Malignancy-associated Changes Research Articles

Histological Findings in Kidney Biopsies of Patients with Monoclonal Gammopathy-Always a Surprise.

Background: The simultaneous occurrence of impaired kidney function and paraproteinemia is common in our constantly aging society. Both can be independent entities; however, renal insufficiency can also be caused by the paraprotein. We assessed all kidney biopsies in patients with monoclonal gammopathy in our clinic over the past 20 years and evaluated the histological results. Methods: Biopsies were systematically performed in nearly all patients with paraproteinemia and impaired kidney function (n = 178). The histological findings were systematically evaluated and correlated with the initial clinical diagnosis. Results: We found cast nephropathy (CN) in n = 66 (37.1%) biopsies, AL amyloidosis in n = 31 (17.4%) biopsies, monoclonal immunoglobulin deposition disease (MIDD) in n = 7 (3.9%) biopsies and other renal diseases (ORDs) in n = 74 (41.6%) biopsies. In the latter group, paraprotein-associated changes were found in 37 of 74 (50%) patients, whereas paraprotein-independent changes were found in the other half. Whereas, in the group of patients with MGUS, the findings were heterogenous, most of the patients with known multiple myeloma (MM) or B-NHL showed malignancy-associated changes in the kidney. The biopsy changed the diagnoses in a significant proportion of the patients: The group of patients with MM grew from 71 to 112 patients, whereas, in the MGUS group, only 31 of 44 patients remained. Conclusion: Kidney biopsies in patients with paraproteinemia and renal impairment show a wide range of findings that can lead to a change in diagnosis.

Methodology for structuring a dataset of cervical cell images for the study of malignancy changes associated in conventional Pap Test

The Pap test is a screening procedure used worldwide for the diagnosis of cervical cancer. The MalignancyAssociated Changes (MAC) are slight morphological and textural changes that take place in the chromatin ofthe cell nucleus. Their study allows the early detection of several types of cancers. However, the number ofdata set for MAC studies is limited by the technical aspects that must be followed. In this work, we createda data set for the study of MAC in images of cervical cells from conventional cytology exams. To extract thecharacteristics of the nuclei, the color channels were separated by the contribution of Hematoxylin and Orange G6dye, which identified morphological, illumination and texture characteristics. As a result, 1186 nuclei of patientseither with some type of anomaly or without anomalies were obtained. To separate the regions of the nuclei, thenon-linear regression function and theWatershedalgorithm with an accuracy of 85.6 % were used. This dataset contains adequate data to complement future work in MAC study of cervical cells from conventional pap smears.

Spectral histopathology of the lung: A review of two large studies.

This paper summarizes results from two large lung cancer studies comprising over 700 samples that demonstrate the ability of spectral histopathology (SHP) to distinguish cancerous tissue regions from normal tissue, to differentiate benign lesions from normal tissue and cancerous lesions, and to classify lung cancer types. Furthermore, malignancy-associated changes can be identified in cancer-adjacent normal tissue. The ability to differentiate a multitude of normal cells and tissue types allow SHP to identify tumor margins and immune cell infiltration. Finally, SHP easily distinguishes small cell lung cancer (SCLC) from non-SCLC (NSCLC) and provides a further differentiation of NSCLC into adenocarcinomas and squamous cell carcinomas with an accuracy comparable of classical histopathology combined with immunohistochemistry. Case studies are presented that demonstrates that SHP can resolve interobserver discrepancies in standard histopathology.

P2.11-14 Malignancy Associated Change and The LuCED® Test for Detection of Early Stage Lung Cancer

Early detection remains the most reliable and effective strategy for curing lung cancer. Many approaches, however, are limited by poor sensitivity or specificity that increase health care costs and potentially risk patient health through unneeded procedures. The association between cell morphology and cancer has been established in the pathology literature. However, through the so-called field effect, cancer can introduce subtle morphological changes into non-cancerous cells that are proximal to the tumor site. The Cell-CT™ platform and LuCED® test represent a promising new method for detecting lung cancer with high (92%) sensitivity and (95%) specificity based on cytologically abnormal cells. In this research we investigate use of the Cell-CT to detect malignancy associated changes in normal cells near the cancer with a view towards enhancing LuCED test performance. We present a study of 3D morphological alterations in non-cancer cells obtained from sputum of healthy subjects and biopsy confirmed lung cancer patients. Three major cell types were analyzed from 235 patients: bronchial epithelial columnar, squamous intermediate, and mature macrophages. We used the Cell-CT™ platform to measure over 700 different structural biomarkers for each cell. The measurements were used to define prominent clusters of cells through a hierarchical process that were then used under supervised learning, with case status as ground truth, to create classifiers that optimally separated cells from cancer vs. normal cases. The table gives classifier development and performance characteristics:Tabled 1Cell TypeNumber of cells from cancer patientsNumber of cells from normal patientsCluster-based Supervised Learning – aROCSquamous Intermediate23166840.94Macrophages496050400.94Columnar cells322732340.92 Open table in a new tab Our results indicate that the Cell-CT can discriminate cell features that are too subtle to distinguish by a human. The study suggests that detection of cells with Malignancy Associated Changes may be used to further enhance the LuCED test’s performance beyond published levels.

Identification of Malignancy-Associated Changes in Histologically Normal Tumor-Adjacent Epithelium of Patients with HPV-Positive Oropharyngeal Cancer

The incidence of HPV-positive oropharyngeal cancer (HPV+ OPC) is increasing, thus presenting new challenges for disease detection and management. Noninvasive methods involving brush biopsies of diseased tissues were recently reported as insufficient for tumor detection in HPV+ OPC patients, likely due to differences between the site of tumor initiation at the base of involuted crypts and the site of brush biopsy at the crypt surface. We hypothesized that histologically normal surface epithelial cells in the oropharynx contain changes in nuclear morphology that arise due to tumor proximity. We analyzed the nuclear phenotype of matched tumor, tumor-adjacent normal, and contralateral normal tissues from biopsies of nine HPV+ OPC patients. Measurements of 89 nuclear features were used to train a random forest-based classifier to discriminate between normal and tumor nuclei. We then extracted voting scores from the trained classifier, which classify nuclei on a continuous scale from zero (“normal-like”) to one (“tumor-like”). In each case, the average score of the adjacent normal nuclei was intermediate between the tumor and contralateral normal nuclei. These results provide evidence for the existence of phenotypic changes in histologically normal, tumor-adjacent surface epithelial cells, which could be used as brush biopsy-based biomarkers for HPV+ OPC detection.

In vivo Raman spectroscopy of oral buccal mucosa: a study on malignancy associated changes (MAC)/cancer field effects (CFE)

Occurrence of metachronous and synchronous secondary tumors in oral cavities has been associated with poor prognosis and decreased 5-year disease-free survival rates. The origin of secondary tumors in the oral cavity has been primarily attributed to cancer field effects (CFE) or malignancy-associated changes (MAC) in uninvolved areas. Classification of normal, cancerous and pre-cancerous oral lesions by in vivo Raman spectroscopy (RS) has already been demonstrated. In the present study, MAC/CFE in oral buccal mucosa were explored. In vivo Raman spectra from 84 subjects (722 spectra) under five categories - cancer and contralateral normal (opposite side of tumor), healthy controls (no tobacco habit, no cancer), habitués healthy controls (tobacco habit, no cancer) and non-habitués contralateral normal (no tobacco habit with cancer) were acquired. Mean and difference spectra suggest that loss of lipids and additional features representing proteins and DNA are characteristics of all pathological conditions, with respect to healthy controls. Spectral data were analyzed by PC-LDA followed by leave-one-out cross-validation. Results suggest that Raman characteristics of mucosa of healthy controls are exclusive, while those of habitués healthy controls are similar to those of contralateral normal mucosa. It was observed that the cluster of non-habitués contralateral normal mucosa is different from habitués healthy controls, suggesting that malignancy associated changes can be identified and also indicating that transformation of uninvolved oral mucosa due to tobacco habit or malignancy is different. The findings of the study demonstrate the potential of RS in identifying early transformation changes in oral mucosa and the efficacy of this approach in oral cancer applications.

Precursors of prostate cancer

High-grade prostatic intraepithelial neoplasia (PIN) is the only accepted precursor of prostatic adenocarcinoma, according to numerous studies of animal models and man; other proposed precursors include atrophy and malignancy-associated changes (with no morphologic changes). PIN is characterized by progressive abnormalities of phenotype and genotype that are intermediate between benign prostatic epithelium and cancer, indicating impairment of cell differentiation and regulatory control with advancing stages of prostatic carcinogenesis. The only method of detection of PIN is biopsy because it does not significantly elevate serum prostate-specific antigen concentration and cannot be detected by ultrasonography. The mean incidence of PIN in biopsies is 9% (range, 4%-16%), representing about 115,000 new cases of isolated PIN diagnosed each year in the United States. The clinical importance of PIN is its high predictive value as a marker for adenocarcinoma, and its identification warrants repeat biopsy for concurrent or subsequent carcinoma, especially when multifocal or observed in association with atypical small acinar proliferation (ASAP). Carcinoma develops in most patients with PIN within 10 years. Androgen deprivation therapy and radiation therapy decrease the prevalence and extent of PIN, suggesting that these forms of treatment may play a role in prevention of subsequent cancer. Multiple clinical trials to date of men with PIN have had modest success in delaying or preventing subsequent cancer.

Cytopathology by optical methods: spectral cytopathology of the oral mucosa

Spectral cytopathology (SCP) is a novel approach for diagnostic differentiation of disease in individual exfoliated cells. SCP is carried out by collecting information on each cell's biochemical composition through an infrared micro-spectral measurement, followed by multivariate data analysis. Deviations from a cell's natural composition produce specific spectral patterns that are exclusive to the cause of the deviation or disease. These unique spectral patterns are reproducible and can be identified and used through multivariate statistical methods to detect cells compromised at the molecular level by dysplasia, neoplasia, or viral infection. In this proof of concept study, a benchmark for the sensitivity of SCP is established by classifying healthy oral squamous cells according to their anatomical origin in the oral cavity. Classification is achieved by spectrally detecting cells with unique protein expressions: for example, the squamous cells of the tongue are the only cell type in the oral cavity that have significant amounts of intracytoplasmic keratin, which allows them to be spectrally differentiated from other oral mucosa cells. Furthermore, thousands of cells from a number of clinical specimens were examined, among them were squamous cell carcinoma, malignancy-associated changes including reactive atypia, and infection by the herpes simplex virus. Owing to its sensitivity to molecular changes, SCP often can detect the onset of disease earlier than is currently possible by cytopathology visualization. As SCP is based on automated instrumentation and unsupervised software, it constitutes a diagnostic workup of medical samples devoid of bias and inconsistency. Therefore, SCP shows potential as a complementary tool in medical cytopathology.

Abstract A10: Gene expression analysis of malignancy associated changes in normal bronchial epithelium

Abstract A10 Background Lung cancer is the number one cancer-related cause of death in the world. The poor prognosis for lung cancer patients is largely due to the lack of an effective non-invasive method to detect the disease in its early stage. Using quantitative nuclear morphometry of sputum cells, we have previously demonstrated that in the presence of tumour, cytologically normal appearing diploid cells may exhibit malignancy-associated changes (MACs) and that these changes may be used as surrogate markers for early detection. The molecular basis for MACs is not known. Objective Our objective is to identify malignancy-associated gene expression changes in normal bronchial epithelial cells obtained by bronchial brushing. Methods We compared the pre- and post-surgical resection gene expression profiles of bronchial epithelial cells obtained by brushing of airways in the lung opposite to the one containing the lung cancer. Agilent whole genome expression arrays were used to generate mRNA transcript profiles for seven paired normal bronchial brushings taken before and 3 to 6 months after surgical resection of the tumour. Ingenuity Pathway Analysis software was used to identify downstream expression targets of the soluble signaling factors VEGF, TGF-β, TNF-α, and IL-8, which have been reported to be secreted by lung tumour cells. The target genes identified were analyzed for malignancy induced differential gene expression between normal bronchial epithelial cells from cancerous and non-cancerous environments. Results We detected differential expression of multiple target genes in the bronchial brush cells. Genes that were found to be differentially expressed included downstream targets of the signaling molecules EGF,TGF-β, and VEGF. Conclusion Soluble signaling molecules secreted by lung malignancies induce MACs, as evident by altered gene expression after removal of the tumour. Although further investigation is necessary, our results are consistent with previous observation that lung tumours can induce MACs in normal appearing bronchial cells in the vicinity of the tumor. MACs may be a potential marker for early detection of lung cancer. This data also provides evidence that MACs can be measured quantitatively in bronchial brush cells using gene expression analysis. Citation Information: Cancer Prev Res 2008;1(7 Suppl):A10.

Mechanisms of Disease: high-grade prostatic intraepithelial neoplasia and other proposed preneoplastic lesions in the prostate

High-grade prostatic intraepithelial neoplasia (HGPIN) is the most likely precursor of prostatic adenocarcinoma according to virtually all available evidence. This lesion is characterized by cellular proliferations within pre-existing ducts and acini, with nuclear and nucleolar enlargements similar to those seen in prostate cancer, although unlike cancer HGPIN retains a basal-cell layer. The recognition of HGPIN is clinically important because of the strong association between this disease and prostatic carcinoma. The predictive value for cancer of an initial diagnosis of HGPIN on needle biopsy has substantially declined, with values falling from 36% to 21%. A major factor contributing to this decline is related to increased use of needle biopsy core sampling, which has provided the means for many cancers associated with HGPIN to be detected on initial biopsy; repeat biopsy, even with good sampling, does not detect many additional cancers. Other possible findings in the prostate might indicate premalignant disease (low-grade prostatic intraepithelial neoplasia, atrophy, malignancy-associated changes, and atypical adenomatous hyperplasia or adenosis), but the data for these premalignant diseases are much less convincing than those for HGPIN.

Epigenetic events, remodelling enzymes and their relationship to chromatin organization in prostatic intraepithelial neoplasia and prostatic adenocarcinoma

To explore the nuclear chromatin phenotype, overall epigenetic mechanisms, chromatin remodelling enzymes and their role as diagnostic biomarkers in prostate lesions, using high-resolution computerized quantitative digital image analysis (DIA). A tissue microarray (TMA) was constructed using paraffin wax-embedded prostatic tissues from 78 patients, containing 30 cores of benign prostatic hyperplasia (BPH), 10 of low-grade prostatic intraepithelial neoplasia (LGPIN), 38 of prostate adenocarcinoma, 20 of BPH tissue excised at 0.6-1 mm from LGPIN lesions, and 10 of BPH prostatic tissues obtained 0.6-1 mm from high-grade PIN (HGPIN) lesions. Chromatin phenotype was assessed on haematoxylin-stained sections using high-resolution texture analysis. For quantitative immunohistochemistry, antibodies raised against acetylated histone H3 lysine 9 (AcH3K9), 5'methylcytidine (5MeC) and the chromatin remodelling ATPase ISWI (SNF2H and SNF2L) were used. The immunodensity was measured using DIA to determine the epigenetic profile of the cases. At least 60 nuclei were measured from each case. There were many statistically significant differences in staining intensity and nuclear distribution patterns in chromatin phenotype and immunostaining (p </= 0.001). These changes allowed the differentiation between the various pathological subgroups with a classification accuracy of 76-100% using chromatin phenotype or immunostaining measuring epigenetic and chromatin remodelling changes (5MeC, AcH3K9 and ISWI). In PIN lesions, there was a high chromatin content with DNA-hypermethylation, while in prostatic adenocarcinoma there was a lower chromatin content with DNA-hypomethylation and H3K9-hypoacetylation. There was significantly more ISWI protein in neoplastic tissues. There were malignancy-associated changes (MAC) in chromatin phenotype and overall epigenetic events in BPH tissues adjacent to PIN lesions. The present study confirms the ability of high-resolution computerized digital imaging of nuclear texture features to detect changes in chromatin phenotype, epigenetic events and the presence of chromatin remodelling, factors that can be used to distinguish between different prostatic pathologies, i.e. BPH, LGPIN, HGPIN and prostate adenocarcinoma, and further allow the detection of MAC near PIN lesions. These results provide a base for future diagnostic applications of DIA combined with immunohistochemistry. Our experiments underscore the importance of epigenetic mechanisms during carcinogenesis. Further studies are needed to elucidate the complex interplay between chromatin structure, its modifications and the progression of prostate cancer.

Thrombosis in multiple myeloma

Multiple myeloma, as with other malignancies, has been associated with the development of venous thromboembolic events. Chemotherapy or steroids in combination with antiangiogenic agents can further enhance this risk. The identification of measurable factors associated with this prothrombotic state could help in the selection of patients who need antithrombotic prophylaxis. Malignancy-associated thrombophilic state, paraprotein-specific mechanisms and treatment-induced changes can explain the high rate of thrombosis in this cancer population. While the release of inflammatory cytokines induces high levels of factor VIII, von Willebrand factor and downregulate the protein C system, elevated plasma immunoglobulin can impair fibrinolysis. Strategies of thromboprophylaxis with low molecular weight heparin, warfarin or aspirin in patients treated with thalidomide/chemotherapy or lenalidomide and dexamethasone have shown efficacy. Early data indicate that the effect of low molecular weight heparin on multiple myeloma is not confined to the anticoagulant effect but could extend to survival; a similar positive trend in overall survival has also been reported in patients treated with aspirin.

Criteria for Malignancy in Gastrointestinal Endocrine Tumors

In contrast with the large amount of data generated from endocrine tumors of the pancreas, sparse and mostly unconfirmed data are available on the criteria for the assessment of malignancy risk and patient outcome in endocrine tumors of the gastrointestinal tract. In these conditions the 2000 WHO classification with its standardized scheme of pathologic report constitutes a framework facilitating the assessment of tumor malignancy and has been regarded as useful for clinical purposes, providing the basis for proper management of the patients and for the design of treatment protocols. The classification is based on a combination of pathological and clinical features with parameters specific for each organ in which the endocrine tumors originate. Three main categories, one further subdivided into two subgroups, are considered: (1) well-differentiated endocrine tumors, further subdivided into tumors with benign and with uncertain behavior; (2) well-differentiated endocrine carcinomas, low grade; and (3) poorly differentiated endocrine carcinomas, high grade. In this review the differential tumor characteristics between the different categories are summarized. Moreover, the relevance of additional features with respect to tumor prognostication, chiefly the Ki-67 proliferation index and malignancy-associated genetic changes, is discussed with emphasis on the discrepancies emerging between tumors of foregut and of midgut origin.

Exploratory analysis of quantitative histopathology of cervical intraepithelial neoplasia: objectivity, reproducibility, malignancy-associated changes, and human papillomavirus.

As part of a project to evaluate emerging optical technologies for cervical neoplasia, our group is performing quantitative histopathological analyses of biopsy specimens from 1,190 patients. Objectives in the interim analysis are (a) quantitatively assessing progression of the neoplastic process of cervical intraepithelial neoplasia (CIN)/squamous intraepithelial lesions (SIL), (b) detecting malignancy-associated changes (MACs), and (c) phenotypically measuring human papillomavirus (HPV) detected by DNA testing. The diagnostic region of interest (ROI) from immediately adjacent sections were imaged, and the basal lamina and surface of the superficial layer were delimited. Nonoverlapping quantitatively stained nuclei were selected from 1,190 samples with histopathological characteristics of normal (929), koilocytosis (130), CIN 1 (40), CIN 2 (23), and CIN 3/carcinoma in situ (CIS) (68). A fully automatic procedure located and recorded the center of every nucleus in the region of interest (ROI). We used linear discriminant analysis to assess the changes between normal and CIN 3/CIS. Scores computed from the cell-by cell features and the clinical grade of CIN/SIL were highly correlated, as were those of the architectural features and the clinical grade of CIN/SIL. We found even higher correlations between a combination of cell-by-cell and architectural scores, and clinical grade. Using these scores, we found MACs in the normal biopsy specimens from patients with high-grade CIN/SIL. Furthermore, the same scores correlated with the molecular detection of HPV. Quantitative histopathology can be used in large clinical trials as an objective and reproducible measure of CIN/SIL. Detectable phenotypic changes correlate well with CIN/SIL neoplastic progression. It can also be used to infer the presence of CIN/SIL (MACs) and molecular changes associated with increased risk of cancer development (high-risk HPV).

In vitro model for studying malignancy associated changes.

Malignancy associated changes (MAC) can be defined as subtle morphological and physiologic changes that are found in ostensibly normal cells of patients harboring malignant disease. It has been postulated that MAC have a potential to become a useful tool in detection, diagnosis and prognosis of malignant diseases. An in vitro cell culture model system was designed to study interactions between non‐small cell lung cancer (NSCLC) and the normal bronchial epithelium of the human respiratory tract in vivo to see if the MAC‐like phenomenon can be detected in such a system. In this study we examined changes in nuclear features of normal human bronchial epithelial cells (NHBE) when they were co‐cultured with cells derived from a lung cancer cell line NCI‐H460. Using discriminant function analysis, nuclear features were determined which allow maximal discrimination between normal cells incubated with or without cancerous cells. Our results demonstrate that MAC appear to be specific to changes induced by malignancy, and that these changes differ from those induced by growth factors in the serum. This study provides evidence in support to the hypothesis that MAC are induced by a soluble factor(s) released by malignant cells. Colour figure can be viewed on http://www.esacp.org/acp/2003/25‐2/sun.htm.

Malignancy Associated Changes (MAC) in the lungs resected due to NSCLC

Malignancy Associated Changes (MAC) in the lungs resected due to NSCLC

Malignancy associated changes in bronchial epithelial cells and clinical application as a biomarker

A total of 74 bronchial brushing specimens, 24 from patients with advanced stage cancer, eight from patients with CIS, 31 from patients with atypical metaplasia and 11 from normal subjects were examined for the existence of malignancy associated changes (MAC). Conventional fiberoptic bronchoscopy and fluorescence endoscopy was carried out on every case. Each case was classified according to the highest grade of abnormality diagnosed by bronchial biopsy of the suspect areas. During the endoscopy examination, a bronchial brushing specimen was obtained from a visually normal area very remote from the abnormal area as possible such as the opposite lung or another lobe. The bronchial brushing specimens were fixed, mounted and stained by a DNA specific method and approximately 1500 images of individual nuclei per case were captured by an automated high resolution image cytometry. For each of these images, more than 100 nuclear features such as size, shape and chromatin spatial organization were calculated. Discriminant function analysis revealed nuclear features which differentiated between normal bronchial cell nuclei from the normal subjects and ostensively normal nuclei (MAC cell nuclei) from the lung cancer patients. The best discrimination was achieved when the frequency of individual cells expressing MAC was 50% or greater. With this threshold, 75% of the patients with invasive cancer and CIS were correctly classified. Fifty percent of those with severe or moderate atypia and 35% with mild atypia were also MAC positive. The frequency of cells expressing MAC also increased as the degree of abnormality of the groups increased. MAC may be a useful criterium to determine biological behavior of the intra-epithelial (pre-invasive) neoplasia.

Subtle changes in benign tissue adjacent to prostate neoplasia detected with a Bayesian belief network.

The aim of this paper was to test the usefulness of a Bayesian belief network (BBN) as a decision support system in the uncertainty assessment of benign prostatic tissue, either associated or not with inflammation or adjacent to prostatic adenocarcinoma (PAC) or prostatic intraepithelial neoplasia (PIN). A shallow network was used with eight first-level descendant nodes for the diagnostic clues, each independently linked by a conditional probability matrix to a root node containing the diagnostic alternatives. One diagnostic evidence node was based on the tissue architecture and the others were based on cell features. The efficacy of the network was tested on a series of 45 simple prostatectomy specimens, subdivided as follows; benign prostatic tissue not associated with other diseases (15 cases), associated with acute and/or chronic inflammation (15 cases), and adjacent to accidentally discovered PAC or PIN (15 cases). The highest belief values for the diagnostic alternative normal prostate (NP) were obtained in the 15 cases not associated with other diseases, the mean value being 0.996. The 15 cases evaluated in areas with inflammation showed the lowest belief values for NP (mean 0.774). For the 15 cases evaluated in specimens with PAC or PIN, the belief values for NP were intermediate between those from normal prostatic tissue associated with inflammation and those not associated (mean 0.925). Moreover, it was found that subtle changes were also present at a certain distance from the tumour. In conclusion, the network can be used as a decision support system to differentiate with high certainty benign prostate adjacent to PAC or PIN from benign prostatic tissue either associated or not with inflammation. The subtle morphological alteration detected with the BBN may be considered malignancy-associated changes.

Disease type-associated increases of the plasma levels and ligand expression for natural alpha- or beta-galactoside-binding immunoglobulin G subfractions in patients with lung cancer

The presence of natural carbohydrate-binding antibodies may play a role in host defence against malignant cells in addition to elicitation of an immune response by artificial carbohydrate antigens. Human serum contains immunoglobulin G(2) (IgG) fractions with selectivity to alpha- and to beta-galactosides, respectively, irrespective of the type of blood group of the donor. To determine whether these naturally occurring subfractions may have any relevance for tumor disease control, their binding to malignant cells was ascertained by cytofluorimetric assays in vitro with a number of human tumor cell lines of different histogenetic origin. The affinity of cell binding was comparable to that of binding to lactosylated or melibiosylated neoglycoconjugates as model ligands in solid-phase assays and K-D values were found to be in the range of 5-300 nM. Cross-reactivity of the anomer-selective subfractions to the other type of ligand was observed to be rather low. When the IgG contents of plasma samples of patients with diverse types of lung cancer were assessed, the concentrations of both galactoside-binding immunoglobulin G subfractions were significantly increased in association with presence of small cell lung carcinoma and of metastatic lesions to the lung without any marked change in the overall IgG plasma level. Such an apparently general enhancement was seen for patients with adenocarcinoma and included both subfractions with no impact on their percentage in the total IEC content. When detergent extracts of tumor and tumor-free specimens of the same patient were analyzed with the affinity purified antibody subfractions to comparatively determine ligand presentation, increases in sugar-inhibitable binding were especially noted for the tumor tissue of small cell lung carcinomas and apparently tumor-free samples of cases with lung metastasis. Material from other types of lung cancer revealed no significant indication for disease-related alterations with the exception of carcinoids. These data demonstrate that plasma levels and ligand expression for two types of natural galactoside-binding immunoglobulin G fractions can show nonuniform responses in patients within the class of lung cancer. They encourage to deliberately monitor these parameters of the natural carbohydrate-directed antibody fractions in cancer patients with various types of disease to clarify the clinical significance of respective malignancy-associated changes.

Detection of malignancy associated changes in cervical cell nuclei using feed-forward neural networks.

Normal cells in the presence of a precancerous lesion undergo subtle changes of their DNA distribution when observed by visible microscopy. These changes have been termed Malignancy Associated Changes (MACs). Using statistical models such as neural networks and discriminant functions it is possible to design classifiers that can separate these objects from truly normal cells. The correct classification rate using feed‐forward neural networks is compared to linear discriminant analysis when applied to detecting MACs. Classifiers were designed using 53 nuclear features calculated from images for each of 25,360 normal appearing cells taken from 344 slides diagnosed as normal or containing severe dysplasia. A linear discriminant function achieved a correct classification rate of 61.6% on the test data while neural networks scored as high as 72.5% on a cell‐by‐cell basis. The cell classifiers were applied to a library of 93,494 cells from 395 slides, and the results were jackknifed using a single slide feature. The discriminant function achieved a correct classification rate of 67.6% while the neural networks managed as high as 76.2%.