Differential histochemical peanut agglutinin stain in benign and malignant human prostate tumors: Relationship with prostatic specific antigen immunostain and nuclear DNA content

The histochemical binding pattern of the peanut (Arachis hypogaea) lectin (PNA) was quantitatively described by means of computer-assisted microscope analysis in 28 benign prostatic hyperplasias (BPH), 15 prostatic intraepithelial neoplasias (PIN), and 119 prostatic adenocarcinomas. PNA exhibits noninunune but selective binding to glycoproteins with β-D-galactosyl(1,3)-N-acetyl-D-galactosamine residues. We also investigated whether a relationship existed between the number of histochemical-related PNA acceptors and the histochemical prostate-specific antigen (PSA) stain intensity, and between the number of PNA receptors and DNA ploidy level. The results show that neoplastic prostate tissues and high-grade intraepithelial prostatic neoplasias (PIN2_3) exhibit a significantly higher number of PNA acceptors than benign prostatic hyperplasias and low (PIN1) grade prostatic intraepithelial neoplasias. A statistically significant correlation was observed between the number of histochemically related PNA acceptors and PSA immunostain intensity. Lastly, diploid prostatic tumors, whether benign or malignant, exhibited a significantly higher number of PNA acceptors than aneuploid ones. These results suggest that PNA acceptors play an important role in the biology of prostate tumors.     

DNA histogram typing and DNA index measurements in 508 invasive breast carcinomas from fine-needle aspirates and imprint smears as opposed to formalin-fixed paraffin-embedded tissues

Morphometric, i.e. nuclear area (NA), and densitometric, i.e. nuclear DNA content, features were characterized in a series of 508 invasive ductal breast carcinomas. The specimens analyzed were from three distinct sources, i.e. fresh material (252 fine-needle aspirates as opposed to 147 imprint smears) immediately fixed in EFA fixative as opposed to archive material, i.e. 109 formalin-fixed paraffin-embedded tumours that were subsequently deparaffinized. Morphonuclear parameters were computed on Feulgen-stained nuclei by means of a cell image processor. Our results show that the development of nuclear size and DNA content in function of anatomopathological grading is approximately the same for specimens of breast cancer provided by FNAs, imprint smears and formalin-fixed paraffin-embedded tissues. However, in these latter instances it seems that part of the morphometric information is slightly modified in relation to the information obtained from fresh material directly fixed in EFA for cytophotometric analysis. The greatest discriminatory power of the morphometric parameters was obtained in relation to the FNAs. Lastly, in the present study we come out in favor of the idea that henceforth it would be preferable to express results concerning nuclear DNA content as DNA histogram types rather than in terms of DNA indices.     

In-vitro characterization of dihydrotestosterone-induced, epidermal growth factor-induced and basic fibroblastic growth factor-induced modifications in the growth dynamics of the human prostate-cancer cell-line lncap, du145 and pc3

The influence of dihydrotestosterone (DHT), the epithelial growth factor (EGF) and the basic fibroblast growth factor (bFGF) was investigated on LNCaP, DU145 and PC3 cell growth, which represents the ratio between cell gain (cell proliferation) and cell loss (cell death). In the present study, cell growth was assessed by means of the computer-assisted microscope analysis of Feulgen-stained nuclei combined with the mathematical Delaunay triangulation and Voronoi paving techniques, which enabled the cell colony patterns, i.e. their density and level of organisation, to be determined. The results from a previous study (Janssen et al, Prostate, in press) combined with those of the present one show that DHT was found to activate proliferation of the LNCaP model, as evidenced by increase in size of colonies, increase in number of cells within colonies, increase in cell colony density and, accordingly, decrease in mean segment length value (which is the distance between adjacent cell nuclei). Using the same criteria, DHT was found inhibitory on growth of DU145 cell line, and devoid of significant effect on PC3 cell line. Basic FGF was found to be a powerful stimulator of growth of PC3 cell Line and to induce a weaker stimulation of DU145 cell line. On LNCaP cell line, it increased the size of colonies without increase of the number of cells per colony. This feature can be explained by a decrease in cell colony density. With respect to the same colonies, the proliferation index (percentage of cells in the S+G2 phases of the cell cycle) was found similar to that of the controls. This suggests that the increase in the size of the colonies is due to a difference of spreading of the cells on their supports. EGF had no significant effect on LNCaP and PC3 models, and was decreasing cell density of DU145 colonies.     

The characterization of biological features in dedifferentiated liposarcomas by means of principal components and discriminant analyses of 25 computer-generated variables from Feulgen-stained nuclei and histological slides

Several groups of lipomatous tumors are not yet clearly characterized on the biological level. In order to attempt to classify the dedifferentiated liposarcomas with respect to other types of malignant liposarcomas, 80 adipose tumors were submitted to the combination of two computer-assisted methodologies. These two methodologies consisted of i) the determination of 25 variables, and ii) the analysis of the diagnostic information contributed by these 25 variables by means of two complementary techniques, i.e. principal components and discriminant analyses. The 25 variables were computed by means of image cytometry on Feulgen-stained nuclei and histological slides, quantitatively describing distinct biological characteristics relating to morphonuclear (chromatin pattern) features (14 variables), nuclear DNA content distribution (9 variables), and tissue architecture pattern (2 variables). The 80 adipose tumors included 21 typical lipomas, 7 atypical lipomas (defined as extremity adipose tumors with a histopathological pattern of well-differentiated liposarcomas), 16 retroperitoneal and 5 non-retroperitoneal abdominal well-differentiated liposarcomas, 9 dedifferentiated liposarcomas, 8 myxoid (intermediate-grade tumor) and 14 pleomorphic (high-grade tumor) liposarcomas. The data strongly suggest that the dedifferentiated liposarcomas exhibit biological characteristics which are distinct from those of low- and high-grade liposarcomas, but similar to those of intermediate ones. The results also show that typical and atypical lipomas are two distinct biological entities. In contrast, the atypical lipomas and the well-differentiated retroperitoneal and non-retroperitoneal liposarcomas exhibited a high number of similar biological characteristics. Computer-assisted methods contribute valuable information to characterize lipomatous tumor biology.     

Characterization of chemotherapy-induced morphonuclear modifications in the P388 leukaemia and the MXT mammary tumour models of the mouse

Chemotherapy-induced morphonuclear modifications were monitored in vivo by means of the digital cell image analysis of Feulgen-stained nuclei. Two experimental models were used, i.e. the P388 mouse leukaemia and the MXT mouse mammary carcinoma. The drugs used were doxorubicin, etoposide and cyclophosphamide. The results indicate that the chemotherrapy induced a significant decrease in the MXT tumour growth and a significant increase in the survival of the P388 leukaemic mice. These effects were accompanied at the morphonuclear level by an increase in the nuclear area, by modifications in the DNA content in accordance with the effects of the drugs on the cells cycle and by several modifications in the chromatin texture in accordance with the effects of the drugs on the cells cycle and by several modifications in the chromatin texture in accordance with the model or drugs studied. While there were neither homogeneous morphonuclear changes in all treatment groups nor clearcut correlations between the morphonuclear changes and tumour growth or the survival of the animals, the present study nertherless shows that it is possible, at least partly, to monitor in vivo certain chemotherapy-induced effects occurring at the morphonuclear level, and subsequently to obtain information on the mode of action of the drugs.     

Triterpene saponins, quaternary ammonium compounds, phosphatidyl cholines, and amino acids in the pronotal and elytral secretions of Platyphora opima and Desmogramma subtropica

Secretions of the pronotal and elytral glands of adults of the chrysomelid beetle Platyphora opima from Panama have been shown to contain two oleanane triterpene saponins: the known 3-O-beta-D-glucopyranosyl-(1-->3)-beta-D-glucuronopyranosyl-oleano lic acid-28-O-beta-D-glucopyranoside and compound 1, whose structure was established as 3-O-beta-D-glucopyranosyl-(1-->3)-beta-D-glucuronopyranosyl-29- hydrox yoleanolic acid-28-O-beta-D-glucopyranoside by a combination of 1D and 2D NMR methods (COSY, HMQC, HMBC, and TOCSY) and FABMS. The secretions also contained N,N,N-trimethylcadaverine and its 1, 2-dehydro derivative 3, as well as the nicotinamide derivative 4. Secretions of Desmogramma subtropica, also from Panama, contained as sole triterpene derivative 3-O-beta-D-glucopyranosyl-(1-->2)-beta-D-glucuronopyranosyl-24- hydrox yoleanolic acid (2), together with glutamic acid, glutamine, pyroglutamic acid, and arginine. A mixture of phosphatidylcholines was also present in the secretions of both species.