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PubMedDate not supplied

TV-image analysis based quantification of the proliferative activity and the apoptotic rate in thyroid tumors and thyroiditis.

To analyze the impact of cellular proliferative activity and apoptosis, MIB-1 immunopositivity and the apoptotic rate of normal tissue (n = 20), follicular adenoma (n = 30), follicular carcinoma (n = 32), papillary carcinoma (n = 40), Hashimoto thyroiditis (n = 17) and de Quervain thyroiditis (n = 12) was investigated by means of TV-image analysis. Three-micron sections from paraffin-embedded surgical specimens were investigated. Immunohistochemical reactions were performed using an indirect peroxidase method. For determination of apoptosis the in situ DNA nick-end labeling method (TUNEL) was used. The rate of positive cells was determined using the CM-2 TV-image analysis system (Hund, Wetzlar, Federal Republic of Germany). Forty viewing fields (1.94 mm2) were measured with 20:1 objective mangnification. An average of > 4,400 cells were assessed in each case. The mean MIB-1 immunopositivity was higher in follicular carcinoma (average, 2.30%) than in de Quervain thyroiditis (1.48%), papillary carcinoma (1.26%), Hashimoto thyroiditis (0.97%), follicular adenoma (0.58%) and normal thyroid tissue (0.14%). The apoptotic rates were higher in Hashimoto thyroiditis (4.54%) and de Quervain thyroiditis (3.55%) than in thyroid tumors (0.31%-0.49%) and normal thyroid tissue (0.10%). Calculating the ratio of MIB-1 expression and apoptotic rate, thyroid tumors (12.1-14.6) revealed higher values than normal thyroid tissue (3.6), indicating cell gain. In Hashimoto thyroiditis (1.7) and de Quervain thyroiditis (0.7) the ratio was lower than in normal tissue indicating cell loss. MIB-1 immunohistometry and apoptotic rate quantification present new insights into the proliferative and apoptotic potential of thyroid lesions based on a high number of cells investigated per case. The impact of both methods could be used for the interpretation of diagnostically difficult and inconclusive fine-needle aspirates. However, as there was an overlap of the single values, the results have to be interpreted carefully for diagnostic purposes.

Apoptosis↗
PubMed1989

Application of expert systems to mammographic image analysis.

A prototype expert system was designed for differentiation of 16 circumscribed breast abnormalities utilizing findings identified by a human observer from X-ray mammograms, clinical data, and patient history. An off-the-shelf expert system shell served as the foundation for the rule base. The system runs on standard microcomputer hardware. Preliminary results suggest the expert system may be valuable in improving the ability of relatively unskilled observers to screen X-ray mammograms. Overall, radiology residents with limited experience in mammographic image analysis obtained a diagnostic accuracy level of 40% on their own, whereas they attained 73% accuracy level when using the expert system. A group of biomedical engineering students with no prior experience in mammographic image analysis obtained on the average an 80% diagnostic accuracy level utilizing the expert system in comparison with the 70% average accuracy level obtained by practicing radiologists without use of the system.

Expert Systems↗
PubMed1991

Quantitation of HER-2/neu oncoprotein overexpression in invasive breast cancer by image analysis: a study comparing fresh and paraffin-embedded material.

HER-2/neu oncoprotein overexpression was compared in fresh frozen and paraffin-embedded formalin-fixed invasive breast cancer material from the same patients. The HER-2/neu protein was detected by an immunohistochemical staining method, and the average amount of protein staining per cell was measured using the CAS-200 image analysis system and expressed relative to the amount of HER-2/neu protein of calibration cells of the SKBR3 cell line which are known to have amplification of the HER-2/neu gene and overexpression of the HER-2/neu protein. There was a significant correlation between degree of HER-2/neu protein overexpression and DNA-hyperdiploidy (P less than 0.01, chi 2 test). No significant correlation could be demonstrated between degree of HER-2/neu overexpression and tumor size, lymph node status, number of positive nodes or morphometric features. There was in general a good concordance (r = 0.83) in HER-2/neu expression values between fresh and paraffin-embedded material. Pairwise comparison of the two series (Wilcoxon signed ranks test) revealed no significant differences, indicating that there were no systematic differences between HER-2/neu assessments in fresh and paraffin material. When analysing the HER-2/neu expression values according to thresholds used earlier for overexpression, comparable results for fresh and paraffin material were obtained for most cases. In the fresh and paraffin material a different staining pattern was observed (more membrane staining in the fresh material in contrast to a more diffuse staining pattern in the paraffin material). It was concluded that both fresh-frozen and paraffin-embedded, formalin-fixed material is suitable for assessment of HER-2/neu protein overexpression by image analysis and provides comparable HER-2/neu expression values in most cases.(ABSTRACT TRUNCATED AT 250 WORDS)

Breast Neoplasms↗
PubMed2005

Analysis of polyomavirus-infected renal transplant recipients' urine specimens: correlation of routine urine cytology, fluorescence in situ hybridization, and digital image analysis.

Urinary polyomavirus subtype BK (BKV)-infected urothelial and renal tubular cells, "decoy cells," have been shown to be aneuploid with digital image analysis (DIA). We wanted to determine whether decoy cells cause false-positive fluorescence in situ hybridization (FISH) results in BKV-infected urine samples. Urine samples from 38 renal transplant recipients were split 3 ways and evaluated for number of decoy cells per 10 high-power fields by routine cytology, total nuclear DNA content by DIA, and chromosomal abnormalities by FISH. For DIA, Feulgen-stained cells were quantified with a CAS 200 image analyzer (Bacus Laboratories, Lombard, IL), and for FISH, the UroVysion probe set (Vysis, Downers Grove, IL) consisting of chromosome enumeration probes 3, 7, 17, and locus-specific identifier probe 9p21 (p16 gene) was used. Of the 38 specimens, 32 (84.2%) had evidence of BKV infection by routine cytology. DIA and FISH results were aneuploid/aneusomic in 30 (93.8%) and 4 (12.5%) cases, respectively. All aneusomic FISH specimens occurred in men older than 52 years of age and who also had 4 of the 6 highest PCR-BKV titers. To date, no patient has clinical evidence of malignancy. The 6 specimens without decoy cells were DNA diploid/disomic by DIA and FISH. Abnormal FISH results of urinary decoy cells occur much less frequently than aneuploidy by DIA in renal transplant recipients and might merit close follow-up in some transplant recipients.

Adult↗
PubMed2003

Angiogenesis as determined by computerised image analysis and the risk of early relapse in women with invasive ductal breast carcinoma.

The purpose of this study was to define the value of angiogenesis as a prognostic factor indicating early relapse. We assessed the relationship between parameters of angiogenesis (microvessel count--MVC, microvessel area--MVA and microvessel perimeter--MVP) and relapse-free survival at 50 months in 226 women with invasive ductal breast carcinoma. Anti CD31 antibody was used as a marker of endothelial cells. Microvessel density was measured according to Weidner et al. using a computerised image analysis. The mean parameters of angiogenesis were significantly higher in women with relapse than in those without recurrence within 50 months after surgery. In node negative patients relapse did not occur if MVC was below 38.7. In node-negative subgroup with grade II carcinomas parameters of angiogenesis in the primary tumour differed significantly depending on the presence or absence of relapse. Univariate and multivariate analysis showed the prognostic value of angiogenesis parameters in all study groups with respect to 50-month relapse-free survival. In node negative subgroup only 62.5% of women with tumors with high MVC and as much as 94.9% with low MVC survived 50 months without recurrence. In the Cox analysis of node-negative subgroup only MVC, MVP and MVA were the independent prognostic factors. In women with node-negative disease the evaluation of angiogenesis can identify a subgroup of patients with high risk of relapse, hence it may help in decisions concerning adjuvant therapy. Computerised image analysis is a good and objective technique for evaluating the intensity of angiogenesis in breast cancer.

Adult↗
PubMed1993

Distinction between diffuse cutaneous malignant follicular center cell lymphoma and lymphoid hyperplasia by computerized nuclear image analysis.

The difficult differential diagnosis between the diffuse variants of cutaneous lymphoid hyperplasia (CLH; synonym; pseudolymphoma) and malignant follicular center cell lymphomas (FCCL) often requires a multidisciplinary approach. Eighteen CLH and 11 FCCL, diagnosed by conventional histology and immunophenotyping and subsequently examined with a polymerase chain reaction to show clonal immunoglobulin heavy-chain gene rearrangements, were subjected to a novel type of automated nuclear image analysis. Of all nuclear parameters tested in azure A-stained semithin sections, the mean nuclear profile area (TN) of lymphoid cells was the best criterion to distinguish between CLH and FCCL (p = 9 x 10(-6)). Additional distinctive features, in the order of decreasing significance, were the SD of TN; all chromatin textural parameters combined; and the light and the dark fractions of the central nuclear profile areas. Parameters related to the chromatin pattern were independent of nuclear profile size in FCCL, but not in CLH. Two lesions registered as CLH displayed the nuclear characteristics favoring this diagnosis, but showed B-cell monoclonality at the DNA level. In conclusion, computerized nuclear image analysis is a helpful additional diagnostic tool in the evaluation of diffuse CLH and cutaneous FCCL.

Adolescent↗
PubMedDate not supplied

Grading follicular lymphoma on fine needle aspiration specimens. Comparison with proliferative index by DNA image analysis and Ki-67 labeling index.

OBJECTIVE: To determine whether follicular lymphoma (FL) can be graded on fine needle aspiration (FNA) biopsies by determining the percentage of centroblasts in the neoplastic follicles on the smears. STUDY DESIGN: Eighty-nine cases of histologically confirmed cases of FL, including 31 grade 1, 46 grade 2 and 12 grade 3, were evaluated. Proliferative index (PI) by DNA image analysis (DIA) and Ki-67 labeling index (LI) were obtained on all cases. A minimum of 200 cells were counted per case (range, 200-800 cells) at 40x magnification, and the number of large cells (centroblasts) was expressed as a percentage of the total number of cells counted within the follicles. RESULTS: The percentage of centroblasts in the follicular aggregates was 9.7 +/- 2.9% in grade 1 FLs, 24.7 +/- 5.6% in grade 2 and 48.4 +/- 7.5% in grade 3. These differences were significant (P < .05). DNA image analysis of PI and Ki-67 LI differed significantly between grade 1 FLs and grade 2 and 3 FLs (P < .05), but there were no significant differences between grade 2 and 3 FLs. CONCLUSION: Determining the percentage of centroblasts in the follicular aggregates on FNA specimens is a good method of grading FLs. Using the percentage of centroblasts per follicular structure, FL grades 1, 2 and 3 were adequately distinguished. PI by DIA and Ki-67 LI clearly distinguished FL grade 1 from FL grades 2 and 3; however, it did not clearly distinguish between grades 2 and 3.

Adult↗
PubMed2003

[Detection of nuclear factor kappa B by immunochemical staining and image analysis].

OBJECTIVE: To find a relatively simple and safe method for quantitative assay of nuclear factor (NF)-kappa B activity. METHODS: Immunochemical staining of NF-kappa B/p65 subunit with its antibody was performed in human umbilical vein endothelial cells stimulated with advanced glycation end products-modified human serum albumin (AGE-HSA). The ratio of p65 subunit staining in the nuclei and cytoplasm was determined by means of imaging analysis, and the results were compared with those of electrophoretic mobility shift assay (EMSA). RESULTS: NF-kappa B could be activated by AGE-HSA in a time- and dose-dependent manner, and the results obtained from immunochemical staining were consistent with those from EMSA. CONCLUSION: p65 immunochemical staining and subsequent image analysis is feasible in the quantitative detection of NF-kappa B activation.

Electrophoretic Mobility Shift Assay↗
PubMed1999

Mercury Intrusion Porosimetry and Image Analysis of Cement-Based Materials.

Mercury intrusion porosimetry (MIP) is a widely used technique for characterizing the distribution of pore sizes in cement-based materials. It is a simple and quick indirect technique, but it has limitations when applied to materials that have irregular pore geometry. The relationship between MIP results and the actual pore distribution and connectivity can be better understood with the use of image analysis. This paper discusses the use of MIP to describe the pore structure of cements and the efforts to validate the technique with microscopy. In particular, a study using molten Wood's metal as an alternate intrusion liquid that is solid in the pores at room temperature and can be examined by scanning electron microscopy will be presented. Results of the image analysis and the intrusion behavior of Portland cement mortars will be discussed. Copyright 1999 Academic Press.

Journal Article↗
PubMed1993

Nucleus image analysis as a possible prognostic tool in grading breast cancer.

The WHO grading system for breast cancer is based on the subjective, poorly reproducible evaluation of two cytological criteria (Mitotic Activity: MA and Nuclear Pleiomorphism: NP) and one histological criterion (Tubular Differentiation: TD). In order to improve the reproducibility of the assessment of tumour differentiation, we have looked for nuclear cytophotometric parameters (densitometry, geometry and texture) that could be measured objectively on smears stained by the Feulgen method. Tumour cell populations from 36 breast cancers were investigated by nuclear image analysis according to NP scores, MA scores, TD scores (ie for each variable, three categories), WHO total scores (3 to 9 or 7 categories) and WHO grades (grade I: 13 patients, whereas 13 and 10 for grade II and III respectively). Discrimination between each score or grade could be displayed by the average profiles of the nuclear cytophotometric parameters provided by multivariate analysis. Discrimination between TD scores was based on two parameters of nuclear texture. All these data suggest that WHO grading could be obtained in an objective manner by nuclear image analysis.

Adult↗
PubMed1996

Detection of baculovirus gene expression in insect cells and larvae by low light video image analysis.

Recombinant baculovirus isolates BmNPVluc and AcNPVluc (Kopylova-Sviridova et al., 1990) expressing the luc gene in Bombyx mori N-4, and in Sf 9 and Trichoplusia ni 368 cells, respectively, were studied. Luc gene expression driven by baculovirus regulatory elements was detected by enzyme and photometric assays. The expression of recombinant AcNPVluc and BmNPVluc genes in infected larvae of the cabbage looper, T. ni and the tomato hornworm Manduca sexta was analyzed by low-light video image methods. Expression of the luc gene was detected at high levels in both the lepidopteran cells and in third to fifth instar T. ni larvae. However, no light emission was detected in M. sexta caterpillars. High levels of light emission were detected in T. ni larvae when occlusion bodies containing both wild type and recombinant virus were fed to larvae. The results of these experiments demonstrate that video image analysis can be used to monitor the progression of baculovirus infection in susceptible insect cells and larvae. Bioluminescence in recombinant virus infected larvae can be used to determine virus host range, to monitor latent virus infection in insect cells and to assess the spread of recombinant viruses in the environment. Video image analysis was found to be a sensitive method for rapid detection and semiquantitative measurement of luc gene expression in baculovirus infected cells and for monitoring virus infection in larval tissues.

Animals↗
PubMed1989

Image analysis microspectroscopy of senile plaque capillary amyloid in Alzheimer's disease. A preliminary study.

The relationship between cerebral amyloid and the microvasculature in senile plaques was studied with image analysis microspectroscopy. This imaging technique permitted topographic quantification of amyloid concentration within microscopic subregions of single senile plaques. The highest concentration of amyloid was measured at the walls of intraplaque capillaries. Spectroscopic images revealed a decreasing gradient of amyloid concentration with increasing distance from intraplaque capillaries. The concentration of amyloid in nonplaque capillaries was lower than that measured in capillaries within well-developed amyloid plaques. These findings support the premise that plaque amyloid deposition starts at the level of the capillary.

Alzheimer Disease↗
PubMed2002

A simple mathematical model to aid quantification of electrophoresis gels by image analysis.

In many scientific disciplines, measurements are taken from films that have been exposed to energetic sources. Examples include radiographs where the source is an X-ray tube, autoradiography where the source is a radioactive isotope and electrophoresis gels where the source is an enhanced chemiluminescence reaction. In these situations it is of interest to quantify the darkening of the film and compute the strength of the source which in the cases of autoradiography and electrophoresis can be used to compute unknown concentrations of biochemicals. We developed a simple mathematical model of the darkening of films in radiography, autoradiography and electrophoresis bands disclosed by enhanced chemiluminescence, and present formulae to calculate the strength of the source from measurement of film blackening by image analysis. A simple model is used in two examples to predict blackening of film exposed to electromagnetic radiation. This blackening is measured by image analysis. Results show reasonable agreement between predictions of the model and blackening of film for the examples chosen. This model is proposed as an aid to quantification of electrophoresis gels.

Autoradiography↗
PubMedDate not supplied

Image analysis of nuclear/cytoplasmic ratio in cervical smears to discriminate three grades of cervical intraepithelial neoplasia.

OBJECTIVE: To suggest which system of cytologic classification (Bethesda System or World Health Organization [WHO]) of cervical lesions is more adequate for the Public Health Service of São Paulo State, Brazil, based on morphometric study with the Jandel Videoanalysis (JAVA) System. STUDY DESIGN: The study comprised groups of typical smears: cervical intraepithelial neoplasia (CIN 1) separated into two subgroups: cytopathic effects of human papillomavirus associated or not with dyskaryosis, CIN 2 and CIN 3. The JAVA system of image analysis was used to determine the nuclear/cytoplasmic (N/C) ratio in abnormal cells from each group. RESULTS: Significant differences were detected between the three grades of CIN and between the two subgroups of CIN 1. CONCLUSION: Although image analysis is not applicable to large-scale population screening of cervical smears, its use in the present study favored the three-grade cytologic classification (WHO). In addition, the three-grade classification offers the clinician more options for treatment. Considering the clinical-laboratory characteristics of our public health service, the three-grade classification is more adequate.

Cell Nucleus↗
PubMed1989

Image analysis of transparent particle systems by mathematical morphology methods.

The particle images are often distorted by apparent holes due to undeflected transmitted light. These apparent holes must be disregarded for a consequent image analysis. The reconstruction of the real particles by methods of mathematical morphology was used. The problems associated with touching particles were also dealt with. The method is illustrated on an ion-exchanger system.

Algorithms↗
PubMed2000

Microregional heterogeneity of non-protein thiols in cervical carcinomas assessed by combined use of HPLC and fluorescence image analysis.

Under low oxygen conditions, non-protein thiols (NPSHs, non-protein sulfhydryls) can effectively compete for DNA radicals sites and hence represent a potentially important cause of radiation resistance in the clinic. Intra- and intertumoral heterogeneity of glutathione (GSH) and cysteine were assessed in cryostat sections of multiple biopsies obtained from 10 cervical carcinomas by the combined use of a sensitive high-performance liquid chromatography (HPLC) method and a fluorescence image analysis technique to examine the spatial distribution of NPSHs in tumor tissue. Glutathione concentrations ranged from 1.98 to 4.42 mM; significant (> or =1 mM) concentrations of cysteine, a more effective radioprotector than GSH, were found in some tumors. By HPLC, the intratumoral heterogeneity of NPSHs was relatively small compared with the intertumoral heterogeneity. The histochemical stain 1-(4-chloromercuryphenoylazo)-2-napthol (mercury orange), which binds to GSH and cysteine, was used to determine the spatial distribution of NPSHs in tumor tissue. A comparison of NPSH levels in serial cryostat sections showed a close correlation between NPSH values determined by HPLC and mercury orange fluorescence quantification. Using fluorescence image analysis, an approximately 2-fold increase of NPSHs in tumor versus nonmalignant tissue was observed in the same section. Because some cervical carcinomas contain radiobiologically important levels of cysteine, agents that target the biochemical pathways maintaining tumor cysteine have therapeutic potential as adjuncts to radiotherapy in cervix cancer patients.

Adenocarcinoma↗
PubMed1990

[Centroblast morphology: a morphometric evaluation by means of computer-aided image analysis].

In cytologic preparations, cell images from 30 cases of high grade malignant Non-Hodgkin's lymphoma (NHL) and 10 cases of tonsillitis have been analysed using a colour TV microscope system, high resolution scanning and image processing. Multivariate analysis of 30 cell features allowed satisfying recognition and discrimination of centrocytes, centrocyte-like centroblasts, ordinary centroblasts and multilobated centroblasts. In addition, according to their image analytical data, the ordinary centroblasts from NHL formed 5, and those from tonsillitis 4 well defined, homogeneous clusters. Even at simultaneous analysis, there was an essential overlap only between centroblast subtype 3 from NHL and subtype 9 from tonsillitis, but reliable discrimination of all the rest.

Computers↗
PubMed2004

Photoshop-based image analysis of canine articular cartilage after subchondral damage.

INTRODUCTION: The validity of histopathological grading is a major problem in the assessment of articular cartilage. Calculating the cumulative strength of signal intensity of different stains gives information regarding the amount of proteoglycan, glycoproteins, etc. Using this system, we examined the medium-term effect of subchondral lesions on initially healthy articular cartilage. MATERIALS AND METHODS: After cadaver studies, an animal model was created to produce pure subchondral damage without affecting the articular cartilage in 12 beagle dogs under MRI control. Quantification of the different stains was provided using a Photoshop-based image analysis (pixel analysis) with the histogram command 6 months after subchondral trauma. RESULTS: FLASH 3D sequences revealed intact cartilage after impact in all cases. The best detection of subchondral fractures was achieved with fat-suppressed TIRM sequences. Semiquantitative image analysis showed changes in proteoglycan and glycoprotein quantities in 9 of 12 samples that had not shown any evidence of damage during the initial examination. Correlation analysis showed a loss of the physiological distribution of proteoglycans and glycoproteins in the different zones of articular cartilage. CONCLUSION: Currently available software programs can be applied for comparative analysis of histologic stains of hyaline cartilage. After subchondral fractures, significant changes in the cartilage itself occur after 6 months.

Animals↗