I distretti del Mezzogiorno

CAP. 1. Definizione e caratteristiche del distretto; CAP. 2. Come sono nati i distretti nel Mezzogiorno: fattori di successo ed insuccesso;2.1.Lo sviluppo dei distretti del Mezzogiorno: Brevi cenni storici; CAP. 3. I Distretti del Mezzogiorno; Il "Triangolo del Salotto"; Il polo calzaturiero aversano; La corsetteria di Lavello ;Il TAC del tacco Il distretto barlettano delle calzature; L'abbigliamento della Puglia centrale; La percezione del fenomeno "distretto" dei piccoli e medi imprenditori in Campania, il caso di S. Giuseppe vesuviano e Solofra; Il distretto industriale di Solofra; Conclusioni

High resolution melting analysis for a rapid identification of heterozygous and homozygous sequence changes in the MUTYH gene

Background: MUTYH-associated polyposis (MAP) is an autosomal recessive form of intestinal polyposis predisposing to colorectal carcinoma. High resolution melting analysis (HRMA) is a mutation scanning method that allows detection of heterozygous sequence changes with high sensitivity, whereas homozygosity for a nucleotide change may not lead to significant curve shape or melting temperature changes compared to homozygous wildtype samples. Therefore, HRMA has been mainly applied to the detection of mutations associated with autosomal dominant or X-linked disorders, while applications to autosomal recessive conditions are less common. Methods: MUTYH coding sequence and UTRs were analyzed by both HRMA and sequencing on 88 leukocyte genomic DNA samples. Twenty-six samples were also examined by SSCP. Experiments were performed both with and without mixing the test samples with wild-type DNA. Results: The results show that all MUTYH sequence variations, including G > C and A > T homozygous changes, can be reliably identified by HRMA when a condition of artificial heterozygosity is created by mixing test and reference DNA. HRMA had a sensitivity comparable to sequencing and higher than SSCP. Conclusions: The availability of a rapid and inexpensive method for the identification of MUTYH sequence variants is relevant for the diagnosis of colorectal cancer susceptibility, since the MAP phenotype is highly variable

The Insulin Receptor Substrate 1 (Irs1) in Intestinal Epithelial Differentiation and in Colorectal Cancer

Colorectal cancer (CRC) is associated with lifestyle factors that affect insulin/IGF signaling, of which the insulin receptor substrate 1 (IRS1) is a key transducer. We investigated expression, localization and pathologic correlations of IRS1 in cancer-uninvolved colonic epithelium, primary CRCs with paired liver metastases and in vitro polarizing Caco2 and HT29 cells. IRS1 mRNA and protein resulted higher, relative to paired mucosa, in adenomas of familial adenomatous polyposis patients and in CRCs that overexpressed c-MYC, ß-catenin, InsRß, and IGF1R. Analysis of IRS1 immunostaining in 24 cases of primary CRC with paired colonic epithelium and hepatic metastasis showed that staining intensity was significantly higher in metastases relative to both primary CRC (P<0.01) and colonic epithelium (P<0.01). Primary and metastatic CRCs, compared to colonic epithelium, contained significantly higher numbers of IRS1-positive cells (P = 0.013 and P = 0.014, respectively). Pathologic correlations in 163 primary CRCs revealed that diffuse IRS1 staining was associated with tumors combining differentiated phenotype and aggressive markers (high Ki67, p53, and ß-catenin). In Caco 2 IRS1 and InsR were maximally expressed after polarization, while IGF1R was highest in pre-polarized cells. No nuclear IRS1 was detected, while, with polarization, phosphorylated IRS1 (pIRS1) shifted from the lateral to the apical plasma membrane and was expressed in surface cells only. In HT29, that carry mutations constitutively activating survival signaling, IRS1 and IGF1R decreased with polarization, while pIRS1 localized in nuclear spots throughout the course. Overall, these data provide evidence that IRS1 is modulated according to CRC differentiation, and support a role of IRS1 in CRC progression and liver metastatization

let-7e downregulation characterizes early phase colonic adenoma in APCMin/+ mice and human FAP subjects

The crypt-villus axis represents the essential unit of the small intestine, which integrity and functions are fundamental to assure tissue and whole-body homeostasis. Disruption of pathways regulating the fine balance between proliferation and differentiation results in diseases development. Nowadays, it is well established that microRNAs (miRNAs) play a crucial role in the homeostasis maintenance and perturbation of their levels may promote tumor development. Here, by using microarray technology, we analysed the miRNAs differentially expressed between the crypt and the villus in mice ileum. The emerged miRNAs were further validated by Real Time qPCR in mouse model (ApcMin/+), human cell lines and human tissue samples (FAP) of colorectal cancer (CRC). Our results indicated that miRNAs more expressed in the villi compartment are negatively regulated in tumor specimens, thus suggesting a close association between these microRNAs and the differentiation process. Particularly, from our analysis let-7e appeared to be a promising target for possible future therapies and a valuable marker for tumor staging, being upregulated in differentiated cells and downregulated in early-stage colonic adenoma samples

CORRELATION BETWEEN THYMIDYLATE SYNTHASE (TS) mRNA EXPRESSION AND TS GENE PROMOTER POLYMORPHISMS IN PRIMARY COLORECTAL CANCER PATIENTS

Tumoral expression of TS may be a prognostic marker in colorectal cancer patients and may predict the sensitivity to 5-Fluorouracil-based chemotherapy. The TS gene promoter enhancer region contains two different polymorphisms, which can influence the TS mRNA transcriptional efficiency. The first is a polymorphism of the tandem repeat sequence (2 or 3 repeats), the second is a single nucleotide polymorphism (SNP) (G&gt;C) in the second repeat of the 3R alleles that can abolish the increased transcriptional activity of 3R alleles relative to the 2R. We studied the tumoral TS mRNA expression levels and compared this parameter with the colonic mucosa TS gene polymorphisms in 48 colorectal cancer patients undergoing surgery and receiving post-operative 5-Fluorouracil-based chemotherapy. TS mRNA levels were determined by quantitative real time reverse transcriptase-polymerase chain reaction (PCR) and TS gene polymorphism by PCR followed by electrophoresis (2R/3R polymorphism) and HaeIII restriction digestion (G&gt;C). Median TS/G6PDH expression ratio was 1.52 (range 0–10.72). TS genotype frequencies were 18.7% (2R/2R), 47.9% (2R/3R) and 33.3% (3R/3R). A trend for increased TS gene expression in patients with the 3R/3R genotype vs patients with the 2R/2R or the 2R/3R genotypes was observed (median TS/G6PDH ratio 2.2 and 1.4 for 3R/3R and 2R/2R-2R/3R groups, respectively) (p=0.07). On the basis of the SNP investigation, we stratified TS genotype into 3 sub-groups: 2R/2R, 2R/3RC, 2R/3RG sub-group 1 (n=31) vs 3RG/3RG sub-group 2 (n=6) or vs 3RG/3RC sub- group 3 (n=7). We observed a statistically significant difference in the comparison of sub-group 1 with sub-group 2 only (p=0.004). Despite the limited number of patients, our results suggest that in 3R/3R patients SNP represents the most important factor in determining TS mRNA expression levels. In conclusion the addition of SNP to the TS gene tandem repeat sequence polymorphism may provide more effective prediction of sensitivity to 5-FU-based chemotherapy

Analysis of Gene Expression Profiles Reveals Novel Correlations With the Clinical Course of Colorectal Cancer

In order to discover potential markers of prognosis in colorectal cancer (CRC) we have determined gene expression profiles, using cDNA microarrays in CRC samples obtained from 19 patients in Dukes stages C and D, with favorable clinical course (Dukes C patients, survival >5 years after surgery, group A, n=7) or unfavorable clinical course (Dukes stage C and D patients, survival <5 years after surgery, group B, n=12). Gene expression was measured in RNA from each tumor, using a pool of equal amounts of RNA from all tumors as a reference. To identify and rank differentially expressed genes we used three different analytical methods: (i) Significance Analysis of Microarrays (SAM), (ii) Cox's Proportional Hazard Model, and (iii) Trend Filter (a mathematical method for the assessment of numerical trends). The level of expression of a gene in an individual tumor was regarded as of interest when that gene was identified as differentially expressed by at least two of these three methods. By these stringent criteria we identified eight genes (ITGB2, MRPS11, NPR1, TXNL2, PHF10, PRSS8, KCNK3, JAK3) that were correlated with prolonged survival after surgery. Pathway analysis showed that patients with favorable prognosis had several activated metabolic pathways (carbon metabolism, transcription, amino acid and nitrogen metabolism, signaling and fibroblast growth factor receptor pathways). To further validate individual gene expression findings, the RNA level of each gene identified as a marker with microarrays was measured by real-time RT-PCR in CRC samples from an independent group of 55 patients. In this set of patients the Cox Proportional Hazard Model analysis demonstrated a significant association between increased patient survival and low expression of ITGB2 (p = 0.011) and NPR1 (p = 0.023) genes