included in classification methods. While these signatures offer a more specific characterization of a clinical sample, they also are subject to more variability within a diagnostic category. This may not be due to randomness but may reflect some actual differences between lesions.

Adenocarcinoma↗
PubMed1998

Automated histometry in quantitative prostate pathology.

OBJECTIVE: To review progress on the development of machine vision and image understanding in prostate tissue histology and to discuss the problems and opportunities afforded to pathology through the use of these techniques. STUDY DESIGN: A variety of concepts in machine vision are explored, and methodologies are described that have been developed to deal with the complexities of histologic imagery. The theory of human vision and its impact on machine vision are discussed. Software has been specifically developed for the analysis of prostate histology, allowing accurate gland segmentation, basal cell identification and measurement of vascularization within lesions. RESULTS: Image interpretation can be achieved using knowledge-based image analysis and the application of local object-oriented processing. This successfully allows an automated quantitative analysis of histologic morphology in the diagnosis of prostate intraepithelial neoplasia and invasive prostatic cancer. The use of low-power image scanning, based on textural or n-gram mapping, permits the development of fully automated devices for the rapid detection of tissue abnormalities. High-power, knowledge-guided scene segmentation can be carried out for the quantitative analysis of cellular features and the objective grading of the lesion. CONCLUSION: Automated tissue section scanning and image interpretation is now possible and holds much promise in prostate pathology and other diagnostically demanding areas. Issues of standardization still need to be addressed, but the development of such systems will undoubtedly enhance our diagnostic capabilities through the automation of time-consuming procedures and the quantitative evaluation of disease processes.

Humans↗
PubMed2006

Asynchronous progression through the lytic cascade and variations in intracellular viral loads revealed by high-throughput single-cell analysis of Kaposi's sarcoma-associated herpesvirus infection.

Kaposi's sarcoma-associated herpesvirus (KSHV or human herpesvirus-8) is frequently tumorigenic in immunocompromised patients. The average intracellular viral copy number within infected cells, however, varies markedly by tumor type. Since the KSHV-encoded latency-associated nuclear antigen (LANA) tethers viral episomes to host heterochromatin and displays a punctate pattern by fluorescence microscopy, we investigated whether accurate quantification of individual LANA dots is predictive of intracellular viral genome load. Using a novel technology that integrates single-cell imaging with flow cytometry, we found that both the number and the summed immunofluorescence of individual LANA dots are directly proportional to the amount of intracellular viral DNA. Moreover, combining viral (immediate early lytic replication and transcription activator [RTA] and late lytic K8.1) and cellular (syndecan-1) staining with image-based flow cytometry, we were also able to rapidly and simultaneously distinguish among cells supporting latent, immediate early lytic, early lytic, late lytic, and a potential fourth "delayed late" category of lytic replication. Applying image-based flow cytometry to KSHV culture models, we found that de novo infection results in highly varied levels of intracellular viral load and that lytic induction of latently infected cells likewise leads to a heterogeneous population at various stages of reactivation. These findings additionally underscore the potential advantages of studying KSHV biology with high-throughput analysis of individual cells.

Antigens, Viral↗
PubMed1994

Computerized image analysis and flow cytometric evaluation of ovarian borderline tumors: a study of 24 cases.

DNA content evaluation in the study of ovarian borderline tumors has been shown to be a useful adjunct to histopathologic diagnosis. This study compares flow cytometry and computerized image analysis (CIA) in evaluating the DNA content of these lesions. Twenty-four cases of ovarian borderline tumors (15 serous, 9 mucinous) were studied utilizing formalin-fixed paraffin-embedded tissue. Flow cytometry of cell suspensions and CIA of cell suspensions and paraffin sections were compared in the evaluation of DNA content. Twenty-three tumors (96%) were diploid and only 1 (4%) was aneuploid. There was 100% correlation between flow cytometry and CIA of cell suspensions. Image analysis of Feulgen-stained paraffin sections was found to be unreliable when compared with the use of cell suspensions in the evaluation of DNA content. Clinical follow-up in 17 patients showed recurrent disease in two patients with diploid tumors and no evidence of disease in the patient with an aneuploid tumor. The reported rate of aneuploidy in borderline tumors varies depending on the diagnostic method employed in evaluating DNA content. This study shows good correlation between flow cytometry and image analysis of cell suspensions and supports the low rate of aneuploidy found in these lesions by other authors utilizing these methods. The majority of borderline ovarian tumors have been shown to have a diploid DNA content; however, the finding of aneuploidy in histologically classified borderline tumors warrants close clinical follow-up since aneuploidy in these tumors has been associated with an adverse prognosis.

Aneuploidy↗
PubMed2006

Quantification of cell size distribution as applied to the growth of Corynebacterium glutamicum.

It is known that the cell size is related to the physiological state of a cell. Therefore, cell size distribution directly reflects the average physiological properties of the cell culture. Cell size distribution can be enumerated by image analysis, flow cytometry and coulter counter. In this study, image analysis was used to characterize the cell size distribution during the growth of Corynebacterium glutamicum and was further analyzed by a distribution function. The parameters of the distribution function indicate the mean value and spread of the distribution. Analysis demonstrated that the maximum specific growth rate was higher (0.67h(-1)) for the growth obtained through serial dilution of seed as compared to growth from a normal seed culture (0.53h(-1)). This was due to a greater percentage of the cell population being in the state of division for the growth through serial dilution in the mid-log phase. The measurement of the cell size distribution demonstrated that the average cell size decreased during the course of growth. The distribution function was also used to enumerate the average specific growth rate of both the conditions of the culture. The demonstrated methodology can be used to predict an average growth property of a cell culture.

Bacteriological Techniques↗
PubMed2006

Prognostic value of static cytometry in transitional cell carcinoma of the bladder: recurrence rate and survival in a group of patients at 10 years follow-up.

Many studies have indicated that nuclear DNA content evaluation can be used to predict biological behavior of transitional cell carcinoma (TCC) of the bladder. Some authors also indicated that static cytometry is more useful in DNA content analysis than flow cytometry. The aim of the present study was to evaluate the prognostic significance of DNA ploidy in TCC of the bladder, performed by using static cytometry with an image analyzer, and monitoring patients at 10 years follow-up. Thirty-one consecutive patients underwent transurethral or open surgery for bladder tumors, and neoplastic tissue samples taken from each patient were imprinted on glass slides and sent for histopathological and DNA content evaluation. DNA ploidy evaluation was performed using a CAS 200 image analyzer. Nuclear DNA content evaluation was compared to patient follow-up on recurrence, progression or survival performed 10 years after surgery. Pathological evaluation demonstrated the presence of superficial TCC in 23 patients, while 8 had an invasive bladder tumor. Twenty-nine tumor samples were adequate for DNA content measurement, with 13 showing diploid DNA content and 16 with aneuploid DNA content. At 10 years follow-up, all patients with aneuploid DNA content demonstrated a lower survival time (p=0.049) and higher recurrence rate (p=0.0346). A log-rank test demonstrated that stage, grade and nuclear DNA content are the most useful prognostic parameters for predicting the biological behavior of TCC of the bladder. These results confirm that static cytometry is a good and reliable method to evaluate DNA tumor content and considered a useful prognostic parameter for predicting recurrence rate, disease progression or survival in patients affected by bladder tumors.

Adult↗