Increased density of inhibitory noradrenergic parenchymal nerve fibers in hypertrophic islets of Langerhans of obese mice.

BACKGROUND AND AIM: We sought to identify mechanisms of beta cell failure in genetically obese mice. Little is known about the role of pancreatic innervation in the progression of beta cell failure. In this work we studied adrenergic innervation, in view of its potent inhibitory effect on insulin secretion. We analyzed genetically obese ob/ob and db/db mice at different ages (6- and 15-week-old), corresponding to different compensatory stages in the course of beta cell dysfunction. 15 week-old HFD mice were also studied. METHODS AND RESULTS: All mice were characterized by measures of plasma glucose, insulin, and HOMA. After perfusion, pancreata were dissected and studied by light microscopy, electron microscopy, and morphometry. Insulin, Tyrosine Hydroxylase-positive fibers and cells and Neuropeptide Y-positive cells were scored by immunohistochemistry. Islets of obese mice showed increased noradrenergic fiber innervation, with significant increases of synaptoid structures contacting beta cells compared to controls. Noradrenergic innervation of the endocrine area in obese db/db mice tended to increase with age, as diabetes progressed. In ob/ob mice, we also detected an age-dependent trend toward increased noradrenergic innervation that, unlike in db/db mice, was unrelated to glucose levels. We also observed a progressive increase in Neuropeptide Y-immunoreactive elements localized to the islet core. CONCLUSIONS: Our data show increased numbers of sympathetic nerve fibers with a potential to convey inhibitory signals on insulin secretion in pancreatic islets of genetically obese animals, regardless of their diabetic state. The findings suggest an alternative interpretation of the pathogenesis of beta cell failure, as well as novel strategies to reverse abnormalities in insulin secretion.

HSP70 is selectively over-expressed in the blast cells of the germinal centres and paracortex in reactive lymph nodes.

AIMS: We evaluated by immunohistochemistry HSP70 expression in reactive lymph nodes since its morphological expression and location have not been previously described and correlated with lymphocyte kinetics. METHODS AND RESULTS: Ninety-six cases of non-specific lymphadenitis were immunostained for HSP70, CD20, CD3, Ki67, Bcl-2, CD21. The type and the location of HSP70-positive cells were determined. Their number out of 2000 cells in each germinal centre and in each paracortical area was counted at 60x magnification with the help of a quantitative grid. Seventeen percent of germinal centre cells and 7.6% of the paracortex cells were positive. This difference was highly significant. The positively reacting cells were B-cells and had a blast (centroblast or immunoblast) morphology, with negative mantle and marginal lymphocytes and T-cells. Lymphoplasmacytoid cells and plasma cells reacted only weakly or were negative. Germinal centre antigen-presenting cells and interdigitating dendritic cells reacted from lightly to moderately. CONCLUSIONS: HSP70 was selectively over-expressed by B-blasts mainly located within germinal centres with a lower number in the paracortex. The difference in the mean number between the two sites was statistically highly significant. No correlation was found with bcl-2 and Ki67 expression. Mantle, marginal and T-lymphocytes were always negative. The biological meaning and role of this over-expression in centroblasts and immunoblasts remain to be elucidated.

Serum markers for monitoring of prostatic carcinoma.

BACKGROUND: Serum TPS (tissue polypeptide-specific antigen) has been observed to be characteristic of carcinoma proliferation, and increased levels of TPS seem to be closely related to tumor progression. In this study we wanted to evaluate the importance of the tumor-marker TPS in the diagnosis and follow-up of patients with prostatic carcinoma, and to compare it with prostate-specific antigen (PSA). METHODS: We considered 39 patients with clinically confined disease, who underwent neoadjuvant hormonal therapy and thereafter radical prostatectomy, and 45 patients who did not undergo surgery and underwent hormonal adjuvant therapy alone. PSA and TPS were measured at the time of diagnosis and at regular intervals in the follow-up; TPS was measured in a control group of patients as well. RESULTS: We were able to observe that, in untreated patients, PSA correlates with clinical stage, increasing with increasing tumor stage; a similar correlation was not observed when considering TPS. After androgen ablation we observed a decrease in PSA, but the serum values of TPS remained higher, suggesting that activity still exists inside the tumor. The evaluation of TPS appeared to be of particular interest in the follow-up after radical prostatectomy, especially in patients undergoing hormonal therapy; in fact, we were able to observe that relapse of the disease can be suspected early by the increase of TPS in hormonally treated patients. CONCLUSIONS: We assert that TPS can add useful information on the state of neoplastic illness, especially in patients following adjuvant androgen-suppressive hormonal therapy, after radical prostatectomy; serial measurements of this marker could be useful in the early diagnosis of a relapse.

[Gene p53 mutation and prognosis in bladder tumors]. / Mutazione del gene p53 e prognosi nei tumori vescicali.

The choice of treatment for bladder tumors is based on pathological criteria, i.e. staging and grading mainly. It has recently been observed that genetic alterations that involve activation of oncogenes and inactivation of suppressor genes can occur during tumorigenesis. p53 protein is coded for by a genes on chromosome 17, and is able to suppress malignant transformation and proliferation; it can be important in maintaining the integrity of DNA, when damaged during neoplastic disease. We have analyzed samples of transitional cell carcinoma of the bladder of 60 patients managed by conservative treatment; all the specimens have been stained for p53 protein. We could observe that the presence of the protein p53 is correlated with staging and grading and is associated with an increased risk of relapse and progression. Patients with transitional-cell carcinoma confined to the bladder that demonstrates nuclear p53 reactivity should be considered for protocols of adjuvant treatment.

Renal oncocytoma. Pathological evaluation and clinical implications.

Renal oncocytoma is a benign tumour of renal tubular origin; oncocytes are transformed epithelial cells rich in mitochondria, probably representing senescent degenerative cellular changes. Most of renal oncocytomas usually follow a benign clinical course and partial nephrectomy or enucleation has been advocated as curative. By immunohistological staining of tissue sections using monoclonal antibodies (DBA, SBA, PNA, UEA, Cytocheratine), we can suppose the histogenetic origin of renal oncocytomas from a region other than the proximal tubular epithelium, and in particular from the collecting duct epithelium. We believe that it is most important to perform flow cytometry to study the chromosomal pattern of the tumour, once intra-operative frozen sections have advanced the suspicion of renal oncocytoma; if oncocytic cells show a diploid pattern, and the tumour mass is well circumscribed and has not an excessive diameter, we favour renal sparing surgery.

Extraskeletal osteosarcoma of the mediastinum associated with long-term patient survival. A case report.

The authors report a case of mediastinal extraskeletal osteosarcoma showing immunohistochemical and ultrastructural features of epithelial differentiation and associated with a long-term patient survival. The possible role of flow cytometric DNA analysis in defining the prognosis of this tumor is discussed.

Prostatic invasive adenocarcinoma. Effect of combination endocrine therapy (LHRH agonist and flutamide) on the expression and location of proliferating cell nuclear antigen (PCNA).

Expression and location of Proliferating Cell Nuclear Antigen immunostaining in epithelial nuclei were assessed on histological sections from 32 cases of invasive adenocarcinoma of the prostate gland: 20 untreated and 12 treated with combination endocrine therapy or CET. The PCNA-positive nuclei showed homogeneous or granular types of staining or a mixture of both, and a gradation in the intensity of staining. Nuclei with homogeneous patterns appeared darker brown than the lighter granular and mixed patterns. Darker nuclei were more frequently noted, mainly among the epithelial cells adjacent to the stroma, in the untreated cases. In contrast, nuclei with pyknotic chromatin, unstained and corresponding to apoptotic bodies, were more frequently seen in the treated patients. For the untreated invasive adenocarcinomas, the mean proportion of PCNA-stained epithelial nuclei in the 10 cases with an acinar pattern (small and large) was 8.86% (SE 0.23%). The mean value in the 5 cases with a cribriform pattern was 11.76% (SE 0.52%), that is, greater than in the acinar pattern, and decreased from the nuclei in the basal position, or adjacent to the stroma, toward the lumen: 14.40% (SE 0.61%) in the basal position, 11.84% (SE 1.30%) in the intermediate and 9.26% (SE 0.66%) in the lumenal. In the 5 cases with a solid/trabecular pattern, the proportion of PCNA-positive nuclei was 15.74% (SE 2.30%), that is, higher than in all the other patterns, and decreased from the cell layer adjacent to the stroma (17.60%, SE 2.92%) toward the other layers (13.88%, SE 1.71%).(ABSTRACT TRUNCATED AT 250 WORDS)

Frequency and location of mitoses in prostatic intraepithelial neoplasia (PIN).

The aim of our study was to assess the frequency and location of mitoses in routine haematoxylin- and eosin-stained sections of prostatic intraepithelial neoplasia (PIN) and then to compare the patterns with those in benign prostatic hyperplasia (BPH) and prostatic invasive adenocarcinoma (PAC). The frequency of mitoses in the epithelial cell layers increased from BPH through PIN up to PAC. The proportions of mitoses in PIN lesions of low grade (PINlow) and high grade (PINhigh) were greater than in BPH (mean 0.001%; standard error, SE, 0.001%), the values decreasing from the basal layer towards the lumenal. In PINlow, the mean category values were 0.087% (SE 0.04%) in the basal, 0.046% (SE 0.033%) in the intermediate and 0.024% (SE 0.024%) in the lumenal position. In PINhigh, the mean category values were 0.194% (SE 0.178%) in the basal position, 0.075% (SE 0.06%) in the intermediate and 0.049% (SE 0.033%) in the lumenal position. The proportions of mitoses in adenocarcinoma with cribriform pattern decreased from the basal towards the lumenal layer, as for PIN: 0.154% (SE 0.096%) in the basal position, 0.072% (SE 0.044%) in the intermediate and 0.064% (SE 0.04%) in the lumenal position. In the solid/trabecular adenocarcinomas, the mean category value in the cell layer adjacent to the stroma was 0.22% (SE 0.111%), whereas in the other cell layers it was 0.074% (SE 0.045). In small and large acinar adenocarcinomas, the proportions of mitoses were 0.058% (SE 0.024%) and 0.068% (SE 0.019%), respectively. In conclusion, the evaluation of mitotic frequency and location in haematoxylin- and eosin-stained sections gives accurate information on how the mitotic activity in PIN compares with BPH and PAC.

Computed cell cycle and DNA histogram analyses in image cytometry in breast cancer.

AIMS: To analyse the cell cycle and DNA histogram components in data from DNA static cytometry and, in particular, to investigate the influence of the length of time the slides are exposed to the light of the cytophotometer in evaluating the G0/G1 peak. METHODS: DNA static cytometry was performed on 18 Feulgen stained imprints and six histological sections taken from six breast carcinomas. The total optical density values obtained were analysed using software commercially available as Multicycle. DNA flow cytometry was performed on the same cases. RESULTS: The proportions of nuclei related to the cell cycle components from DNA static cytometric data, obtained from Feulgen stained cytological smears, were almost identical with those obtained from DNA flow cytometric data. Moreover, additional information was obtained from the DNA static cytometry frequency histogram and the proportions of nuclei below the diploid G0/G1 peak and above the G2 phase. Discrepancies between DNA static cytometry and DNA flow cytometry were seen in the large coefficients of variation of the G0/G1 peaks obtained with the former method of analysis, even though a better correspondence was found when the exposure time of the slides to the light of the cytophometer was conspicuously shortened. The information obtained from histological sections seemed to be similar to that obtained from DNA flow cytometry when a single cell population was present; a single cell population was detected in two out of the three cases in which two distinct populations had been present in DNA flow cytometry. CONCLUSIONS: The computer analysis of DNA static cytometric data obtained from Feulgen stained cytological specimens provides the type of information on the cell cycle which is usually obtainable only from DNA flow cytometry. Correspondence with the DNA data from histological sections, however, was poor.

Proliferating cell nuclear antigen (PCNA) evaluation in the diagnostic quantitative pathology of cribriform adenocarcinoma of the prostate.

The aim of this study was to emphasise the importance of the measurement sites in tumours from the biological point of view. In particular, two distinct aspects regarding locations were investigated for Proliferating Cell Nuclear Antigen (PCNA) expression in the cribriform adenocarcinoma of the prostate as an example. The first aspect was the identification of the most suitable part of the tumour nodule to be analysed, that is, periphery or marginal zone vs central. The second aspect consisted of the precise location of the objects in relation to the histologic pattern and its components, such as the different cell layers in the cribriform pattern. The results obtained showed that the proportion of PCNA-immunostained nuclei in the marginal zone of the tumour decreased from the basal position, or adjacent to the stroma, towards the lumen: 14.40% (standard error, SE, 0.61%) in the basal position, 11.84% (SE 1.30%) in the intermediate and 9.26% (SE 0.66%) in the lumenal position. In the central zone of the tumour the trend of value changes was similar to that obtained in the marginal zone. However, the proportions were lower and the differences statistically significant. In conclusion, the degree of PCNA expression is related to both locations. Thus, adequate information on the biology of the lesions can only be obtained when the precise site of the objects to be evaluated is identified. Otherwise, misleading results about the lesions being measured can be derived.

Prostatic intra-epithelial neoplasia: expression and location of proliferating cell nuclear antigen in epithelial, endothelial and stromal nuclei.

The expression and location of proliferating cell nuclear antigen (PCNA) immunostaining in epithelial, endothelial and stromal nuclei were assessed in prostatic intra-epithelial neoplasia (PIN). It was then compared with patterns in benign lesions and in invasive adenocarcinomas of the prostate. The PCNA-positive nuclei showed homogeneous or granular types of staining, or a mixture of both, and a gradation in the intensity of staining. Nuclei with granular and mixed patterns appeared lighter brown than those with a homogeneous pattern, which are darker and more often noted in PIN and invasive adenocarcinomas than in benign lesions. For epithelial PCNA-stained nuclei, the proportions in the two grades of PIN were greater than in benign prostatic hyperplasia (mean 3.16%, SE 0.31%) and prostatic atrophic ducts and acini (mean 0.56%, SE 0.09%), the values decreasing from the nuclei in the basal position towards those in the luminal layer. In grade 1, the category mean values were 9.51% (SE 1.14%) in the basal, 7.02% (SE 1.27%) in the intermediate and 6.02% (SE 0.90%) in the luminal position. In grade 2, the category mean values were 13.81% (SE 1.42%) in the basal position, 10.99% (SE 1.17%) in the intermediate and 7.91% (SE 1.43%) in the luminal position. In small and large acinar adenocarcinomas, the proportions of positive nuclei were 8.66% (SE 0.30%) and 9.06% (SE 0.30%), respectively. The category mean values in the cribriform adenocarcinomas were 14.40% (SE 0.61%) in the basal position, 11.84% (SE 1.30%) in the intermediate and 9.26% (SE 0.66%) in the luminal position. As in PIN, the proportions of immunostained nuclei in the adenocarcinoma with cribriform pattern decreased from the basal towards the luminal layer. In the solid/trabecular adenocarcinomas, the category mean value in the cell layer adjacent to the stroma was 17.60% (SE 2.92%), whereas in the other cell layers it was lower than that in the cells adjacent the stroma (mean 13.88%, SE 1.71%). For capillary endothelial and stromal cells, the percentages of PCNA-stained nuclei were much lower than those in the epithelial component. The lowest mean values were obtained in benign lesions, whereas the highest were in invasive adenocarcinomas, the percentages in PIN being intermediate.

Proliferating cell nuclear antigen (PCNA) in prostatic invasive adenocarcinoma. Is the proliferation state in the marginal zone of the tumour higher than in the central part?

Expression and location of Proliferating Cell Nuclear Antigen in epithelial nuclei were assessed in invasive adenocarcinoma of the prostate gland. The PCNA-positive nuclei showed homogeneous or granular types of immunostaining or a mixture of both, and a gradation in the intensity of staining. Nuclei with homogeneous pattern appeared darker brown than the lighter granular and mixed patterns. Darker nuclei were quite frequently noted, mainly among the epithelial cells adjacent to the stroma. For the marginal zone of invasive adenocarcinoma, the mean proportion of PCNA-stained nuclei in the small acinar pattern was somewhat similar to that in the large acinar pattern, i.e., 8.66% and 9.06%, respectively. In contrast, the mean values in the cribriform pattern were greater than in the small and large acinar patterns, and decreased from the nuclei in the basal position, or adjacent to the stroma, toward the lumen: 14.40% in the basal position, 11.84% in the intermediate and 9.26% in the lumenal. In the solid/trabecular pattern, the proportions of PCNA-positive nuclei were higher than in all the other patterns: 17.60% in the cell layer adjacent to the stroma and 13.88% in the other layers. The trend of value changes in the central zone of the tumour was similar to that obtained in the marginal zone. However, the proportions were lower and the differences statistically significant. This might indicate that the proliferation state is higher in the marginal zone and that the tumour grows eccentrically rather than centrally.

Occurrence of cell death (apoptosis) in prostatic intra-epithelial neoplasia.

The aim of our study was to assess the frequency and location of apoptotic bodies (ABs) in haematoxylin and eosin-stained sections of prostatic intraepithelial neoplasia (PIN) and then to compare the patterns with those in benign prostatic hyperplasia (BPH) and prostatic invasive adenocarcinoma (PAC). ABs were identified in all epithelial cell layers of the ducts, acini and tumour islands, as well as in the lumina contained in such structures. In the epithelial cell layers, ABs were found in general in the intercellular space and occasionally in the cytoplasm of epithelial cells. The frequency of ABs increased from BPH through PIN up to PAC. The proportions of ABs in PIN lesions of low grade (PINlow) and high grade (PINhigh) were greater than in BPH, the values decreasing from the nuclei in the basal position towards those in the luminal layer. In PINlow, the mean category values were 0.85% (standard error, SE, 0.311%) in the basal, 0.623% (SE 0.065%) in the intermediate and 0.474% (SE 0.138%) in the luminal position. In PINhigh, the mean category values were 1.006% (SE 0.16%) in the basal position, 0.713% (SE 0.182%) in the intermediate and 0.618% (SE 0.172%) in the luminal position. The proportions of ABs in adenocarcinoma with cribriform pattern decreased from the basal towards the luminal layer, as for PIN: 1.806% (SE 0.346%) in the basal position, 1.15% (SE 0.172%) in the intermediate and 0.886% (SE 0.137%) in the luminal position. In the solid/trabecular adenocarcinomas, the mean category value in the cell layer adjacent to the stroma was 2.154% (SE 0.203%), whereas in the other cell layers it was 2.052% (SE 0.239%). In small and large acinar adenocarcinomas, the proportions of positive nuclei were 1.022% (SE 0.1%) and 0.922% (SE 0.163%), respectively. The evaluation of the frequency and location of ABs gives accurate information on cell death in PIN in comparison with BPH and PAC.