DNA Karyometry for Automated Detection of Cancer Cells.

Background: Microscopical screening of cytological samples for the presence of cancer cells at high throughput with sufficient diagnostic accuracy requires highly specialized personnel which is not available in most countries. Methods: Using commercially available automated microscope-based screeners (MotiCyte and EasyScan), software was developed which is able to classify Feulgen-stained nuclei into eight diagnostically relevant types, using supervised machine learning. the nuclei belonging to normal cells were used for internal calibration of the nuclear DNA content while nuclei belonging to those suspicious of being malignant were specifically identified.

Automated detection of cancer cells in effusion specimens by DNA karyometry.

Background: The average sensitivity of conventional cytology for the identification of cancer cells in effusion specimens is only approximately 58%. DNA image cytometry (DNA-ICM), which exploits the DNA content of morphologically suspicious nuclei measured on digital images, has a sensitivity of up to 91% for the detection of cancer cells. However, when performed manually, to our knowledge to date, an expert needs approximately 60 minutes for the analysis of a single slide.

Removing defocused objects from single focal plane scans of cytological slides.

Background: Virtual microscopy and automated processing of cytological slides are more challenging compared to histological slides. Since cytological slides exhibit a three-dimensional surface and the required microscope objectives with high resolution have a low depth of field, these cannot capture all objects of a single field of view in focus. One solution would be to scan multiple focal planes; however, the increase in processing time and storage requirements are often prohibitive for clinical routine.

Comparability of tumor-cytogenetics and -DNA-cytometry.

Knowledge of the karyotypes of cancer cells is the theoretical underpinning of diagnostic/prognostic DNA-cytometry. Currently the translation is hampered by different terminologies of both fields. The following letter tries to close current gaps between the two fields by harmonizing their different terminologies.

Is it useful to combine sputum cytology and low-dose spiral computed tomography for early detection of lung cancer in formerly asbestos-exposed power industry workers?

Background: Low-dose spiral computed tomography (LDSCT) in comparison to conventional chest X-ray proved to be a highly sensitive method of diagnosing early stage lung cancer. However, centrally located early stage lung tumours remain a diagnostic challenge. We determined the practicability and efficacy of early detection of lung cancer when combining LDSCT and sputum cytology.

Equivocal cytology in lung cancer diagnosis: improvement of diagnostic accuracy using adjuvant multicolor FISH, DNA-image cytometry, and quantitative promoter hypermethylation analysis.

Background: Sometimes, cytological lung cancer diagnosis is challenging because equivocal diagnoses are common. To enhance diagnostic accuracy, fluorescent in situ hybridization (FISH), DNA-image cytometry, and quantitative promoter hypermethylation analysis have been proposed as adjuncts.

Methods: Bronchial washings and/or brushings or transbronchial fine-needle aspiration biopsies were prospectively collected from patients who were clinically suspected of having lung carcinoma.

Role of brush biopsy and DNA cytometry for prevention, diagnosis, therapy, and followup care of oral cancer.

Late diagnosis resulting in late treatment and locoregional failure after surgery are the main causes of death in patients with oral squamous cell carcinomas (SCCs). Actually, exfoliative cytology is increasingly used for early detection of oral cancer and has been the subject of intense research over the last five years. Significant advances have been made both in relation to screening and evaluation of precursor lesions.

Hemodynamic support by left ventricular assist devices reduces cardiomyocyte DNA content in the failing human heart.

Background: Whether adult cardiomyocytes have the capacity to regenerate in response to injury and, if so, to what extent are still issues of intense debate. In human heart failure, cardiomyocytes harbor a polyploid genome. A unique opportunity to study the mechanism of polyploidization is provided through the setting of hemodynamic support by left ventricular assist devices.

Toward a multimodal cell analysis of brush biopsies for the early detection of oral cancer.

Background: This report describes what to the authors' knowledge is the first clinical application of semiautomated multimodal cell analysis (MMCA), a novel technique for the early detection of cancer for cases with a limited number of suspicious cells. In this clinical study, MMCA was applied to oral cancer diagnostics on brush biopsies. The MMCA approach was based on the sequential application of multiple stainings of identical, slide-based cells and repeated relocalizations and measurements of their diagnostic features, resulting in multiparametric features of individual cells.

Core classification of lung cancer: correlating nuclear size and mitoses with ploidy and clinicopathological parameters.

We attempted to establish a microscopy based tumour characterization providing insight into the genetics of cancer cells and in particular their DNA ploidy. The core classification defined semi-quantitative criteria for scoring the nuclear size ranging from small (core score 1) to giant nuclei (core score 4). By listing all nuclear sizes according to their relative frequencies it provided a measure for core size variability as a correlate of nuclear pleomorphism.

Immunocytochemical typing of primary tumors on fine-needle aspiration cytologies of lymph nodes.

The aim of this study was to analyze the role of immunocytochemistry as an ancillary method on routine FNACs of enlarged lymph nodes, using different markers. In a validating cohort study all patients had confirmatory histological and/or clinical follow-up. 10 FNACs were analyzed for the differentiation of Non-Hodgkin Lymphoma (NHL) from metastatic carcinoma (MC), 30 cases to identify the sites of metastatic unknown primary tumors and 16 cases were checked to confirm clinical suspicion of a specific MC.

9p21 Deletion in the diagnosis of malignant mesothelioma in serous effusions additional to immunocytochemistry, DNA-ICM, and AgNOR analysis.

Background: The diagnosis of malignant mesothelioma (MM) in serous effusions is difficult but may be achieved by the application of adjuvant methods.

Methods: The authors cytologically diagnosed 33 effusions as suspicious or positive for MM cells by using DNA-image cytometry (DNA-ICM), immunocytochemistry and AgNOR analysis. The authors further detected 9p21 deletions by chromosomal fluorescence in situ hybridization (FISH).

Early cytological diagnosis of diffuse malignant mesothelioma of the peritoneum: a case report.

Mesotheliomas are the most frequent primary malignant tumors of serosal cavities with a poor prognosis. A definitive and early diagnosis on effusion samples is important, because recent advances in therapy for patients with mesothelioma may result in an improved outcome if they are applied to stage I disease. We report a case of malignant peritoneal mesothelioma diagnosed repeatedly by cytology in ascites fluids 1.

[Minimally invasive brush-biopsy: innovative method for early diagnosis of oral squamous cell carcinoma].

The aim of this prospective and blinded study was to investigate the diagnostic accuracy of conventional cytopathology of oral brush biopsies taken from suspicious oral lesions. In addition we checked slide based DNA image cytometry as an adjuvant diagnostic tool. Our hypothesis is that DNA aneuploidy is a sensitive and specific marker for earliest detection of oral cancer using brush biopsies.

Additional use of DNA-image cytometry improves the assessment of resection margins.

Background: Despite the histopathologic findings of tumor-free margins, patients with oral squamous cell carcinoma (SCC) often suffer from local tumor relapse. The purpose of this study was to determine the prognostic value of DNA-image cytometry in the assessment of resection margins.

Methods: DNA-image cytometry was performed in 40 SCC patients with histologically tumor-free resection margins.

Immunocytochemical diagnosis of hepatocellular carcinoma and identification of carcinomas of unknown primary metastatic to the liver on fine-needle aspiration cytologies.

Background: Difficulties with cytologic diagnoses on fine-needle aspiration cytology (FNAC) of the liver can be overcome by the application of immunocytochemical panels applied on smears. The aim of the current study was to analyze the performance of a panel of monoclonal antibodies to differentiate hepatocellular carcinoma (HCC) from metastatic carcinoma (MC) or regenerative nodules, and to identify the to date unknown primary sites of carcinomas that had metastasized to the liver.

Methods: In a validating cohort study, 108 FNACs coin lesions in the liver were routinely evaluated applying immunocytochemistry as an ancillary method.

Methylation of RAS association domain family protein 1A as a biomarker of lung cancer.

Background: Promoter hypermethylation is an important mechanism for silencing tumor-suppressor genes in cancer and a promising tool for development of molecular biomarkers. This study aimed to determine the prevalence of RAS association domain family protein 1A (RASSF1A) promoter hypermethylation in bronchial aspirates of patients with suspected lung cancer and to test whether this type of methylation assay could be used as a diagnostic adjunct to conventional cytology.

Methods: Two hundred three bronchial aspirates from patients with suspected lung cancer were analyzed for RASSF1A hypermethylation by using a sensitive quantitative methylation-specific polymerase chain reaction (QMSP).

Methylation assay for the diagnosis of lung cancer on bronchial aspirates: a cohort study.

Purpose: Recent studies have detected aberrant promoter methylation of adenomatous polyposis coli promoter 1 A (APC), cyclin-dependent kinase inhibitor-2A (p16(INK4a)), retinoic acid receptor beta2, and RAS association domain family protein 1 (RASSF1A) in bronchial aspirates and suggested their use as biomarkers for lung cancer diagnostics. The purpose of this study was to validate these candidate marker genes in a retrospective cohort study.

Experimental Design: Bronchial aspirates collected from a cohort comprising 247 patients with suspected lung cancer were investigated retrospectively regarding aberrant promoter methylation using a quantitative methylation-specific real-time PCR (QMSP).

Immunocytochemical identification of carcinomas of unknown primary in serous effusions.

Metastases from carcinomas of unknown primary site (CUP) in serous effusion are a common clinical problem. Immunocytochemistry was applied as an adjunct to the cytological diagnosis of metastatic carcinomas in serous effusions. Subjects of this study were 118 pleural, 53 peritoneal, and 9 pericardial effusions from 180 patients routinely investigated in the Institute of Cytopathology.

Cytologic and DNA-cytometric follow-up of oral leukoplakia after CO2- and Er:YAG-laser assisted ablation: a pilot study.

Background And Objectives: The aim of the present pilot study was to determine therapeutic responses to Er:YAG- and CO(2)-laser ablation in patients with oral leukoplakia as evaluated by means of exfoliative cytology (EC) and DNA-image-cytometry (DNA-I).

Study Design/materials And Methods: Ten patients exhibiting a total of 16 lesions affecting a variety of intraoral sites were randomly treated with either (1) an Er:YAG laser (300 mJ/pulse, 10 Hz, defocused mode) (ERL), or (2) an CO(2) laser (4-6 W, 20-50 Hz, focused mode) (CO). Brush (B) and incisional (I) biopsies were obtained from the respective lesions immediately before treatment (B, I) as well as 24-96 weeks postoperatively (B).

Aberrant promoter methylation of p16(INK4a), RARB2 and SEMA3B in bronchial aspirates from patients with suspected lung cancer.

Aberrant promoter methylation of normally unmethylated CpG-islands offers a promising tool for the development of molecular biomarkers. We investigated bronchial aspirates of patients admitted for suspected lung cancer with regard to the prevalence of aberrant methylation of potential marker genes. Applying quantitative methylation specific PCR (QMSP) we analyzed bronchial aspirates from 75 patients with primary lung cancer and 64 bronchial aspirates of patients diagnosed with benign lung disease for promoter methylation of 3 candidate marker genes (p16(INK4a), RARB2 and SEMA3B).

Identification of progressive cervical epithelial cell abnormalities using DNA image cytometry.

Background: The objectives of the current study were to compare the capabilities of conventional cervical cytology and of DNA image cytometry (DNA-ICM) in the prediction of progressive or regressive behavior in atypical squamous cells (ASC), low-grade squamous intraepithelial lesions (LSIL), and atypical glandular cells (AGC).

Methods: One hundred ninety-six women with Papanicolaou (Pap) smears that yielded diagnoses of ASC, LSIL, or AGC were included in a prospective cohort study. Slides were classified according to the Bethesda system.

Diagnostic and prognostic value of Ki67 proliferation fraction in serous effusions.

The Ki67 proliferation rate of mesothelial cells was determined in 20 effusions due to malignant mesotheliomas and in 20 non-neoplastic effusions, to investigate if this marker may be useful to identify neoplastic mesothelioma cells and if there is a correlation between proliferation rate and survival time. Using the ABC-method, effusions were immunostained and the marker Ki67 was evaluated quantitatively. Ki67 proliferation fraction showed rates from 2.