Immunohistochemical detection of cell-cycle associated markers on paraffin embedded and formalin fixed needle biopsies of prostate cancer: correlation of p120 protein expression with AgNOR, PCNA/cyclin, Ki-67/MIB1 proliferation-scores and Gleason gradings.

Paraffin embedded and formalin fixed needle biopsies of prostate cancer (PC) were used to immunocytochemically detect the p120 nucleolar protein in relation to the Gleason histological gradings (GHG), the labelling indices of proliferating nuclear immunocytochemical markers (PCNA/Cyclin, Ki-67/MIB1) and the argyrophilic nucleolar region (AgNOR) rate. The twenty-six cases of PC (6 from large histological samples and 20 from needle biopsies) were equally distributed into low (< or = 6) or high (> or = 7) GHG groups. The p120 nucleolar protein immunocytochemical reaction was randomly expressed in large histological sections but uniformly distributed without gaps in needle biopsy sections. Only on the latter were quantitative values of PCNA/Cyclin (23.2 in low and 45.3 in high GHG), Ki-67/MIB1 (13.8 in low and 43.3 in high GHG) and AgNOR (5.0 in low and 7.5 in high GHG) related to those of p120 nucleolar protein (0.8 in low and 3.8 in high GHG). The values of all these cell cycle markers increased from low to high GHG of PC, all four reaching high statistical significance between the two groups (ANOVA-two tailed p < 0.0001). The PCNA/Cyclin index showed a higher positivity than the Ki-67/MIB1 index in PC with low GHG but not in PC with high GHG. In conclusion, paraffin embedded and formalin fixed PC needle biopsies exhibit a higher diagnostic PCNA/Cyclin than Ki-67/MIB1 index for cases presenting differentiated features, whereas p120 nucleolar protein detection seems to be a suitable marker of poorer outcome of PC.

Intrahepatic bile duct loss in primary sclerosing cholangitis: a quantitative study.

AIMS: Intrahepatic bile duct volume density has been evaluated in four cases of primary sclerosing cholangitis (PSC) by applying a semiautomatic image analysis system to histological sections of surgical biopsies. Comparison was made between normal livers and cases of primary biliary cirrhosis. METHODS AND RESULTS: A marked decrease in both bile duct volume density in liver (up to 50% of normal value) and bile duct volume density in portal tracts (up to 21% of normal value) was found. The analysis of correlations between the portal tract size and the portal tract volume fraction constituted by bile ducts confirms that the destruction mainly affects small and medium-sized ducts. The ratio of bile duct to arterial component volume fractions in portal tracts turned out to be inverted with respect to that determined in normal liver, showing a decrease of up to 30% of normal value. CONCLUSIONS: The features observed in primary sclerosing cholangitis, as well as the extent of bile duct loss proved to be quite similar to what we had previously described in primary biliary cirrhosis. Thus, regardless of the causes and mechanisms leading to the bile duct loss, the quantification of ductopenia produced results which are alike for the two distinct cholestatic diseases.

Quantitative analysis of intrahepatic bile duct component in normal adult human liver and in primary biliary cirrhosis.

The volume density of bile ducts in adult normal liver has been analyzed in order to provide stereometric parameters to which refer on performing quantitative evaluations of bile duct loss in acquired liver diseases. Five livers were studied by applying a semi-automatic image analysis system (ASM 68K Leitz) to histological sections. No significant differences (P > 0.10) were found for all measures and the following values were derived from the pooled data: (1) mean % volume of bile ducts in liver = 0.318 +/- 0.171; (2) mean % volume of portal tracts in liver = 4.351 +/- 2.860; (3) mean % volume of bile ducts in portal tracts = 6.567 +/- 3.813. The parallelism of bile duct to arterial components of portal tracts was also investigated and expressed as the ratio of their respective volume fractions (mean ratio = 1.72). The validity of the obtained parameters was tested by comparing them with values determined in five cases of primary biliary cirrhosis (PBC). All PBC cases showed a marked decrease in both bile duct % volume in liver (ranging between 0.028 and 0.057) and bile duct % volume in portal tracts (ranging between 0.673 and 0.914), as well as inversion of the bile duct to artery volume ratio in portal tracts (ranging between 0.246 and 0.437).

Age-related distribution of endocrine cells in the human prostate: a quantitative study.

A morphometric analysis was performed to obtain quantitative data on age-related changes in prostatic endocrine cell (PrEC) density. Sixty prostates from subjects aged 14-74 years were studied with a semi-automatic image analysis system (ASM 68K, Leitz) applied to sections immunostained for chromogranin A-reactive cells. The highest density of PrECs (0.366 cells/mm of epithelial length) was found in the 25-54 year age group, which was significantly different from that found in prostates of the younger (0.311 cells/mm) and the older (0.261 cells/mm) age groups. The data probably reflect the higher incidence of incompletely developed glandular units in the younger group and the formation of new alveoli related to the usual glandular hyperplasia that occurs with increasing age in the older group.

Effect of vitamin A on chemotactic and chemoinvasive behaviour of an osteosarcoma cell line.

Vitamin A is known to be able to modulate cell growth and differentiation and to act as an inhibitor of the process of carcinogenesis in some experimental models. Here we have studied the effect of different concentrations of vitamin A on chemotactic and chemoinvasive behaviour of a metastatic osteosarcoma cell line. The cell proliferation was partially inhibited in the presence of 10(-5) M retinol after 4 days of incubation. Retinol effect on chemotactic and chemoinvasive activity of osteosarcoma cells seemed to be dose-dependent. The highest retinol concentration used (10(-5) M) had an inhibitory effect on migratory and invasive cell response. Lower retinol concentrations seemed to be able to enhance (10(-8) M) both chemotactic and chemoinvasive activity of osteosarcoma cells. Chemotaxis and chemoinvasion assays provide rapid and quantitative tools to study the "in vitro" behaviour of metastatic cells. Furthermore, they represent a mean to screen for drugs, hormones and other substances able to alter the metastatic phenotype.

[The subfornical organ today: morphological aspects and functional role]. / L'organo subfornicale oggi: aspetti morfologici e ruolo funzionale.

The recognition of the role played by the subfornical organ (SFO) in the central regulation of body water balance has recently aroused new interest in this anatomical formation which remained ignored for a long time. The SFO is included in the group of the circumventricular organs. In higher vertebrates it is adherent to the ventral surface of the fornix and protrudes into the third ventricle at the level of the interventricular foramina, partially covered by the choroid plexus. The SFO appears as a small nodule, rounded or ovoidal in shape, consisting of highly vascularized nervous tissue and lined by ependyma at the ventricular surface. Its structural organization is fundamentally constant and presents only minor differences in the various species. The SFO neuronal perikarya show different aspects which have been classified in four types. However, it is not yet clearly defined if such aspects refer to distinct cell types or to different transitional features. Nerve and glial cell processes form a dense plexus through the SFO and the subependymal area, as well as in the connective tissue perivascular spaces. These may be narrow or wide and surround fenestrated and non-fenestrated capillaries, assuming sometimes a labyrinthine aspect. The ependymal lining of the SFO ventricular surface shows large variations and regional differences concerning the cell height, the number and development of microvilli, the cilia distribution. The structural properties of SFO, which is characterized by a rich and highly permeable capillary bed, by a wide surface area of contact and exchange with the cerebrospinal fluid, by direct and indirect neural connections with a number of regulatory structures, have been considered as the basis for the role of neurohumoral integration that SFO plays in regulating physiological and behavioral responses to water-mineral changes. Much experimental evidence substantiates this function. However, the studies on SFO are increasingly enriching the literature with new experimental, especially physiological and cytochemical, data which may suggest for this organ connections even more extensive and functions even more complex than those until now ascertained.

Histometry of splenic microvascular architecture in hairy cell leukaemia.

In this study we investigated the splenic microvascular architecture in hairy cell leukaemia, in order to provide a morphological basis for the haemodynamic modifications occurring in the disease. When compared with controls, the four leukaemic spleens examined showed a set of changes involving both the arterial and the venous system. A real increase in the absolute volume, surface and length of pulp arterial vessels was present. This increase was not so great as the enlargement of the spleen, thus resulting in a reduced density of distribution of arterial vessels in the infiltrated pulp. Enlargement of pulp cords and sinuses was also present: the pulp cord enlargement was apparent in the unit volume, which resulted in a disproportionately higher increase of the absolute volume, compared with that of sinuses. The sinus-cordal rearrangement and, particularly, the increase in the volume of pulp cords may cause a slowing down of blood cell circulation with resultant increased phagocytosis and hypersplenism. Moreover, it is suggested that the changes observed in the arterial bed of the spleen in hairy cell leukaemia involve both a reduced blood supply per unit volume of splenic pulp and a more marked conditioning of blood cells prior to their screening by cordal macrophages.

Histometric analysis of white pulp arterial vessels in congestive splenomegaly.

A quantitative study of white pulp arterial vessels has been carried out in congestive splenomegaly with hypersplenism. Arteries and arterioles with periarterial lymphatic sheath (PALS) in abnormal spleens show an absolute volume and total length higher than in controls. Moreover, they present a luminal diameter significantly lower than normal. These findings demonstrate an increase of the white pulp arterial bed in congestive splenomegaly. Thus, the arteries with PALS participate in the complex vascular rearrangement which supports the blood hyperflow in congestive splenomegaly.

Red pulp arterial terminals in congestive splenomegaly. A morphometric study.

A quantitative analysis of the red pulp arterial terminals (comprising arterioles = penicilli, arterial capillaries and sheathed capillaries) has been carried out in congestive splenomegaly occurring in active chronic hepatitis, hepatic cirrhosis and idiopathic portal hypertension. Comparisons have been made with control spleens surgically removed for accidental trauma. Red pulp volume enlargement appears to be associated with a progressive lengthening of arterial terminals, which present a length density similar to that of normal spleens. Moreover, a different rearrangement of the various components of the arterial tree is present: in particular, the percent and absolute volumes of arterioles and arterial capillaries show a higher increase than those of sheathed capillaries. The present study points out the involvement of arterial terminals in the pathological rearrangement of the red pulp in congestive spleens, so supporting the hypothesis that splenomegaly does not result simply from passive congestion.

Histometric analysis of sheathed capillaries in congestive splenomegaly.

The structural rearrangement of human sheathed capillaries in congestive splenomegaly with hypersplenism has been evaluated by histometric analysis. In abnormal spleens the external diameter and the wall thickness were decreased, while the internal diameter was increased. Moreover, the total length and the absolute number of sheathed capillaries were increased, though not to the same high degree as the volume of the whole red pulp. Thus, in addition to the well-known changes of the sinuses, also the terminal arterial passages appear to participate in the morphological rearrangement of the red pulp in congestive splenomegaly.

[Vascular flow-regulating devices in the human kidney]. / Dispositive vasali regolatori di flusso nel rene umano.

Small arterious and venous vessels of the juxtamedullary cortex of normal human kidney present wall formations regulating the blood flow. The arterious devices appear either as sphincterial rings at the origins of collateral branches of interlobular arteries, or as groups of myoepithelioid cells in the wall of small arteries immediately preceding the afferent arterioles. The venular devices consist of circular muscular thickenings repeated along the course of the vessel.

[The presence of arteriovenous anastomoses in the human kidney]. / Sulla presenza di anastomosi arterovenose nel rene umano.

The microscopic observation of serial sections obtained from normal human kidneys allowed to demonstrate the presence of arteriovenous anastomoses in the juxtamedullary cortex. In particular one section is shown which clearly presents an anastomotic channel along with its connections to both an afferent arteriole and a venule.

Changes in the small biliary passages in the hepatic localization of Hodgkin's disease.

The authors have investigated the behaviour of the small biliary passages in the liver biopsies of six patients suffering from untreated Hodgkin's disease with hepatic localization. No obstruction of the major bile ducts was demonstrated in any patient. Three of the patients were anicteric, while the three others presented with jaundice. In the first three cases typical Hodgkin's granulation tissue appears to be limited to portal tracts and collagen reaction is virtually absent. The three cases with cholestasis showed granulomatous tissue associated with heavy connective tissue rearrangement invading and dissociating the lobular structure. They also show a conspicuous bile-duct proliferation, which is not observed in the three anicteric patients. In these latter cases, however, the small bile ducts running within or near the granulomatous tissue present various morphologic changes, including basal membrane thickening, dilation or constriction of the lumen and alterations of the biliary epithelial lining. Complete disappearance of the bile duct may occur.

Stereometry of the red pulp on the human spleen. I) Methodological remarks.

The principal histometrical formulas to be applied to the analysis of human spleen's red pulp are illustrated. The discussion particularly concerns the formulas to evaluate the following measures per unit splenic volume:volume of the elements; surface area of sinus-splenic cord boundaries; mean breadth of cords; length of the sinus; mean sectional area of the sinus. Suggestions are given about the criteria to be followed in the proper assumption of the data which must be subjected to statistical estimation.

Stereometry of the red pulp on the human spleen. II) The normal spleen.

The stereometric measurements obtained in three human normal spleens surgically removed for trauma have been submitted to statistical evaluation. On the basis of the original counts, some stereometric measures of the normal splenic red pulp have been determined, namely per cent volume of sinuses, per cent volume of cords, breadth of the cords, mean sectional area of sinuses and the volume of sinuses:volume of cords ratio. These data can constitute a series of parameters to which compare the measures of the pathologic spleens.

Cytophotometric characterization of cortical and medullary lymphocytes in the chicken bursa and thymus.

We have evaluated by means of cytophotometric techniques the nuclear content in DNA, in total nucleic acids and in histone proteins and the nuclear volume of the thymic and bursal lymphocytes in adult chickens. We have also made the same determinations in the developing bursal follicles before and after hatching. The results indicate that cortical differ from medullary lymphocytes for all the parameters considered both in thymic lobules and bursal follicles. Furthermore these differences appear analogous in both the organs, independently from the fact that they produce precursors of T and B lymphocytes respectively. As concerns the developing bursal follicle, the lymphocytes show the characteristics of the adult medullary lymphocytes. At the hatching changes occur in the nuclear content of total nucleic acids and histones. This is probably related to the exposure to antigenic stimulation through the cloaca.

Phospholipids bound to acidic nuclear proteins in human B and T lymphocytes.

Chromatin fractions (DNA, histones and non-histone chromosomal proteins NHCP) have been isolated from human peripheral B and T lymphocytes using different methods and analyzed in order to identify their lipid content. While DNA and histone fractions do not reveal the presence of lipids, a 2% of phospholipids is present in the NHCP fraction. The phospholipids associated with NHCP present a constant relative ratio among sphingomyelin, phosphatidyl-choline and phosphatidyl-ethanolamine both in B and T lymphocytes, whichever are the extraction procedures employed. These findings are related to the possible dereprssive role of phospholipids on DNA-dependent RNA synthesis.