Evaluation of pathological parameters and morphometric data of desmoplastic lobular breast carcinoma.

Background: Invasive lobular breast cancer (ILC) is the second most frequent form of breast cancer. While cancer cells are regularly investigated, tumor stroma represents a highly unexplored field. Aims: The aim of this study is to perform a detailed investigation of clinical, immunohistochemical, and morphometric characteristics of desmoplastic (D) and nondesmoplastic (ND) ILC. Materials and Methods: This study included twenty cases of ILC that were divided into two groups designated as D and ND groups. Medical histories and diagnosis data were obtained from the archives of the Center of Pathology, Clinical center Niš (Serbia). Morphometric analysis of hematoxylin and eosin stained slides was performed using ImageJ software, and the obtained data were further statistical processed. Results: Statistical analyses of the data revealed that no significant differences between D and ND groups when patient age, estrogen receptor (ER), and progesterone receptor (PR) expressions and morphometrical parameters (such as the distance between groups of cancer cells and nucleocytoplasmic ratio) were compared. However, D and ND groups statistically, significantly differed in the occurrence of axillary lymph node metastasis, and when the ER and PR data were included, in certain nuclear parameters (cell/nucleus area, perimeter, Feret’s diameter, and circularity). Conclusions: Desmoplastic stroma was observed more frequently in patients without axillary lymph node metastases, whereas the expression of ER and PR had no influence on its development. According to the measured morphometric parameters larger cells/nuclei belonged to ND group.

Image analytic study of nuclear area in mantle cell lymphoma

BACKGROUND: Malignant lymphomas are classified on the basis of morphology, immunohistochemistry, and genetic and molecular biological features. Morphology is considered the most important and basic feature. Lymphomas can be classified as small, medium, or large depending on the cell size, but this criterion tends to be rather subjective. The aim of this study was to investigate the usefulness of an objective approach based on quantitative measurements. METHODS: Twenty specimens of mantle cell lymphoma and 2 specimens of the tonsil were examined. The nuclear area of 6,401 tumor cells of mantle cell lymphoma and 743 normal mantle cells of reactive tonsils were measured by 3 authors by using a user-controlled image-analyzer. The images of the nuclei were outlined using the spline method and the i-solution software, and the data were assessed using ANOVA and Student's t-test. RESULTS: The mean nuclear areas of mantle cell lymphoma cells measured by the 3 authors were 37.9 [7.9] microm2, 37.9 [7.2] microm2, and 38.2 [7.7] microm2 and those of normal mantle cells in reactive tonsil were 28.6 [2.3] microm2, 28.8 [2.0] microm2, and 27.0 [3.0] microm2. There was no statistical difference between the 3 observations of mantle cell lymphoma (P=0.580) and normal tonsils. CONCLUSION: For morphology, nuclear area is considered an important feature in the classification schemes of lymphoma. We showed that nuclear area measurement by using image analyzer can be used as an objective quantitative method. We think that nuclear morphometry may play a significant role in the diagnosis of lymphoma.

Argyrophilic Nucleolar Organizer Region and Expression of Ki-67 in Malignant Lymphoma

Nucleolar organizer regions (NORs) are loops of DNA which occur in the nucleoli of cells which possess ribosomal RNA genes. The numbers and areas of NORs have been thought to be related to cellular activities. We aimed to investigate the direct relationship between the parameters of AgNORs and cellular proliferative activity using immunohistochemical method with the monoclonal antibody Ki-67, which demonstrates proliferating nuclei. The sequential technique for the simultaneous Ki-67 immunostaining and NOR staining was applied to the same slides of a series of non-Hodgkin's lymphomas (NHL) of the low, intermediate, and high grade type. The number of AgNOR per cell was counted and mean NOR percentage nuclear area (NPNA) was measured by morphometry in both the Ki-67 positive and Ki-67 negative nuclei. The increased immunoreactivity for Ki-67 was found in the high grade than in the low grade non-Hodgkin's lymphoma. This was reflected in the two areas of the palatine tonsils and lymph nodes, the positive cell counts being higher in the follicle center nuclei than in those in the interfollicular compartment. In general the numbers and NPNA of AgNORs were higher in the Ki-67 positive nuclei than in those lacking the antigen in malignant lymphomas as well as in control. The AgNORs numbers and NPNA in controls were the highest in the Ki-67 positive cells in the follicular area and the lowest in the Ki-67 negative cells in the perifollicular area. In malignant lymphomas the numbers and NPNA of AgNORs tended to increase in proportion to their grade in both the Ki-67 positive and negative cells. The numbers of AgNORs of the high grade and the NPNA of the intermediate and the high grade were significantly higher in Ki-67 positive cells than in Ki-67 negative ones. With this double staining method it was now possible to confirm that numbers and NPNA of AgNORs were directly related to the cellular proliferative activity. In maligant lymphoma, among the several parameters of the AgNORs, NPNA in Ki-67 positive cells is proposed to be the most useful marker in determining the prognosis of the patient.

Significance of S-phase Fraction and Nuclear Area in Classification and Prognosis of Non-Hodgkin Lymphoma / 대한혈액학회지

BACKGROUND: Non-Hodgkin lymphomas have morphologic and biologic variabilities and are classified by a variety of classification schemes. The authors studied the value of DNA content, S-phase fraction, and nuclear areas as a diagnostic discriminant in classification and predicting the clinical outcome of non-Hodgkin lymphoma. METHODS: Thirty cases of non-Hodgkin lymphoma and 13 cases of reactive lymph node specimens embedded in paraffin were used. DNA analysis was done by flow cytometric method using propidium iodide staining. Nuclear areas of lymphoma cells were measured by image analyzer. The results of DNA content, S-phase fraction, nuclear areas were statistically analyzed between groups of lymphoma classified according to the Rappaport classification and Working formulation. RESULTS: Aneuploidy patterns were identified in two cases (6.7%) of non-Hodgkin lymphoma (diffuse poorly differentiated lymphocytic and well differentiated lymphocytic). S-phase fraction and nuclear areas were high in lymphoblastic, diffuse poorly differentiated, diffuse lymphocytic undifferentiated, and diffuse mixed types (unfavorable prognostic group), and which had statistical significance against the low values of diffuse lymphocytic well differentiated type (favorable prognostic group). The S-phase fraction and nuclear area had significant correlation, so they could represent correlation between high proliferative activity and large nuclear area. High, intermediate, and low grades groups classified by Working formulation revealed statistically significant differences of S-phase fraction and nuclear areas between each group. CONCLUSION: The S-phase fraction and nuclear area measurement could be good diagnostic and prognostic factors in classification and clinical management of non-Hodgkin lymphoma.