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PubMed1995

Gastric carcinoma: recent issues in prognostic factors.

The prognosis of gastric carcinoma remains unfavorable despite a greater understanding of its molecular pathology. This retrospective study of primary gastric carcinomas was collected from one of the highest risk regions of China and examined for the oncogenetic expression of p53, c-erbB-2, and PCNA using immunohistochemistry and DNA contents by flow cytometry and image analysis. These products are reported to influence the tumor behavior. The p53 nuclear and c-erbB-2 membrane-bound stainings were seen in 58% and 34% of cases, respectively. A high PCNA index was found in 90% of the tumors. The p53 expression did not correlate with the histological differentiation, gross morphology, and depth of tumor invasion. Additionally, p53 and c-erbB-2 reactivity did not correlate with the proliferative index (PI) or S-phase DNA content. However, the mutant p53 expression was detected in the dysplastic cells adjacent to the tumor, suggesting a possible role of the oncogene in tumor pathogenesis. Mutant p53 expression can also be helpful in early detection of cases with dysplasia in well-differentiated adenocarcinomas.

Adenocarcinoma↗
PubMed1995

[Predictive value of cytology for the efficacy of chemotherapy in breast cancers].

For the past decade, primary chemotherapy has been extensively used in breast cancer treatment. However, valuable indicators of tumor response are needed. Fine-needle cytopuncture allows the study of malignant cells by cytologic examination, image analysis and flow cytometry. Some parameters evaluated before treatment (cytologic grade, nuclear area and S phase) or at the beginning of treatment (cytomorphologic and cell kinetic changes) have been shown to be correlated with tumor response. The methods, their value and limitations, and results of the literature are discussed.

Antineoplastic Combined Chemotherapy Protocols↗
PubMedDate not supplied

Lead poisoning due to hai ge fen. The porphyrin content of individual erythrocytes.

A 45-year-old Korean man developed abdominal colic, muscle pain, and fatigue. Following a 3-week hospitalization, acute intermittent porphyria was diagnosed based on the symptoms and a high level of urinary delta-aminolevulinic acid (378 mumol/L [4.95 mg/dL]). However, discovery of an elevated blood lead level (3.7 mumol/L [76 micrograms/dL]) subsequently led to the correct diagnosis. No occupational source of lead exposure was identified. The patient reported ingesting a Chinese herbal preparation for 4 weeks prior to becoming ill. A public health investigation revealed that the source of lead exposure was hai ge fen (clamshell powder), one of the 36 ingredients of the Chinese herbal medicine. We used fluorescence image-based cytometry to determine the frequency distribution of the zinc protoporphyrin content in circulating red blood cells and found that 70% of the patient's cells contained elevated levels of zinc protoporphyrin, consistent with the duration of lead exposure and effect of lead on heme synthesis. Analysis of zinc protoporphyrin content in circulating red blood cell distributions may be useful in the diagnosis, therapy, and kinetic modeling of lead poisoning. Environmental lead poisoning is best addressed through the close collaboration of clinicians, public health specialists, and laboratory scientists.

Animals↗
PubMed1993

Effectiveness of pulse-shape criteria for the selection of dicentric chromosomes by slit-scan flow cytometry and sorting.

A method was developed to detect dicentric chromosomes by slit-scan flow cytometry. The two centromeres of dicentric chromosomes are represented by the two dips in the trimodal fluorescence profile. A trimodal profile can, however, also be generated by aggregates of chromosomes. We tested the effectiveness of slit-scan profile criteria that were applied to discriminate between trimodal profiles generated by dicentrics and trimodal profiles generated by artefacts. A Profile-Dip Counter (PDC) module was designed that can assess, in real time, the number of dips in slit-scan profiles. The PDC module was used in combination with a Cytofluorograph System 50 cell sorter for slit-scan sorting of chromosomes prepared from irradiated V79 cells. Chromosomes corresponding to trimodal profiles were sorted individually onto slides for subsequent visual inspection by fluorescence microscopy. The isolated chromosomes were stretched by treatment with trypsin to increase the efficiency for centromere detection. When fixed with glutaraldehyde, chromosomes could be sorted intact on slides. We found that trimodal profiles are generated by dicentric chromosomes as well as by monocentric and aggregated chromosomes. When stringent pulse-shape criteria were applied for the selection of profiles, the yield of dicentric chromosomes was 70% of the sorted chromosomes.

Animals↗
PubMed1995

Diagnostic and prognostic decision support systems.

Diagnostic decision support systems provide a quantitative evaluation of diagnostic evidence and the capability to combine diagnostic evidence in such a manner that a numeric measure of certainty in a final diagnostic recommendation results. Generally, expert systems serve to establish a diagnostic decision, inference networks allow a detailed analysis of the diagnostic value of diagnostic clues, case-based reasoning systems are designed to provide a prognostic assessment targeted to an individual patient. In all of these systems, symbolic information, i.e., traditional diagnostic, linguistic terms and concepts are processed and quantitatively evaluated.

Decision Support Techniques↗
PubMed1995

Clinical applications of Bayesian belief networks in pathology.

Bayesian belief networks (BBNs) are a novel tool for representing knowledge about diagnostic decision making and for obtaining a numerical measure of certainty in the final diagnosis. Belief networks have been applied to the pathological assessment of breast, prostate and skin lesions and have been shown to provide consistency in the grading of microscopic features and improve diagnosis. These applications are reviewed in the current paper. The application of BBNs has been further facilitated through the use of standardised imagery which is stored digitally and used to enter evidence into a BBN. It is predicted that the further development of BBNs with improved logical capabilities represent the key to improved decision making in pathology.

Bayes Theorem↗
PubMed1995

Syntactic structure analysis.

Syntactic structure analysis is a relatively new technique in diagnostic quantitative pathology that, based on the graph theory, can provide quantitative information on tissue architecture. This article gives an introduction to syntactic structure analysis and discusses the potential of syntactic structure analysis in diagnostic quantitative pathology. In addition, some examples of its application in histo- and cytopathology are presented and it is speculated which other applications the user of syntactic structure may find for quantitation of architecture in diagnostic quantitative pathology.

Algorithms↗
PubMed1995

Organized quantitative pathology. Short review of the activities of the Committee for Diagnostic Quantitative Pathology from 1981 to the foundation of the International Society of Diagnostic Quantitative Pathology in 1994.

The Committee for Diagnostic Quantitative Pathology (CDQP), known originally as the Committee for Diagnostic Morphometry, ceased to exist under the original title and was converted to the International Society for Diagnostic Quantitative Pathology (ISDQP) in Amsterdam, September 14, 1994. The history of this society started in 1981 in a conference < > held at Koli, Finland. Since the original meeting the group of quantitative pathologists organized yearly gatherings: Symposia on Diagnostic Quantitative Pathology (earlier known as Symposia on Morphometry in Morphological Diagnosis) every other year, and meetings in association with the European Society of Pathology Congresses in the intervening years. In 1981, the symposium had 23 participants, in 1994, the International Society for Diagnostic Quantitative Pathology had over 300 members from six continents. During the short period of its existence the society has witnessed a steadily growing trend in educational courses on quantitative pathology. The general policy of the Committee, now Society, has willingly supported all activities which can be expected to lead to valuable results in quantitative microscopy and associated fields either through development of education, methodology or practical applications. By arranging a course of Diagnostic Quantitative Pathology the society participates in the activities of the European School of Pathology in Torino. The next symposia of the Society will be arranged in Heidelberg, October 1995 and in Sendai, Japan, October/November 1996.

Europe↗
PubMed1995

Age and experimental obstructive emphysema. A morphometrical study on the rat.

Age, as a risk factor in the development of experimental obstructive emphysema, is proposed as the hypothesis of this study. Ninety-two Wistar rats were organized into two age groups: adult (16 weeks) and middle-aged (56 weeks). Each age group was subdivided into three groups: a control group, consisting of unmanipulated animals; a "cannula" group consisting of animals into whose trachea a cannula was implanted; and a "valve" group, consisting of animals into whose trachea a valve had been implanted. The survival was one month. A histomorphometric study was performed on the lungs and the results were compared statistically. Throughout the experiment the amount of food consumed by each animal and the variations in weight were monitored. After sacrifice, the lungs were processed for light microscopy. Thirteen histomorphometric variables were quantified and subsequently systematized into three groups: those which quantified the size of the distal airspace ("area of the alveolar section", "alveolar chord" and "mean linear intercept"): those which quantified the tissue ("wall thickness", "tissue density", "internal perimeter of each alveolar section", "internal alveolar perimeter per field" and "alveolar section/section perimeter"); and those which quantified the elastic fibre ("elastic fibre area", "elastic fibre perimeter", "elastic fibre area/elastic fibre perimeter", "elastic fibre density" and "elastic fibre density per tissue density"). The results were compared statistically and the sensitivity, specificity and misclassification indices were calculated, as well as the attributable and relative risk. From the results, it was observed that, in general, the animals of the valve and cannula groups in both age groups displayed a decrease in food intake and a body weight loss. The middle-aged animals were the only group which displayed significant differences in all the morphometric variables except wall thickness, when the cannula and valve groups were compared with the control group. In both the cannula and valve groups, the values of the variables which quantified the distal airspace increased, while the values of the variables which quantified the lung tissue and the elastic fibre decreased. In the manipulated middle-aged group, the attributable risk of developing emphysema was 56.66% and the relative risk 5.55; in the group of manipulated adult animals, the attributable risk was 23.55% and the relative risk 1.66. The results of this study lead us to propose that the middle-aged rats with experimental airflow obstruction displayed a greater risk of developing emphysema than the adult rats which were subjected to the same procedure.

Aging↗
PubMed1995

Histometry and the H.O.M.E. concept: an aid to the grading of intra-cervical neoplasia?

The view has been expressed that few quantitative methods are of value to the pathologist in purely diagnostic work. Quantitative systems are perceived as too large for the average reporting room, too time consuming to learn, very expensive to buy and quick to become obsolete. Further, the software supplied usually cannot provide fully automated analysis, and user interaction is often tedious. If measurement techniques have little value in diagnosis they may have a role in assessing the prognosis of tumours. High levels of inter- and intra-observer variation in tumour grading have been reported and quantitative methods have been used to reduce this and more emphasis has been placed on the measurement of changes in tissue architecture, which may help to reduce observer variation. This paper describes such a method based on cell sociology, which has been implemented on a quantitative microscope specifically designed for use in the routine diagnostic pathology environment. The results of a preliminary study on grading cervical intraepithelial neoplasia show a significant difference between all groups (P less than 1 x 10(-5)) and a linear trend for the measurement of Area Disorder (P less than 1 x 10(-5)).

Female↗
PubMed1996

Proliferative activity and p53 expression in adenoid cystic carcinoma of the breast.

Adenoid cystic carcinoma is a rare type of invasive breast carcinoma that has a good prognosis. We studied a series of four cases of adenoid cystic carcinoma in which we correlated the clinical and pathological features. The pathological features examined included light microscopy; electron microscopy; immunohistochemistry using antibodies to keratin, vimentin, S100 protein, actin, estrogen and progesterone receptors, and proliferation marker MiB-1, and p53 suppressor protein; image cytometric analysis for measurement of DNA ploidy; and molecular analysis using polymerase chain reaction single strand conformation polymorphism to assess point mutation of the p53 gene. All of the cases had a low nuclear grade, were negative for estrogen and progesterone receptors, and were DNA diploid. Three of the cases showed no evidence of metastases and had small primary tumors with low proliferative activity and absence of p53 protein expression. In contrast, one of the cases showed axillary lymph node metastases and in this case the primary tumor was large with a higher proliferative activity and expression of p53 protein, suggesting that these factors might play a role in the biological behavior of adenoid cystic carcinoma.

Adult↗
PubMed1996

DNA image analysis of urinary cytology: prediction of recurrent transitional cell carcinoma.

To evaluate the utility of image analysis in monitoring patients with transitional cell carcinoma, we studied, by cytologic means and by image analysis, 78 urinary tract specimens from 66 patients, of whom 49 (74%) had a previous history of transitional cell carcinoma. The specimens consisted of 51 (65%) voided urine specimens, 12 (15%) bladder washings, 8 (10%) ureteral washings, 3 (4%) ureteral brushings, 2 (3%) renal pelvic washings, and 2 (3%) catheterized urine specimens. DNA histograms were classified into five patterns on the basis of their DNA index and the percentage of their cells with DNA content greater than 5c: diploid (single peak in the 2c region with no cells greater than 5c), intermediate (diploid with less than 10% of cells greater than 5c), aneuploid (single peak or multiple peaks between the 2c and 4c region or more than 10% of cells greater than 5c), tetraploid (at least 10% of cells in the 4c region and a corresponding peak at 8c), and polyploid (multiple peaks in the 2c, 4c, 8c, and 10c regions). Of the 78 cases, 22 were diploid, 24 were intermediate, 29 were aneuploid, one was tetraploid, and two were polyploid. Histologic confirmation or clinical follow-up was found in 29 aneuploid cases, 13 intermediate cases, and one diploid case. Most cases of carcinoma in situ (five of six) and invasive tumors (12 of 17) were aneuploid. The sensitivity was 100%, and the specificity was 73% when cytologic and image analysis results were combined. We conclude that image analysis, when combined with cytologic examination, is a reliable noninvasive diagnostic test for monitoring patients with transitional cell carcinoma; aneuploidy is specific for malignancy; and the presence of cells greater than 5c, although frequently associated with tumor recurrence, can be seen in non-neoplastic conditions.

Adult↗
PubMedDate not supplied

Quantitative determination of thymic eosinophilia in swine.

Thymuses from 512 slaughtered pigs weighing about 160 Kilos, were subjected to histological examination. All thymuses displayed a focal or diffuse eosinophilia in the connective tissue septa and in the lymphoid tissue, especially in the medulla. Quantitative determination of cellular density and eosinophils was performed on 30 cases, by image analysis, and the number of eosinophils was compared to the number of Hassall's bodies. Statistical analysis demonstrated a significant difference in the number of eosinophils between cases with focal eosinophilia and those with diffuse pattern (P < 0.001). The higher quantity of eosinophils was present, the higher number of Hassall's bodies was calculated per unit area of medulla (P < 0.001). The significance of this eosinophilia is obscure. It may be related to the elimination of cellular catabolism products and this agrees with the possible phagocytic activity of Hassall's bodies. However, further investigation concerning the physiopathology of the thymus must take into consideration the presence of the eosinophils granulocytes in this organ.

Animals↗
PubMed1996

Cell cycle expression of p53 protein, c-Myc gene product and tyrosine-phosphorylation level determined by image analysis in human breast cancer cells.

OBJECTIVE: To investigate the cell cycle expression of p53 protein, c-myc gene product and tyrosine phosphorylation level in human breast cancer cells. STUDY DESIGN: Using a multifluorescence imaging procedure, the concentration per cell in different phases of the cell cycle can be evaluated by analyzing the bivariate contour plot of DNA content versus antigen concentration. RESULTS: Low fluorescence intensity was observed in the G0/G1 phase for the three markers. The analysis of individual cells demonstrated that approximately 10% of cells were negative. During the G1/S transition, the fluorescence intensity of the three antigens increased rapidly. However, after the mild S-phase, the increase of c-myc was more marked than the tyrosine phosphorylation level, whereas p53 protein remained stable, with a slight tendency to decrease. CONCLUSION: This study confirmed that the p53 protein and c-myc gene product could perform a regulatory function in G1/S transition and, consequently, may play an important role in malignant transformation. Like-wise, the variations of tyrosine kinase activity were linked to cellular progression throughout the cell cycle and could be a useful marker of alteration in the growth-factor signaling pathway. Thus, the multifluorescence imaging procedure may provide useful information on the mechanisms of the cell cycle and on malignant transformation.

Antibody Specificity↗
PubMed1996

Interferon therapy, but not busulfan restores normal-sized megakaryopoiesis in CML--a comparative histo- and immunomorphometric study.

To assess possible alterations of megakaryocytes associated with interferon (IFN) and busulfan (BU) therapy of Ph(1+)-CML, an immunohistochemical and morphometric study was performed on trephine biopsies of the bone marrow taken before and at varying intervals during treatment. For the identification of megakaryopoiesis and its endoreduplicative activity the monoclonal antibodies CD61 (anti-platelet glycoprotein IIIa) and PC10 raised against proliferating cell nuclear antigen (PCNA) were used. We compared 60 specimens from 20 patients following IFN alpha-2b administration (in combination with IFN gamma in seven patients) with 57 specimens from 22 patients after monotherapy with BU. A close correlation with clinical follow-up studies revealed that in the IFN-treated group the prevalence of atypical micro-megakaryocytes, usually characterizing CML, was conspicuously reduced in repeatedly taken bone marrow samples. Initially, even an increase in size which was levelled to normal values during maintenance therapy was observed. These features were most prominently expressed in the 13 patients with a complete hematologic and/or partial cytogenetic response. Associated with this phenomenon was a significant enhancement of the PCNA-labelling index which indicated a stimulation of endoreduplicative (endomitotic) activity necessary for achieving normal size and ploidy. In the second group of patients treated by BU these changes were absent. For this reason, our findings are in keeping with the assumption that during IFN treatment, there is at least partial recovery and expansion of a putative normal (Ph1-) megakaryopoiesis. In conclusion, megakaryocyte morphology, i.e. normalization in size, is thought to be a useful indicator to evaluate the response to IFN in CML patients.

Adult↗