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At least 883 records · Page 49Linked to original sources
PubMed1994

The in vitro cytokinesis-block micronucleus assay: a detailed description of an improved slide preparation technique for the automated detection of micronuclei in human lymphocytes.

An improved slide preparation technique is described for the automated detection of micronuclei in binucleate cytokinesis-blocked human lymphocytes. This automated system (Discovery Image Analyser, Becton-Dickinson Image Cytometry Systems, Leiden, The Netherlands) searches for the pattern of two touching nuclei in binucleate cells instead of the edges of the cytoplasm. For this purpose several ratios of the fixative mixture, methanol/acetic acid, were checked. After fixation with a ratio of 25:1, touching nuclei were obtained in almost 100% of the binucleate cells. A hydrolysis treatment with 5 N HCl before staining with Romanowsky-Giemsa resulted in binucleate cells with dark-stained nuclei and micronuclei and a vaguely stained cytoplasm. The high visual contrast between cytoplasm and nuclear material obtained by this staining procedure makes an accurate automated detection of micronuclei feasible. Additionally, a 64 h culture time resulted in an optimal yield of binucleate cells. The results of manual micronucleus scoring on slides prepared with the 'manual protocol' and with the 'automation protocol' indicate no significant differences between both sets of data supporting the validity of the automation protocol.

Cell Division↗
PubMed1989

Nuclear DNA content in anaplastic giant-cell thyroid carcinoma.

The nuclear DNA content was measured in 36 cases of anaplastic giant-cell thyroid carcinoma registered at Radiumhemmet, Karolinska Hospital, Sweden, between November 1973 and December 1985. The DNA content was measured in morphologically identified single tumor cells by image cytometry. Either fine needle aspirate material or histologic sections from primary tumors, recurrences, or metastases were studied. All tumors were aneuploid. Aneuploidy decreased in four of seven tumors studied sequentially. Analysis of the nuclear DNA content did not provide additional prognostic information and did not explain why occasional patients enjoy longer survival than expected.

Adult↗
PubMed2001

Serous borderline tumor of the paratestis: a report of seven cases.

We report the clinical, morphologic, immunophenotypic, and ploidy findings of seven cases of serous borderline tumor of the paratestis. Mean patient age was 56 years (range, 14-77 years), and the clinical presentation was that of a testicular mass. Tumors ranged in size from 1 to 6 cm (mean, 3.5 cm). Six tumors arose from the tunica albuginea, and two of these tumors were intratesticular. One tumor arose from the tunica vaginalis. Serous borderline tumor of the paratestis is histologically identical to its ovarian counterpart. The tumors were cystic with numerous intracystic blunt papillae lined by stratified epithelial cells with minimal to mild cytologic atypia. Psammoma bodies were present in two cases. In all cases, the neoplastic cells stained strongly and diffusely for cytokeratin 7, estrogen receptor, and CD15, and six of seven cases were positive for progesterone receptor and MOC-31. The cells did not stain for cytokeratin 20, carcinoembryonic antigen, calretinin, and HER2/neu. Proliferative activity, as assessed by MIB-1 staining, ranged from 1.3% to 10% (mean, 5.5%). Five of six tumors were diploid, and one was tetraploid. Patients were treated by radical orchiectomy and followed up from 4 months to 18 years (mean, 48 months; median, 8.5 months). No recurrences or metastases occurred. Serous borderline tumor of the paratestis is morphologically and immunophenotypically identical to ovarian serous borderline tumor. To date, no serous borderline tumor of the paratestis reported in the literature or in our series has recurred or metastasized after resection.

Adenocarcinoma↗
PubMed2003

Adenomatoid tumor of the adrenal gland: a clinicopathologic study of five cases and review of the literature.

We report the clinicopathologic, immunophenotypic, DNA ploidy, and MIB-1 proliferative findings of five adenomatoid tumors of the adrenal gland. All patients were male, and tumors were incidental radiologic, surgical, or autopsy findings. Mean patient age at diagnosis was 41 years (range 31-64 years). The tumors ranged from 1.2 to 3.5 cm (mean 2.8 cm; median 3.2 cm) in greatest dimension, and all originated within the adrenal gland. The tumors were composed of anastomosing variably sized tubules lined by epithelioid as well as flattened cells. Signet-ring-like cells were present in all cases. The previously described histologic patterns of adenomatoid tumor, adenoid, angiomatoid, cystic, and solid, were observed, and each tumor contained multiple histologic patterns. In three of five cases, there was extra-adrenal extension of tumor into periadrenal adipose tissue. All adenomatoid tumors infiltrated the adrenal cortex, and in four cases the adrenal medulla was involved. All tumors exhibited strong immunoreactivity for calretinin, cytokeratins AE1/AE3, and CAM 5.2, cytokeratin 7, and vimentin. Tumors showed weak and focal immunoreactivity for cytokeratin 5/cytokeratin 6 and were negative for CD15, CD31, CD34, cytokeratin 20, MOC31, and polyclonal carcinoembryonic antigen. Ploidy analysis using Feulgen-stained sections and image analysis showed that three tumors were diploid and two were tetraploid. Tumors exhibited low MIB-1 proliferative activity, ranging from 0.2% to 2.7% (mean 1.6%). In three cases with clinical follow-up, no recurrence or metastases occurred. Adrenal gland adenomatoid tumors are morphologically and immunophenotypically identical to adenomatoid tumors of the genital tract and appear benign.

Adenoma↗
PubMed1997

Optical imaging of low Mg(2+)-induced spontaneous epileptiform activity in combined rat entorhinal cortex-hippocampal slices.

A reproducible increase in transmission of infrared light was observed during spontaneous seizure-like events (SLEs) induced by low Mg2+ solutions in combined rat entorhinal cortex-hippocampal slices. Comparison of half maxima of transmission change in different regions indicated propagation of SLEs from the medial entorhinal cortex towards the temporal cortex suggesting spread along existing anatomical pathways. Thus, optical imaging of spontaneous epileptiform activity is possible and may improve the assessment of spread patterns. The optical signal outlasted both SLEs and associated K+ signals. In contrast, the tetraethylammonium signal, indicating changes of the extracellular space (ECS) volume, had a longer time course than the transmission changes. ECS volume changes are widely held to be responsible for transmission change. Our data suggest that other mechanisms may also contribute to increased light transmission during epileptiform activity.

Animals↗
PubMed1994

Prognostic significance of DNA ploidy in pancreatic carcinoma.

The nuclear DNA content was assessed by image cytometry on cytologic specimens obtained with fine-needle aspiration biopsy in 96 patients with pancreatic duct carcinoma. Twenty-two of these patients had pancreaticoduodenal resection. According to DNA analysis there were 27 (28%) diploid, 17 (18%) tetraploid, and 52 (54%) aneuploid tumors. The corresponding patient median survival times were 8, 5, and 4 months, respectively. Resectable tumors were more often DNA diploid (10 of 22) than nonresectable tumors (17 of 74) (p < 0.05). A Cox multivariate regression analysis demonstrated that surgical resection (p < 0.001) and diploid DNA pattern (p < 0.05) were factors associated with longer patient survival time. Although the difference in survival time was small, the data indicate that DNA diploid tumors represent a less aggressive subset of pancreatic carcinoma.

Adult↗
PubMed1997

Morphometric analysis of neointimal formation in murine cardiac allografts: II. Rate and location of lesion development.

BACKGROUND: Transplant vascular sclerosis (TVS) is manifested in transplanted human and murine hearts as a concentric, intimal lesion. The purpose of this study was to characterize the rate, location, and intensity of developing TVS lesions in murine cardiac allografts using quantitative morphometric analysis. METHODS: Murine cardiac allografts, treated with the immunosuppressant gallium nitrate, were explanted at 30, 60, and 90 days after transplant. The grafts were histologically stained and evaluated for intimal thickening by deriving a neointimal index (NI) using a computerized image-analysis system. RESULTS: In cardiac allografts, mild vascular lesions of varying NI were detectable by day 30 and lesion severity increased significantly by day 60. Thereafter, average lesion severity stabilized, although the percentage of affected vessels continued to increase from day 30 to day 90. In contrast, day-90 cardiac isografts showed little to no TVS development. Vascular lesions developed randomly without regard for vessel location or size. TVS developed more regularly in vessels of the interventricular septum than in the right or left ventricular walls. The degree of TVS development fluctuated along the length of individual vessels, even as late as 90 days after transplant. The smaller vessels (<85 microm in diameter) appeared to occlude more quickly than the larger vessels. CONCLUSIONS: TVS developed reproducibly in a random pattern throughout cardiac allografts over a 1-month to 3-month period after transplant. This development can be quantitatively monitored by computerized morphometric analysis. In general, under these experimental conditions, 30-day cardiac allografts seem to provide a useful experimental model for studying early aspects of TVS, whereas 60-day allografts may be better suited for analysis of advanced TVS.

Animals↗
PubMed1996

Diagnostic impact of nuclear DNA content and proliferative activity in benign and malignant melanocytic lesions.

Nuclear DNA content was assessed, using image cytometry, in adult melanocytes in normal skin and in 20 intradermal naevi, 60 junction naevi, 107 compound naevi, 61 dysplastic naevi, 17 melanomas in situ and 101 primary malignant melanomas. Proliferation was estimated using mitotic counting and immunohistochemical staining by anti-Ki-67 (clone MIB1) monoclonal antibodies. All normal adult melanocytes; and intradermal naevi (97% junction naevi, 98% compound naevi, 66% dysplastic naevi) were diploid. Thirty-four percent of the dysplastic naevi, 88% of the melanomas in situ and 96% of the malignant melanomas were clearly aneuploid. Proliferation, as assessed by mitotic counting and MI81 index, was significantly higher in aneuploid invasive malignant melanomas than in aneuploid dysplastic naevi (P<0.0001). The results indicate that histomorphologically defined entities of melanocytic lesions are characterized by typical DNA distribution patterns. Distinct aneuploidy combined with high proliferation rates generally seem to be well correlated to an increased malignancy potential of the lesion. In dysplastic naevi, the clonic expansion of aneuploid naevus cells may be limited by host defence mechanisms. Thus, DNA aneuploidy seems to indicate increased risk of malignant transformation, but has to be combined with other data, such as proliferation, in order to differentiate more precisely between different melanocytic lesions.

Adult↗
PubMedDate not supplied

Workshop on cancer biometrics: identifying biomarkers and surrogates of cancer in patients: a meeting held at the Masur Auditorium, National Institutes of Health.

The current excitement about molecular targeted therapies has driven much of the recent dialog in cancer diagnosis and treatment. Particularly in the biologic therapy of cancer, identifiable antigenic T-cell targets restricted by MHC molecules and the related novel stress molecules such as MICA/B and Letal allow a degree of precision previously unknown in cancer therapy. We have previously held workshops on immunologic monitoring and angiogenesis monitoring. This workshop was designed to discuss the state of the art in identification of biomarkers and surrogates of tumor in patients with cancer, with particular emphasis on assays within the blood and tumor. We distinguish this from immunologic monitoring in the sense that it is primarily a measure of the tumor burden as opposed to the immune response to it. Recommendations for intensive investigation and targeted funding to enable such strategies were developed in seven areas: genomic analysis; detection of molecular markers in peripheral blood and lymph node by tumor capture and RT-PCR; serum, plasma, and tumor proteomics; immune polymorphisms; high content screening using flow and imaging cytometry; immunohistochemistry and tissue microarrays; and assessment of immune infiltrate and necrosis in tumors. Concrete recommendations for current application and enabling further development in cancer biometrics are summarized. This will allow a more informed, rapid, and accurate assessment of novel cancer therapies.

Alternative Splicing↗
PubMed2006

Proliferation (MIB-1 expression) in oligodendrogliomas: assessment of quantitative methods and prognostic significance.

Expression of to nuclear antigen Ki-67 (MIB-1) has been linked to proliferative activity and prognosis in a variety of tumors. The authors assessed three techniques for quantitating MIB-1 (expression in oligodendrogliomas, correlating results with mitotic activity and prognosis. Formalin-fixed, paraffin-embedded sections of 38 oligodendrogliomas were immunostained using monoclonal MIB-l. Proliferation index (PI) was quantitated by visual estimation, CAS-200, and AC1S image analysis. MIB-1 expression and mitotic count were correlated with overall survival and recurrence (disease-free survival), defined clinically and radiographically as new tumor growth. Mean follow-up was 54 months (range 1-276). Mean PI quantitated by the three methods was statistically similar (Visual 10.5%, CAS-200, 12.2%, CAIS 11.2%). PI results by all three techniques correlated significantly with each other; visual and CAS-200 PI correlated with mitotic index. Overall and disease-free survivals were similar for patients with PIs above and below the mean by both image cytometric assays; visually estimated PIs below the mean, versus above the mean, correlated with improved disease-free survival. The authors show a significant correlation between MIB-1 PI using the visual method and recurrence in patients with oligodendrogliomas. The objectivity and speed of the image analysis systems make them an attractive alternative to visual estimation, and larger series should be analyzed for prognostic value.

Adult↗
PubMed1994

Proliferating cell nuclear antigen (PCNA) and p53 protein expression in Bowen's disease.

Bowen's disease is a premalignant dermatosis comprised of a clonal proliferation of atypical keratinocytes in the full thickness of the epidermis. To elucidate the relationship between the alteration of the p53 tumor suppressor protein and cell proliferation rate, we immunohistochemically examined the expression of proliferating cell nuclear antigen (PCNA) and p53 protein in 30 cases of Bowen's disease. All the cases exhibited the full-thickness distribution of PCNA-positive cycling cells in the lesional epidermis. Quantitation of PCNA staining by image cytometry revealed a mean labeling index (LI) of 75.1 +/- 20.3. p53 expression was detected in 13 cases (43%). Expression was diffuse (p53 LI > 50) in 9 cases, but focal (p53 LI < 30) in the other four. The mean PCNA LI of p53 diffusely positive cases was significantly greater than that of both p53 focally positive and p53 negative cases (89.3 +/- 10.1 vs 62.7 +/- 21.2, and 70.5 +/- 21.6; p < 0.01, respectively). These findings suggest that a high-level accumulation of p53 protein results in a more increased cell proliferation in Bowen's disease.

Aged↗
PubMed1997

Heterogeneity of DNA ploidy pattern in carcinoma of the gallbladder: primary and metastatic sites.

There are few detailed reports on the heterogeneity of the nuclear DNA ploidy pattern in carcinoma of the gallbladder. We studied twelve autopsied cases who died of extended gallbladder carcinoma. Multiple samples were taken from the primary site (Pri), from direct invasion of the liver (Hinf), from hematogenous metastasis to the liver (H), from lymphatic metastasis (LN) and from peritoneal dissemination (P). The DNA ploidy pattern was investigated by image cytometry. Heterogeneity of the DNA ploidy pattern in Pri, Hinf, H, LN and P was found in 7/11, 2/10, 5/10, 2/6 and 3/6 cases, respectively. Aneuploidy was more frequently found in Hinf than at the Pri. The DNA index of Hinf was significantly higher than that of Pri. Several stemlines, with different quantities of DNA, were found in Pri. Most of these stemlines were also observed in other sites. These facts may suggest that polyclonal cancer cells rather than one cancer cell or monoclonal cancer cells of a Pri metastasize or infiltrate, and that various polyclonal cancer cells proliferate to different degrees under different circumstances.

Aged↗
PubMed1995

DNA ploidy and vimentin expression in primary breast cancer.

The DNA content of 50 breast cancers of varying tumour type, grade and stage was measured using static image cytometry, and correlated with vimentin expression in the tumour cells. A tendency to increased vimentin expression and aneuploidy was observed in high grade and late stage tumours. A statistically significant difference was observed in DNA index and ploidy balance between grade 1 and grades 2 and 3 carcinomas (P < 0.05) and between grade I and stage II carcinomas (P < 0.05). There was a significant difference in the expression of vimentin between grades 1, 2, 3 (P < 0.001), and stages I, II, and III ductal carcinomas (P < 0.05). No significant difference was observed in the proliferation index and the degree of hyperploidy (P > 0.05) Clonal heterogeneity was observed in 25% of breast carcinomas, and was associated with increased vimentin expression. These changes may be indicative of genomic alteration and tumour aggressiveness.

Biomarkers, Tumor↗
PubMed1997

The use of an automated image cytometer for screening and quantitative assessment of cervical lesions in the British Columbia Cervical Smear Screening Programme.

The development of an automated device to screen cervical cytology slides for the detection of pre-invasive lesions of the cervix has been the goal of many individuals for over 30 years. The increasing sophistication of the technology of automation and increasingly powerful computer technology have enabled a number of these systems to reach the stage at which they have become a practical reality. The Department of Cancer Imaging at the British Columbia Cancer Agency has developed such a device over the past few years. This study reports the preliminary results of a trial to determine the reliability of the device for the screening and quantitative assessment of cervical cells. A training set of over 1000 cervical slides was used to train the image cytometer. A test set of 1030 slides was screened by the image cytometer and in the Cytology Screening Laboratory. At the 50% sample split the sensitivity of the image cytometer was 95% for severe dysplasia and 90% for moderate dysplasia, compared with a sensitivity of 90% for both of these lesions using conventional screening. A combination of nuclear texture features was found which can be used for the quantitative assessment of both abnormal cells and apparently normal intermediate cells.

Discriminant Analysis↗
PubMed2005

Predictive value of selected sperm parameters for classical in vitro fertilization procedure of oocyte fertilization.

A proportion of fertilized oocytes during classical in vitro fertilization (IVF) procedure was analysed depending on the following factors: number of mature oocytes, seminological criteria such as sperm morphology in raw semen and after its selection in a density gradient (six structural defects of a male gamete were taken into consideration), sperm concentration, motility parameters according to World Health Organization criteria and the functional tests: hypo-osmotic swelling assay and acrosomal reaction induced by calcium ionophore. Evaluation of DNA content in sperm by image cytometry and determination of malonyldialdehydes in seminal plasma were also performed. Seventy-nine semen samples from patients undergoing IVF were assessed. Apart from significant correlations obtained for selected semen parameters and proportion of fertilized eggs, logistic regression analysis showed that the best predictive factors for oocyte fertilization were normal morphology of sperm before and after gradient selection, grade B and C of sperm movement in raw semen, and DNA content after density gradient centrifugation, which all accounted for 76.7% of fertilization predictive value.

Fertilization in Vitro↗
PubMed1997

Angiomyofibroblastoma of the vulva: case report with immunohistochemical, ultrastructural and DNA ploidy studies and a review of the literature.

A case of angiomyofibroblastoma of the vulva in a 49-year-old woman was examined. The tumor measured 3 x 2.5 x 2 cm and appeared light gray to tan in color on the cut surface. Light microscopic examinations revealed that spindle or oval shaped tumor cells were arranged in loose edematous stroma with numerous thin-walled vessels. Ultrastructurally, cell organellae were not well developed but intracytoplasmic filaments of intermediate size were abundant in the tumor cells. Desmin, CD34 and vimentin immunoreactivity were detected in almost all of the tumor cells. Both estrogen and progesterone receptors were diffusely expressed in the tumor, suggestive of the sex steroid-dependency of this tumor. The KI-67 labeling index was less than 1% and the DNA content of the tumor cells, which was examined by image cytometry, demonstrated diploidy (DNA index = 0.97). These findings may reflect the quiescent or slow growing features of angiomyofibroblastoma.

Angiofibroma↗
PubMed1998

Malignant rhabdoid tumor of the vulva: case report with cytological, immunohistochemical, ultrastructural and DNA ploidy studies and a review of the literature.

A case of malignant rhabdoid tumor of the vulva in a 25-year-old female was examined. The patient presented with a subcutaneous nodule in the left labium majus. Smears of the material obtained by percutaneous fine-needle aspiration demonstrated clusters of atypical cells with prominent nucleoli. The tumor measured 6 x 5 x 5 cm and appeared tan to brown on the cut surface and partly cystic. Pathological findings obtained from intraoperative frozen tissue sections had been originally interpreted as rhabdomyosarcoma. Light microscopic examination revealed that polygonal tumor cells having vesicular nuclei with prominent nucleoli were arranged in sheets and the great majority of the tumor cells contained an eosinophilic globular paranuclear cytoplasmic inclusion. Ultrastructurally, this cytoplasmic inclusion corresponds to whirls of intermediate filaments. Vimentin immunoreactivity was detected in both the cytoplasm and cytoplasmic inclusion of almost all the tumor cells. No cytokeratin and desmin immunoreactivity were detected in the tumor cells. The Ki-67 labeling index was 36% and the DNA content of the tumor cells, which was examined by image cytometry, demonstrated diploidy (DNA index = 0.95).

Adult↗
PubMedDate not supplied

Prognostic factors in ovarian carcinoma stage III patients. Can biomarkers improve the prediction of short- and long-term survivors?

The aim of the study was to determine if biomarker expression could help discriminate between short-term and long-term survivors in women with advanced ovarian cancer. Fifty-one patients with stage III ovarian cancer were selected for the study, which included 28 short-term survivors (death from ovarian cancer within 18 months) and 23 long-term survivors (alive for more than 5 years). There was no difference between the two groups with respect to FIGO substage, age, World Health Organization score, and first-line platinum therapy. Classic clinical pathologic parameters were examined together with p53, Bcl-2, Ki-67, PDGFRalpha, P-glycoprotein, BRCA1, and DNA ploidy. Immunohistochemistry was used for scoring biomarker expression and image cytometry for DNA ploidy. All patients had primary debulking surgery followed by first-line platinum therapy. On multivariate analysis, the presence of ascites, debulking surgery and repeat laparotomy, clear-cell histology, elevated CA125, and high Ki-67 score were all found to be of prognostic importance. The long-term survivors were characterized by primary optimal cytoreduction surgery (<1 cm residual disease), attempt at maximal tumor debulking by experienced gynecological oncologic surgeons, and the absence of ascites. Normal CA125 level before platinum therapy and negative Ki-67 expression also predicted a more favorable prognosis.

Adult↗