Search

Lynch syndrome is caused by germline mutations in any of the MisMatch Repair (MMR) genes. About 37% of MSH2 variants are missense variants causing single amino-acid substitutions. Whether missense variants affect the normal function of MMR proteins is crucial both to provide affected families a more accurate risk assessment and to offer predictive testing to family members. Here we report one family, fulfilling both Amsterdam I and II criteria and Bethesda guidelines, referred to our center for genetic counselling. The proband and some of her relatives have been investigated for microsatellite instability (MSI), immunohistochemical MMR protein staining, direct sequencing and Multiplex Ligation-dependent Probe Amplification (MLPA). Also Subcellular Localization Assay and Splice site predictions analyses were used. A germline missense variant of uncertain significance (exon 3, p.Val161Asp) was found in MSH2 gene in proband and in some relatives. The variant was associated with lack of expression of MSH2 protein (DMMR) and MSI-High status in tumour tissues. The localization assay of the MSH2 protein showed an abnormal subcellular localization pattern of the corresponding protein. Finally, splice-site prediction analysis ruled out a potential role of new splice sites as the cause behind the lack of expression of MSH2 protein and we suppose a potential correlation with other forms of post-transcriptional regulation (circular RNAs). The variant here reported shows a high correlation with phenotype and is located in an evolutionary conserved domain. The localization assay also suggest a potential pathogenic role, thus supporting further research on this matter.

Epithelial-to-mesenchymal transition (EMT) is a developmentally vital reversible process by which fully differentiated cells lose their epithelial features and acquire a migratory mesenchymal phenotype. EMT contributes to the metastatic potential of tumors. The expression profile and other biological properties of EMT suggest potential targets for cancer therapy, including in renal-cell carcinoma (RCC). The preclinical and clinical results have substantiated the promises that dysregulated elements leading to EMT can be a potential target in RCC patients. In this study, we illustrated the pathogenic and prognostic role of EMT in RCC. In addition, we reconstructed, by literature analysis, the different pathways implicated in the EMT process, thus supporting the rational for future EMT-directed therapeutic approaches for RCC patients.

Prostate tumour overexpressed 1, PTOV1, was recently identified as a novel gene and protein during a differential display screening for genes overexpressed in prostate cancer (PCa). It has been suggested that overexpression of PTOV1 can contribute to the proliferative status of prostate tumour cells and thus to their biological behaviour. PTOV1 and Ki67 were immunohistochemically evaluated in PCa, atypical adenomatous hyperplasia (AAH), high-grade prostatic intraepithelial neoplasia (HGPIN), and normal-looking epithelium (NEp) of the transition zone (TZ) in 40 radical prostatectomies with pT2a Gleason score 6 PCa (20 with AAH and 20 with HGPIN) and in 10 simple prostatectomies (SPs) (5 with AAH and 5 with HGPIN). The aim was to evaluate PTOV1 protein expression as a marker for tumor development and progression from AAH to PCa. The proportions of PTOV1 and Ki67 positive cells increased from NEp through AAH and HGPIN to PCa. In particular, the mean Hscore of PTOV1 expression in AAH was 110.90, i.e., close to three times that of NEp (40.76), similar to that of HGPIN (105.61) and lower than that of PCa (137.03). The mean values in AAH and HGPIN associated with cancer in the RPs were slightly higher than in the SPs. Our findings related to PTOV1 expression in AAH, similar to those in HGPIN, provide additional evidence linking AAH to prostatic adenocarcinoma.

A new method to enhance peripheral nerve regeneration is described. A 2-cm-long segment of right sciatic nerve of 90 Sprague-Dawley rats was transected and the gap was then bridged using one of the three methods: group 1 (treated, n 1 = 30), a 2-cm-long strip of perineurium inserted inside a 2-cm-long vein graft; group 2 (control, n 2 = 30), a vein graft alone; group 3 (standard nerve graft, n 3 = 30), a resected segment of the sciatic nerve. Clinical and electrophysiological tests and quantitative analysis carried out 12 weeks after surgical procedure showed a good nerve regeneration for the treated and the control vein group, although it was lower than the standard nerve graft group. G factor values, index of nervous myelinization, were not significantly different between groups 1 and 3 (P

Various findings in prostate tissue indicate raised risk of later prostate cancer, and, of all these, the finding of high-grade prostatic intraepithelial neoplasia on repeated biopsy is most strongly associated. This Review discusses the morphologic spectrum and clinical importance of these proposed preneoplastic lesions and conditions of the prostate, especially high-grade prostatic intraepithelial neoplasia. High-grade prostatic intraepithelial neoplasia (HGPIN) is the most likely precursor of prostatic adenocarcinoma according to virtually all available evidence. This lesion is characterized by cellular proliferations within pre-existing ducts and acini, with nuclear and nucleolar enlargements similar to those seen in prostate cancer, although unlike cancer HGPIN retains a basal-cell layer. The recognition of HGPIN is clinically important because of the strong association between this disease and prostatic carcinoma. The predictive value for cancer of an initial diagnosis of HGPIN on needle biopsy has substantially declined, with values falling from 36% to 21%. A major factor contributing to this decline is related to increased use of needle biopsy core sampling, which has provided the means for many cancers associated with HGPIN to be detected on initial biopsy; repeat biopsy, even with good sampling, does not detect many additional cancers. Other possible findings in the prostate might indicate premalignant disease (low-grade prostatic intraepithelial neoplasia, atrophy, malignancy-associated changes, and atypical adenomatous hyperplasia or adenosis), but the data for these premalignant diseases are much less convincing than those for HGPIN.

Subtle morphological changes and molecular alterations have been reported in normal-appearing tissue in prostates with high-grade prostatic intraepithelial neoplasia (PIN) and prostate cancer (PCa). The severity and the distribution of these changes and alterations within the prostate gland have not been addressed in previous publications. The aim of this study was to investigate morphometrically the nuclear changes of the normal-looking columnar epithelium adjacent to and distant from high-grade PIN and PCa. Karyometry was performed on the whole-mount histological sections of three radical prostatectomy (RP) specimens. Two concentrical lines, one corresponding to the outer surface (or capsule) of the prostate and the other corresponding to one centimeter towards the center, were drawn with a black pen on each whole-mount section. The part of the prostate tissue between these two boundaries was then divided into twelve equal sectors or regions. The part within the inner line was divided into two regions. The analysis was also performed on the slides of the apex and base of the prostate. One prostate contained normal-looking epithelium only (case no. 1). Another contained both high-grade PIN and PCa, the former occupying larger areas than the latter (case no. 2). Both high-grade PIN and PCa were present in the third sample, in which PCa was more widely distributed than PIN (case no. 3). The lesion measured in each region was always the most severe, e.g., either high-grade PIN or PCa. When neither were identifiable, then the normal-looking columnar epithelium was analyzed. For each sector, the mean and standard deviation of the nuclear area, maximum nuclear diameter, nuclear roundness factor, and nucleolar area were calculated. In normal-looking columnar epithelium, the mean of the mean nuclear area of the sectors of case no. 1 was 35.19 µm2 (SD 4.14). The mean nuclear areas in cases no. 2 and no. 3 were 37.94 µm2 (SD 4.65) and 37.31 µm2 (SD 4.36), respectively. The mean of the mean nuclear area of the sectors with high-grade PIN of case no. 2 was 49.85 µm2 (SD 8.44), whereas it was 54.26 µm2 (SD 2.91) in case no. 3. The mean of the nuclear area values obtained in the sectors of cases no. 2 and no. 3 with PCa was 56.74 µm2 (SD 6.56) and 61.17 µm2 (SD 8.13), respectively. When considering the normal-looking tissue of the second and third case, 79% and 90%, respectively, of the regions showed nuclear area values greater than 34.94 µm2 (e.g., the 50th percentile of the mean nuclear area values of the regions of the first case). Sectors with normal-looking epithelium, whose nuclear area was above this threshold, were both adjacent to and at a distance of more than 1 cm from those with PIN or PCa. The other nuclear features showed a similar trend of value changes. This study demonstrates that the normal-looking ducts and acini from prostate harboring preneoplastic and neoplastic lesions show morphological nuclear abnormalities that are not seen by the human eyes but that can be detected with image analysis. Such changes may be of diagnostic importance, especially in cases where clinical suspicion for cancer prevails after a negative biopsy.

Various grading systems have been proposed for renal cell carcinoma (RCC), using nuclear, cytoplasmic, and architectural features. The available evidence suggests that nuclear grading is a better prognostic indicator than other types of grading schemes. Nuclear morphometry may still improve the correlation of the nuclear grading with survival, however, because observer consistency is lacking in the subjective grading of RCC. The aim of this study was to investigate morphometrically whether RCC cases show a continuous spectrum of nuclear changes or whether there are discrete groups of cancer that correspond to the four Fuhrman grades. Karyometry was performed on 5- µm-thick, haematoxylin- and eosin-stained sections from 60 cases of conventional (clear cell) RCC. The analysis also included the evaluation of normal renal tissue (proximal tubules) adjacent to cancer. In each case the difference between the value of the cancer and the corresponding normal epithelium was calculated to represent, quantitatively, the degree of similarity between the tumour tissue and the internal normal control. When the differences were sorted into ascending order, a steady increase in values was observed for both the nuclear and the nucleolar features. A monotonic trend was evident for the differences in the mean maximum nuclear diameter and mean nucleolar area. When the differences between the values in the cancer and in the corresponding normal epithelium of these two features were summed up, the method resulted in a continuous variable, or nuclear morphometric index, related to the degree of deviation of each individual RCC from its internal normal control. The lowest index values were observed in of Fuhrman grade I cases, whereas values ranging from 2.679 to 5.422 were associated with cases graded II. Values equal to or higher than 5.951 were seen in the cases assigned to either grade III or grade IV. Partial overlap was present between the index values in grades III and IV. The RCC cases can be represented by a continuous index that corresponds to the morphological grading based on the Fuhrman scheme. This study shows that the index may be useful in supplementing the pathologist’s grading. This issue can be further addressed with follow-up studies.

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 were 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.