Nestin, an intermediate filament protein, has traditionally been noted for its importance as a neural stem cell marker. However, in recent years, expression of nestin has shown to be associated with general proliferation of progenitor cell populations within neoplasms. There is no reported study addressing nestin expression in T4 breast cancer patients. Thus, the aim of the present study was to investigate, through immunohistochemistry, the expression and distribution of nestin in T4 breast cancer, in order to determine its association with clinical and pathological parameters as well as with patients' outcome. Nestin was detectable in tumoral cells and in endothelial cells of blood microvessels, and it is significantly expressed in triple-negative and in inflammatory breast cancer (IBC) subgroups of T4 breast tumours. The Kaplan-Meier analysis showed that the presence of nestin in tumoral cells significantly predicted poor prognosis at 5-years survival (P=0.02) and with borderline significance at 10-years of survival (P=0.05) in T4 breast cancer patients. On the basis of these observations, we speculate that nestin expression may characterize tumours with an aggressive clinical behavior, suggesting that the presence of nestin in tumoral cells and vessels may be considered an important factor that leads to a poor prognosis. Further studies are awaited to define the biological role of nestin in the etiology of these subgroups of breast cancers.

Background Several observations suggest that genes other than HLA-B*51 might be implicated in BD susceptibility and pathogenesis. In particular, as a consequence of the strong linkage disequilibrium characterizing the MHC region, it cannot be ruled out that HLA-B*51 is included in haplotypes with other functional genes partly responsible of genetic susceptibility to BD. The distribution of HLA alleles in Sardinian natives is characterized by highly preserved haplotypes. Objectives To define the contribution of HLA genes and extended HLA haplotypes conferring susceptibility to Behçet’s Disease (BD), through analysis of Sardinian subjects. Methods Forty-five unrelated Sardinian patients with BD, diagnosed according to the ISG criteria, 45 HLA-B*51 positive and 185 unselected healthy controls were enrolled in the study. DNA samples were typed for HLA class I and class II alleles and genotyped for microsatellites (MICA-TM) and single-nucleotide polymorphisms (rs1264457 HLA–E; rs2281820 motilin; rs1799724 at -857, rs361525 at -238 TNF-alpha) spanning the HLA region. Statistical analysis was carried out using Chi-square test with Yates’ correction or Fischer’s exact test. The strength of association was estimated by calculating the Odds Ratio (OR) with 95% Confidence Interval (95% CI). A value of p<0.05 was considered statistically significant where Bonferroni’s correction was applied. The LD between pairs of loci was calculated performing a likelihood-ratio test, whose empirical distribution is obtained by a permutation procedure using the statistical software Arlequin (version 3.11, Bern, Switzerland). The significance (p<0.01) of the observed likelihood ratio is found by computing the null distribution of this ratio under the hypothesis of LD, using a permutation procedure. All markers were in Hardy-Weinberg equilibrium. Maximum-likelihood haplotype frequencies and their distribution were estimated using an Expectation-Maximization algorithm and polymorphic alleles within each investigated HLA locus were constructed into extended haplotypes. Results The HLA-B*51 allele, in particular the B*51:01, was the only allele conferring susceptibility to BD (pc=0.0042; OR =4.4; 95% CI =2.0 to 9.6), among those investigated. The B*51 allele was more frequently carried by a haplotype (A2; Cw2; B*51:01; DRB1*11; DQA1*05; DQB1*03) that reached its highest frequency in patients with BD.Linkage disequilibrium analysis showed the existence of a B*51 haplotype (A2; Cw2; B*51:01; DRB1*04; DQA1*03; DQB1*03) not associated with susceptibility to the disease (figure 1). None of the investigated microsatellites and SNPs showed a different distribution in B*51 haplotypes characterizing healthy controls and BD patients. Conclusions We confirmed that B*51 is the major genetic susceptibility factor to BD and we described a significant low frequency of A2; Cw2; B*51:01; DRB1*04; DQA1*03; DQB1*03 haplotype in our patients. It might be speculated this haplotype lacks a hypothetical co-susceptibility genetic factor to BD in Sardinian natives. ...

An early event in melanocytic tumor growth is the upregulation of Notch signaling. When an active form of Notch1 is overexpressed in primary human melanocytes, it increases cell growth, survival and invasive properties, promoting melanoma progression. Recent evidence suggested that tumor initiation and growth are driven by a subset of tumor-initiating cells termed cancer stem cells. Notch1 plays a predominant role in the maintenance of melanoblasts, including melanocyte stem cells, by preventing initiation of apoptosis. Moreover, the importance of Notch1 in the regulation of tumor angiogenesis is supported by growing evidence in various cancers. Nestin has been widely used as a marker for melanocyte stem cells as well as an angiogenic marker to evaluate neovascularity of endothelial cells in tumors. To gain an insight into the impact of Notch1 activation on the maintenance of melanocyte stem cells and angiogenesis in melanoma, the expression levels of activated Notch1 and nestin were analyzed by immunohistochemistry in 114 primary cutaneous melanomas and 35 lymph node metastases. Activated Notch1 and nestin expression was also evaluated in four dysplastic melanocytic nevi. This study provides evidence that activated Notch1 is overexpressed in cutaneous melanoma, in tumor cells as well as in microvessel endothelium, and that it can promote tumor angiogenesis. Indeed, the overexpression of activated Notch1 in both tumor and vascular endothelial cells was significantly associated with microvascular density in melanoma samples. Thus, activated Notch1 inhibitors may provide a therapeutic strategy in the treatment of melanoma by blocking tumor-associated vascularization.

These results suggest the presence in melanoma of growth mechanisms involving nestin, vimentin, and GR, similarly to that occurring in embryonic and undifferentiated cells, and may help in understanding tumour biology to provide a molecular basis for clinical therapies.

ObjectiveThe purpose of the study was to examine whether the insertion (I) and/or deletion (D) polymorphism of ACE confers susceptibility to primary pterygium in Sardinian patients in a case–control study.Methods and resultsPolymorphism genotyping was performed by nested PCR using genomic DNA extracted from the whole peripheral blood of participants with (n=251) and without (n=260) pterygium. DD, ID and II genotype frequencies were: 48%, 39% and 13%, respectively, for patients with pterygium, and 15%, 40% and 44%, respectively, for the control group. A statistically significant difference was found between the pterygium and control groups for the ACE I/D polymorphism (p<0.001). Moreover, a statistically significant difference was found between the DD and II groups (p<0.01; OR=10.49; 95% CI 6.18 to 17.79), DD+ID versus II group (p<0.01; OR=5.23; 95% CI 3.37 to 8.13) and DD versus ID groups (p<0.01; OR=3.21; 95% CI 2.04 to 5.04).ConclusionsStatistical analysis showed that the DD genotype is associated with an increased risk of developing pterygium, and with a good chance that the D allele may play an important role in the development of disease.