Eosin-stained Paraffin Sections Research Articles

New method of nuclear grading of tissue sections by means of digital image analysis with prognostic significance for node-negative breast cancer patients.

To optimize treatment of the individual patient with node-negative breast cancer, objective, reproducible, and standardized prognostic criteria are required. A number of factors have been studied in recent years, but until now it has been possible to obtain information about the risk of recurrence only for some patients belonging to subgroups with special characteristics. We report the establishment of an image analysis method for nuclear grading as an attempt to solve this problem. In a retrospective analysis, we used routine hematoxylin and eosinstained paraffin sections from 54 node-negative patients with surgery between 1980 and 1985. Cell scenes of primary tumors were scanned in a light microscope in successive focus positions to obtain three-dimensional information. After automatic image segmentation, nuclear features were calculated as input for a first binary classification tree to differentiate between tumor and nontumor cells. Tumor nuclei from patients with or without relapse were defined as high-risk or low-risk nuclei, respectively, and were separated with a second tree. Feature values of the measured tumor nuclei from each patient were examined with this second tree to analyze whether the majority of nuclei for each patient were classified as high-risk or low-risk nuclei. Correct classification rates in the two binary cell classification trees were 88.0% and 83.8%, respectively. In the learning sample of our study, all patients with a relapse had the majority of nuclei in the high-risk group, most with more than 80%. Therefore, it seems to be possible to develop an image analytical risk profile system for nuclear grading to provide information on individual prognosis.

Morphometric Distinction of Sclerosing Adenosis from Tubular Carcinoma of the Breast

Objective features have been identified that assist in distinguishing sclerosing adenosis from tubular carcinoma of the breast. Hematoxylin and eosin stained paraffin sections of 18 sclerosing adenoses and 18 tubular carcinomas were studied using a TAS Plus image analysis system. Histological measurements from lumens and glands included stereologic features of architecture and morphometry of size and shape (the latter by Fourier coefficients). Cytological measurements included nuclear area, perimeter, diameter and formfactor. Initial analysis suggested utility for several individual features. However, after a modified Bonferroni procedure only two of the features were statistically significant, i.e. the coefficient of variation of luminal form factor and the surface density of glands. Multivariate discriminant analysis using these two variables correctly classified 86% of the cases, with three adenoses and two carcinomas misclassified. Validity of the discriminant rules was supported by classification using measurements from a separate, independent set of cases (ten sclerosing adenoses and nine tubular carcinomas). The classification function computed from the first set misclassified only one case from the second set, a tubular carcinoma, leaving 95% of the cases successfully categorized. Chi square test for 2 x 2 contingency tables gave a p-value < 0.001 for both sets of cases. The results suggest that morphometric features are helpful in distinguishing tubular carcinoma from sclerosing adenosis and point out the need for conservative analysis of high-dimensional data sets.

Intestinal microsporidiosis as a cause of diarrhea in human immunodeficiency virus-infected patients: A report of 20 cases

Chronic diarrhea accompanied by weight loss is a common and often debilitating problem associated with human immunodeficiency virus (HIV) infection. Enterocytozoon bieneusi, a newly identified species of the phylum of protozoa, Microspora, has been reported associated with chronic diarrhea and wasting in 11 acquired immunodeficiency syndrome (AIDS) patients in the United States, Europe, and Africa. Diagnosis has been based solely on the ultrastructural identification of this small, intracellular parasite in bowel biopsies. Seventy-one small bowel biopsies from 67 homosexual AIDS and AIDS-related complex patients with chronic diarrhea and with no pathogens identified by light microscopy on paraffin sections, were embedded in plastic and studied by light and transmission electron microscopy. Enterocytozoon bieneusi microsporidiosis was diagnosed by electron microscopy in 20 (22 biopsies) of the patients. More jejunal biopsies (16 of 36) were positive than duodenal biopsies (six of 35). Parasites and spores were clearly visible at the light microscopic level in the semi-thin plastic sections from 17 and 21 of the biopsies, respectively. In retrospect, parasites could be identified by light microscopy in standard hematoxylin and eosin-stained paraffin sections. Infection was confined to enterocytes covering the villi, especially the tips, and was associated with villous atrophy and cell degeneration, necrosis, and sloughing. Release of spores into the bowel lumen was evident. Colorectal biopsies from two of the patients with small bowel microsporidiosis were negative for microsporidia. Enterocytozoon bieneusi infection of the small bowel may be an important cause of diarrhea in HIV-infected persons.

Effects of unilateral naris closure on the olfactory epithelia of adult mice

This study demonstrates, for the first time, that prolonged unilateral breathing can be harmful to the adult olfactory epithelium. Mice at least 5 months old had one naris closed by cautery and suture. These were divided into 5 groups of 10 mice which had unilateral naris closure for 1, 3, 6, 8 or 12 weeks. A control group of 10 mice was untreated. Variables that were assessed included the thicknesses and numbers of cells spanning the olfactory epithelia in hematoxylin and eosin stained paraffin sections. Olfactory marker protein (OMP) immunohistochemistry was used to further visualize the differential impact of naris closure on the two sides of the nose. Unilateral naris closure for 6 weeks or longer caused dramatic losses of olfactory receptor cells in the rostral third of the open-side olfactory epithelia, but did not affect numbers of cells in caudal regions or on the closed sides. The thicknesses of the open and closed-side olfactory epithelia were significantly different for only the 8-week closure group. In most mice with unilateral naris closure for longer than 6 weeks there was little or no staining of the olfactory receptor neurons or their axon bundles for OMP in the affected regions of the open side.

Prenatal ethanol exposure and cerebellar development in rats

Rats of Sprague-Dawley strain were fed a liquid ethanol diet that was nutritionally balanced and provided 35% of calories as ethanol. Control animals were pair-fed and received the same liquid diet with maltose-dextrins substituting for ethanol. At birth the pups of both the experimental and control groups were fostered by surrogate mothers which received normal rat chow ad libitum. An average of eight alcohol-fed and eight pair-fed pups were killed at 0–1, 5, 7, 10, 14, 21, 28, and 42 postnatal days. The fixed cerebella of these pups was bisected in the midvermal plane and one-half was embedded in paraffin and the other in Araldite. Projection drawings of hematoxylin and eosin-stained paraffin sections were made at 50× magnification. Using the graphics plate of an Apple II computer, the circumference and area of the vermal cerebellum was determined. We found that at all the time sequences studied the area and circumference of the cerebellum were significantly reduced in ethanol-exposed pups compared with the pair-fed controls.

A strain difference in the adrenal zona glomerulosa determined by one gene-locus.

The zona glomerulosa is well developed in the adrenal glands of CBA/FaCam mice and poorly developed in those of Peru mice. Measurements were made of the width of the subcapsular zone in hematoxylin and eosin stained paraffin sections of the adrenals of young adult malemice from these 2 strains, their F1, and the backcrosses to the parental strains.The pattern of results found in these stocks implies that the major portion of the strain difference is controlled by genetic variation at a single locus. The results of progeny tests of individual backcross backcross mice are in agreement with this hypothesis. It is suggested that the dominant allele found in CBA mice be called Ezg and that found in Peru mice, ezg. Sodium deficiency results in glomerular hypertrophy in CBA mice but not inPeru mice, even though urinary sodium measurements implied that both kinds of mice could effectively conserve sodium. The relationship between the difference in the zona glomerulosa and known differences in the metabolism of el...