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PubMed2004

Molecular cytogentic profiles of hepatitis C infection in patients at Sharkia Governorate, Egypt.

It has become apparent that hepatitis C virus (HCV) infection is a major risk factor for the development of hepatocellular carcinoma (HCC) worldwide. The precise mechanism by which HCV causes HCC is not known. Unlike the hepatitis B virus (HBV), HCV is not a DNA virus and does not become integrated within the genome of hepatocytes. It is more likely that HCC occurs against a background of inflammation and regeneration, associated with liver injury due to chronic hepatitis. In this study, 40 of paraffin blocks liver tissues from HCV-PCR positive patients (HBV seronegative) were examined using DNA image cytometry to evaluate its role in diagnosing HCC associated with HCV infection. Fluorescent in situ hybridization (FISH) technique using LSIZNF 217 chromosome 20q 13.2 probe was applied as well. The results showed high percentage of S-phase fraction in cases of G2S2 and G3S3 with DNA diploidy. Only two cases of G3S3 showed DNA aneuploidy with severe amplification of chromosome 20q 13.2. Consequently, DNA imaging cytometry is a good approach in differentiating dysplasia from well-differentiated HCC on top of HCV infection. In conclusion HCV has an acquired role in development of HCC through amplification of the aggressive tumor behavior oncogene LSIZNF 217 at chromosome 20q 13.2.

Adult↗
PubMed2002

Diagnostic approach of effusion cytology using computerized image analysis.

The objective of this study is to investigate whether image cytometry is a sensitive and specific method for the differential diagnosis of equivocal cells in routine cytology of effusion smears. One hundred four effusion smears were studied from routine cytologic material. Cytologically 56 (53.8%) of the smears were classified as malignant, 26 (24%) as suspicious and 22 (21.1%) as benign. Two morphometric variables (nuclear major axis length and nuclear area) of the nuclei were measured by an image analysis system. Higher values for the area were found for malignant rather than benign and suspicious cells (p < 0.0005 and p < 0.005 respectively). The same result was extracted for the nuclear major axis length values (p < 0.0005 and p < 0.0005 respectively). Values of nuclear major axis length and nuclear area didn't differ significantly between benign and suspicious cells (p = 0.071 and p = 0.066 respectively). The results show that the range of the values for suspicious cells is closer to the range of the benign cells. Cytomorphometry of the effusion smear cells may provide important information for the differentiation of atypical mesothelial cells from malignant adenocarcinoma cells.

Adenocarcinoma↗
PubMedDate not supplied

DNA--cytometric studies on xenografts of squamous cell carcinomas of the head and neck.

The primary tumors of four human head and neck carcinomas and their xenografts in nude mice were analysed with respect to their histological features and DNA-ploidy level using DNA-flow and DNA-image cytometry. The histological characteristics of the xenografts and the DNA-index did not differ from those of the parent tumors, whereas the growth rate remained unchanged during serial passaging. DNA-flow cytometric analysis revealed the presence of both aneuploid and euploid cells in both parent tumors and their xenografts. During passaging the proportion of aneuploid cells increased. Analysis of the different cell populations by DNA-image cytometry revealed the presence of aneuploid tumor cells and euploid host cells. This study demonstrates that the characteristics of the original tumors in terms of histological features and DNA-ploidy are retained after xenografting.

Animals↗
PubMed1994

Diagnostic and prognostic value of DNA ploidy and cell nuclearity in ultrasound-guided liver biopsies.

BACKGROUND: Focal nodule lesions in patients with cirrhotic livers may be visualized by using imaging techniques; however, the diagnostic and prognostic judgment of biopsies from borderline lesions may be difficult using conventional histologic criteria. METHODS: The diagnostic and prognostic value of DNA ploidy analysis determined by image cytometry of Feulgen-stained isolated hepatocytes was investigated in ultrasound-guided biopsies from 50 nodular lesions found in patients with cirrhotic livers (39 hepatocellular carcinomas [HCCs] and 11 macroregenerative nodules) and from 10 patients with livers affected by viral chronic hepatitis. Of the 11 macroregenerative nodules, 7 presented a subsequent neoplastic behavior. Specimens from the morphologically normal livers of five patients who underwent liver surgery served as control tissues. Image cytometry was performed on Feulgen-stained cytologic preparations, obtained by enzymatic digestion of formalin fixed biopsies. The DNA ploidy of the main stem line and the distribution of mononucleated and binucleated hepatocytes (nuclearity) were compared using histologic diagnosis, Edmondson's grade, tumor size, and patient follow-up. RESULTS: The main stem line was peridiploid in all benign specimens and in 31 clinically confirmed HCCs, peritetraploid in 11 HCCs, perioctaploid in 1 HCC, and aneuploid in 3 HCCs. The fraction of mononucleated polyploid hepatocytes was found to be the best diagnostic parameter in euploid HCCs and was significantly correlated with the Edmondson grade and the nodular size. Survival information was available for 43 patients, with a median observation period of 350 days. A DNA ploidy value of the main stem line greater than 3c was an important determinant of survival as a single parameter and in association with histologic grade and greatest dimension of tumor. CONCLUSIONS: This study suggests that the ploidy distribution analysis of mononucleated and binucleated hepatocytes can provide valuable information for making correct diagnoses and for predicting survival outcome for patients with HCCs.

Biopsy↗
PubMed1995

Quantitative microscopy and tumour cell proliferation.

The number of cell probes is rapidly increasing. During the last few years it has become possible to label total nuclear DNA (Feulgen), synthesized DNA (BrdU), specific A/T versus G/C rich DNA, cell cycle proteins (PCNA, Ki67), hormonal receptors (ER, PR, EGF), cdc genes and gene primary transcripts, etc. Taking advantage of this roster of cellular probes to assess cell kinetics in normal and malignant tissues implies not only quantitating their amount per cell but also analysing their intra-cellular respective distribution and inter-cellular variation. Image cytometry is the tool of choice for this purpose provided the quantitative results are properly interpreted. Confusions are often made between the respective meaning of i) reduced cell cycle speed (cell cycle duration), ii) increased proportion of proliferative cells (growth fraction), and iii) fraction of cells in S phase (SPF) and mitotic index (proportion of mitoses) which all tune the cell proliferative activity. The contribution of these biological cell population features to tumour growth is illustrated. Examples based on image cytometry are presented to define the individual tumour cell proliferation profile and tumour heterogeneity for proliferation profiles. It will be finely demonstrated how quantitative microscopy can turn the 'conventional static histological picture' of a tumour into a 'functional picture' that might support decision for therapeutic strategies.

Adenocarcinoma↗
PubMed2006

The flow cytometric analysis of premalignant and malignant lesions in head and neck squamous cell carcinoma.

We attempted to identify the molecular mechanisms involved in Head and Neck Squamous Cell Carcinoma (HNSCC) pathogenesis by measuring the nuclear DNA content (ploidy) in premalignant (potentially malignant) and malignant patients as compared to normal controls, and to determine whether DNA ploidy could be used to predict the clinical outcome. From March 2001 to December 2003, the analysis was carried out in a set of 41 patients with premalignant lesions and 79 suffering from squamous cell carcinoma of laryngeal, oesophageal, nasopharyngeal, nasal and oral lesions and 50 controls. Representative samples were taken by punch biopsy and processed using standard formol-paraffin technique for histopathological examination. Fifty micrometer thick sections of paraffin-embedded tissues were analyzed to detect the DNA content by image cytometry. Of the potentially malignant patients, 46% had diploid lesions, 37% had tetraploid lesions and 17% had aneuploid lesions. While of the patients with cancer, 90% had aneuploid lesions, 10% had diploid lesions and none had tetraploid lesions. DNA diploidy tended to occur earlier in the progression from premalignant to malignant lesions and this helps us early detection of HNSCC by DNA from lesions in high risk groups and examination of its ploidy. Knowledge of tumor cell ploidy by DNA image cytometry may facilitate the evaluation of malignant and premalignant lesions in HNSCC. The present findings are promising to supplement clinical and histopathological parameters in evaluating prognosis and to demonstrate methods that are readily applicable for routine diagnostic work.

Carcinoma, Squamous Cell↗
PubMed1999

Phase I trial of sequential topotecan followed by etoposide in adults with myeloid leukemia: a National Cancer Institute of Canada Clinical Trials Group Study.

Prolonged exposure to a topoisomerase I inhibitor may increase expression of topoisomerase II, making cells more susceptible inhibitors of that enzyme. This study was undertaken to establish the maximum tolerated dose (MTD) of a topotecan/topoisomerase II inhibitor sequential combination that may be active in acute leukemia, and to evaluate the effects of in vivo exposure to topotecan on topoisomerase II levels in leukemic blast cells as measured by image cytometry. Patients who were eligible for this phase I study had relapsed or refractory acute myeloid leukemia (< or = 2 prior regimens) or CML blast crisis (0 or 1 prior regimen). Topotecan was given as a 5 day continuous i.v. infusion and was to be escalated through three levels (1.5, 1.75 and 2.0 mg/m2 day), followed by etoposide at two dose levels (100 and 150 mg/m2) i.v. bolus days 6, 7 and 8. Topoisomerase IIalpha levels in leukemic blasts from bone marrow were measured by image cytometry prior to starting treatment, on day 5 of topotecan infusion and on day 28; and daily during topotecan in peripheral blood blasts. Dose-limiting toxicity was seen in two of six patients at the first dose level (topotecan 1.5 mg/m2/day, etoposide 100 mg/m2/day; > or = grade 3 mucositis in both cases). This cohort was expanded to 10 patients; no further non-hematologic dose-limiting toxicity was observed, but given the extent of toxicity seen, further dose escalation was judged not to be feasible. Topo IIalpha levels increased in peripheral blood blasts during the first 72 h of topotecan infusion and returned to near baseline by day 5, whereas levels appeared to decrease in bone marrow blasts by day 5 compared to pretreatment. One complete hematologic and cytogenetic remission in a patient with CML blast crisis was observed in the 10 patients evaluable for response. The sequential administration of topotecan 1.5 mg/m2/day continuous infusion for 5 days followed by etoposide 100 mg/m2/day x 3 is the recommended phase II dose for this schedule. Topotecan increases topo IIalpha expression in vivo in leukemia cells, but levels of the enzyme are cell cycle dependent. Pharmacodynamic evaluation of the sequential or combination administration of novel antileukemic agents may help improve treatment strategies in acute leukemia.

Acute Disease↗
PubMed1998

DNA quantification as a prognostic factor in gastric adenocarcinoma.

OBJECTIVE: To determine if DNA quantification, studied in cytologic samples obtained by fiberendoscopy, has predictive value in gastric adenocarcinoma. The survival times of patients in whom the tumor was the cause of death were considered variables of interest. STUDY DESIGN: Twenty-nine patients with gastric cancer, diagnosed by cytology, endoscopy and microbiopsy, were selected. The study was done over more than 10 years. Smears were stained with progressive hematoxylin and processed by computer for DNA evaluation by image cytometry. RESULTS: Four different types of histograms that directly relate to tumoral malignancy were obtained. These histograms were characterized by the value of entropy. We established four grades of aggressiveness. Then we obtained two large groups: high and low grade of malignancy. We studied the survival times in both groups and constructed a Kaplan-Meier survival curve for the high grade of malignancy group. We confirmed the results statistically and found that there was a significant relationship, with P < .05. CONCLUSION: The use of DNA quantification by image cytometry is strongly advised in daily surgery as a prognostic indicator of survival time in gastric adenocarcinoma patients.

Adenocarcinoma↗
PubMed1988

Image cytometric classification of premalignant breast disease in fine needle aspirates.

Image cytometry for the classification of fine needle aspirate (FNA) biopsies was evaluated in samples from 39 women. Eighteen of them had benign lesions, seven had premalignant lesions, nine had carcinoma in situ, and five had carcinoma. The term, premalignant, here refers to lesions with an increased risk of developing into breast cancer (atypical hyperplasia and, to a lesser extent, moderate or florid hyperplasia). The classifications by cytometry were compared with the microscopic diagnoses of the same FNA samples and of tissue from a subsequent surgical biopsy of the same area. One slide from each breast FNA sample was restained in Azure-A Feulgen. Breast epithelial cells were measured using a texture analysis program on the Leitz TAS-plus. The mean, standard deviation (SD), and interquartile range were calculated for each of 12 nuclear parameters from 200 cells per slide. A discriminant analysis was used to develop a statistical model for classifying individual samples. Six of seven atypical proliferative lesions (atypical hyperplasia and moderate hyperplasia) were identified by image cytometry, but were unrecognized by conventional microscopic examination.

Biopsy↗
PubMed1997

[Pathological features of hepatocellular carcinoma carrying hepatitis C virus antigen].

OBJECTIVE: To analyze the pathological features of hepatocellular carcinomas (HCCs) carrying different hepatitis virus antigens histopathologically and systematically. METHODS: PAP and ABC kits were used in the immunohistochemical study, and CAS-200 System was applied in the image cytometry. RESULTS: As compared with HCCs carrying HBV marker(s), the HCCs carrying HCV marker showed more cases of clear cell type (7/9 vs 4/33), better differentiation of cancers, less necrosis of hepatocytes, milder lymphocyte infiltration in the hepatic sinuses or periportal areas (P < 0.01), higher incidence of bile ductule damages, and bore a close relation to the formation of lymphoid follicle in the surrounding tissues (P < 0.05). These patients were elder, with lower grade of symptoms and better prognosis after operation. CONCLUSION: HCCs carrying only HCAg have different pathological features and clinical characteristics from which carrying HBV marker(s). The results of image cytometry are in accordance with the biological behaviour of HCC.

Adult↗
PubMed1992

Supportive role of image analysis and DNA ploidy pattern in the diagnosis of thyroid tumors.

To evaluate the supportive role of image cytometry and DNA ploidy pattern in the diagnosis of thyroid tumors, a preliminary study was performed on fine needle aspirates of 30 cases. Of these, 10 cases each were of colloid goiter, follicular neoplasm, and papillary carcinoma. The nuclear area and DNA value of 50 cells in each case were measured. The mean nuclear area in colloid goiter (69.50 + 12.62 sq.microns) was significantly lower than the mean nuclear area in a follicular neoplasm (88.71 + 15.51 sq.microns) (P less than 0.05). Similar differences between the mean nuclear area in colloid goiter and papillary carcinoma (124.0 + 12.27 sq microns) was also highly significant (P less than 0.001). The results obtained by image cytometry were compared with estimated DNA ploidy pattern of follicular cells from the same cases. All colloid goiter had mean nuclear area below 100 sq.microns with diploid DNA value. However, papillary carcinoma showed aneuploid DNA pattern in eight cases (80.0%), but mean nuclear area was above 100 sq.microns. A diagnostically useful finding obtained in two of the 10 cases of follicular neoplasm was the association of aneuploid DNA pattern with mean nuclear area of the follicular cells above 100 sq.microns indicating a high probability of carcinoma and thus demanding an urgent open biopsy. These cases were readily distinguished from other cases of the same category showing diploid DNA pattern and mean nuclear area of follicular cells below 100 sq.microns.

Cell Nucleus↗
PubMed1998

Phase II study of dexverapamil plus anthracycline in patients with metastatic breast cancer who have progressed on the same anthracycline regimen.

The purpose of this study is to evaluate whether metastatic breast cancer that has progressed on an anthracycline-containing drug regimen will subsequently respond to that identical regimen if dexverapamil, a modulator of P-glycoprotein-mediated drug resistance, is given concomitantly. Eligible patients received 180 mg/m2 dexverapamil every 6 h for 15 doses with the anthracycline administered 30 min after the seventh dose. Blood for dexverapamil levels was drawn before and 30 min after this dose. When possible, biopsies were obtained to measure mdr-1 expression by reverse transcription-PCR and by image cytometry. Of the 21 patients entered onto the trial, 20 were evaluable for response. There were two partial responses (10%) that both lasted for 6 months, and two additional patients had stable disease. Seven patients had asymptomatic cardiotoxicity consisting of hypotension (24%), bradycardia (5%), or prolongation of the P-R interval (14%). Two patients developed acute congestive heart failure, one on dexverapamil and one 10 days after stopping it. Dexverapamil did not seem to increase anthracycline toxicity. The median trough dexverapamil plus norverapamil level on day 3 was 1110 ng/ml (range, 186-3385 ng/ml), and the median peak level was 2164 ng/ml (range, 964-8382 ng/ml). There was poor correlation between reverse transcription-PCR and image cytometry for the level of mdr-1 expression. Because dexverapamil has been shown to affect doxorubicin pharmacokinetics subsequent to the initiation of this trial, it cannot be concluded that the responses seen were necessarily due to P-glycoprotein inhibition. Additional studies are necessary to determine whether mdr-1 modulators can reverse clinical drug resistance in breast cancer patients. The intrinsic cardiotoxicity of dexverapamil makes it less suitable for such studies than several other available agents.

ATP Binding Cassette Transporter, Subfamily B, Mem↗
PubMed1994

c-myc oncogene amplification and cytometric DNA ploidy pattern as prognostic factors in musculoskeletal neoplasms.

The relationship between c-myc oncogene amplification in neoplastic cells as determined by means of Southern-blot analysis, and their nuclear DNA content as assessed by combined flow and image cytometry, was investigated in fresh tumor specimens from 33 patients with musculoskeletal neoplasms. Amplification, without rearrangement of the c-myc proto-oncogene, was detected in 4 out of 7 bone sarcomas and in 6 out of 26 soft-tissue sarcomas, but in none of 3 benign giant-cell bone tumors. Among the 10 cases with c-myc amplification, 2 were found to be cytometrically DNA diploid, 2 DNA tetraploid, and 6 DNA aneuploid. Conversely, there were 10 tumors displaying extremely aneuploid DNA patterns without c-myc oncogene amplification. Thus, there was no relationship between c-myc amplification and DNA ploidy; neither did the percentage of S-phase cells, as determined by means of image cytometry, correlate significantly with the occurrence of c-myc amplification. A surprising sex-bias was observed; all 6 cases of c-myc-amplified soft-tissue sarcomas occurred in females, whereas none of the 11 males with such sarcomas showed this amplification. When the clinical follow-up data of the patients were scrutinized, it was found that the DNA ploidy pattern of the neoplastic cell nuclei, in combination with the S-phase values, as well as the occurrence of c-myc amplification, yielded prognostic information, being statistically significant 2 years after the diagnosis.

Adolescent↗
PubMed1994

Comparison of DNA ploidy in prostatic intraepithelial neoplasia and invasive carcinoma of the prostate: an image cytometric study.

Prostatic intraepithelial neoplasia (PIN) is regarded as the most important premalignant lesion of prostatic epithelium. The aim of this investigation was to find clues to formal pathogenesis of prostatic cancer. For this purpose DNA ploidy (determined by means of image cytometry [ICM] using 4-microns-thick Feulgen-stained paraffin sections) of PIN and invasive carcinoma was compared. Prostatic tissue of 72 patients (mean age, 67.5 years; 82 areas with carcinoma and 71 areas with PIN) was examined. In 44 cases PIN and carcinoma were coexistent in the same prostates, the PIN grade being high in 77% of these cases (P < .05). In higher-grade PIN and higher-grade carcinoma the c-values, 2.5c-exceeding-rate, and aneuploidy rate increased (P < .01). Carcinomas associated with diploid PIN (either low or high grade) showed diploidy and aneuploidy in an equal number of cases, whereas 70% of aneuploid PIN cases (all high grade) were associated with aneuploid invasive carcinomas (P < .01). Conversely, in 71% of the cases with aneuploid carcinoma the coexistent PIN (either low or high grade) was diploid. Our findings show that aneuploidy can be acquired at a preinvasive stage of carcinogenesis in the prostate and suggest that aneuploid high-grade PIN might be regarded as a precursor of some but not all aneuploid prostatic carcinomas. Image cytometry analysis seems to be a promising method for further subclassification of high-grade PIN lesions into groups with putatively lower or higher risk. However, further investigation is necessary to confirm the clinical importance of these results.

Adult↗
PubMed2003

DNA ploidy in gastrointestinal B-cell lymphomas. An image analysis study of 43 cases.

OBJECTIVE: To analyze the prognostic importance of DNA ploidy pattern on gastrointestinal (GI) B-cell lymphoma using image cytometry (ICM) and to compare the results with previously published flow cytometry (FCM) data. STUDY DESIGN: Forty-three cases of surgically resected primary GI B-cell lymphomas were examined. Thirty-eight tumors were located in the stomach, 2 in the small intestine, 1 in the large bowel and 2 in both the stomach and small intestine. Six cases were at stage E I 1, 15 at stage E I 2, 20 at stage E II 1 and 1 each at stages III and IV. Histologically, the lymphomas were classified as GI low grade marginal zone lymphoma of mucosa-associated lymphoid tissue (MALT) type (low grade, 12 cases), low grade MALT lymphoma with a high grade component (mixed type, 10 cases) and GI diffuse large B-cell lymphoma (DLBC) (high grade MALT lymphoma, 21 cases). After gross removal of nonneoplastic tissue, single cell suspensions were prepared from paraffin blocks and stained according to Feulgen. Ploidy analysis was done using a custom-made DNA cytometer and Optimas image analysis software (Optimas Corp., Seattle, Washington, U.S.A.). RESULTS: Aneuploidy was found in 42% (5/12 cases) of low grade MALT lymphoma, 90% (9/10 cases) of mixed type lymphoma and 100% (21/21 cases) of GI DLBCL. DNA ploidy had no significant impact on overall survival time (P = .73). CONCLUSION: ICM analysis showed a higher proportion of aneuploidy in GI lymphomas as compared to that in prior studies using FCM for ploidy determination. Whether DNA ploidy is an independent prognostic factor remains to be determined.

Adult↗
PubMed1993

Prognostic significance of the proliferation index in surgically resected non-small-cell lung cancer.

OBJECTIVE: To determine the utility of measuring the tumor proliferation index as a prognostic marker in patients with non-small-cell lung cancer. DESIGN: Immunostaining for the proliferation-associated antigen Ki-67, quantitated using computer-assisted image cytometry, was used to derive the tumor proliferation index for 61 fresh-frozen, banked specimens of non-small-cell lung cancer. DNA ploidy was measured concomitantly for all specimens. A median follow-up of 38 months was achieved for survival analyses. SETTING: A large southeastern United States private referral institution and affiliated hospital provided the study environment. PARTICIPANTS: A consecutive, convenience sample of 61 patients was enrolled based on resected tissue preservation and viability over a five-year accruement. MAIN OUTCOME MEASURES: Significant associations between DNA content, proliferation index, established clinicopathological parameters, and outcome were examined. RESULTS: A significant inverse association between patient survival and tumor proliferation index was found that was independent of other established clinicopathological predictors of outcome. Patients whose tumors harbored a proliferation index of less than 3.5 survived significantly longer than patients with tumors demonstrating higher values. No association between DNA content and proliferation index was uncovered. CONCLUSION: Measurement of the proliferation index, as derived from quantitative Ki-67 immunostaining and analyzed by image cytometry, may provide significant complementary, if not independent, prognostic information for patients with non-small-cell lung cancer.

Aged↗
PubMed1996

Does any correlation exist between the Gleason classification system and the computer-assisted microscope analysis of Feulgen-stained nuclei features in human prostate adenocarcinoma?

Grading prostatic adenocarcinomas remains an important problem. Various systems exist (including those proposed by Gleason) but none of these systems seems able to reliably predict either the lethal potential of a tumor in an individual patient or the responsiveness of an individual tumor to various forms of therapy. The most frequently used grading system, as proposed by Gleason, is essentially based on the description of tumor growth pattern. The aim of the present work is therefore to investigate whether the quantitative description of morphonuclear features (including cell anaplasia) and the DNA ploidy level can contribute significant information to the Gleason grading, thus partly at least reducing its subjective nature. This quantitative description was carried out by means of the Feulgen-stained nuclei image cytometry computation of 24 variables in 101 prostatic adenocarcinomas. The results show that both DNA ploidy- and morphonuclear-related variables were weak discriminators for the various grades of the Gleason classification system, and particularly between the high (Gleason 4 and 5) and the other Gleason-grades, i.e. the low (Gleason 1 and 2) and intermediate (Gleason 3) ones. The morphonuclear evidence of anaplasia is thus not redundant data on tumor growth pattern and may be expected to provide additional diagnostic information.

Adenocarcinoma↗
PubMed1996

AgNOR quantity in needle biopsy specimens of prostatic adenocarcinomas: correlation with proliferation state, Gleason score, clinical stage, and DNA content.

Aims-To define the relation between the quantity of silver stained nucleolar organiser regions (AgNORs) and histological grade, clinical stage, DNA content, and MIB-1 immunostaining in needle biopsy specimens of prostatic adenocarcinomas.Methods-Histological grade was determined according to the Gleason system. AgNOR quantity, DNA content and MIB-1 immunostaining were evaluated by image cytometry on routine histological sections stained with silver, Feulgen reaction and MIB-1 antibody, respectively.Results-The mean AgNOR area increased with increasing Gleason score. A significant difference was found in the AgNOR values between low, intermediate and high grade tumours. Patients with clinically localised tumour (stages A and B) had lower AgNOR values than patients with advanced disease (stages C and D), but the difference in the mean AgNOR values between the two groups was not statistically significant. Non-diploid tumours had a significantly higher mean (SD) AgNOR area than diploid tumours (3.68 (1.04) mum(2)v 2.73 (0.60) mum(2), respectively), while no significant difference was observed in the mean AgNOR values between aneuploid and tetraploid tumours (3.68 (1.04) mum(2)v 3.70 (1.05) mum(2)). When AgNOR and MIB-1-PI values were compared using linear regression analysis, a highly significant correlation was found.Conclusions-These data demonstrate that AgNOR quantity reflects the proliferative potential of prostatic adenocarcinomas, and is significantly related to histological grade and DNA content. The ease of application on routine sections, maintaining the morphological integrity of the tissue, the ability to evaluate selected histological areas of limited size and objective quantification by image cytometry make the AgNOR method particularly suitable for cell kinetic analysis in prostatic needle biopsy specimens.

Journal Article↗