Silvia Basato

Plasma lipids and inflammation in active inflammatory bowel diseases

Background  Ulcerative colitis (UC) and Crohn’s disease (CD) can cause metabolic and inflammatory alterations.

Aberrant gene methylation in non-neoplastic mucosa as a predictive marker of ulcerative colitis-associated CRC.

Background Promoter hypermethylation plays a major role in cancer through transcriptional silencing of critical genes. The aim of our study is to evaluate the methylation status of these genes in the colonic mucosa without dysplasia or adenocarcinoma at the different steps of sporadic and UC-related carcinogenesis and to investigate the possible role of genomic methylation as a marker of CRC. Results The expression of Dnmts 1 and 3A was significantly increased in UC-related carcinogenesis compared to non inflammatory colorectal carcinogenesis. In non-neoplastic colonic mucosa, the number of methylated genes resulted significantly higher in patients with CRC and in those with UC-related CRC compared to the HC and UC patients and patients with dysplastic lesion of the colon. The number of methylated genes in non-neoplastic colonic mucosa predicted the presence of CRC with good accuracy either in non inflammatory and inflammatory related CRC. Methods Colonic mucosal samples were collected from healthy subjects (HC) (n = 30) and from patients with ulcerative colitis (UC) (n = 29), UC and dysplasia (n = 14), UC and cancer (n = 10), dysplastic adenoma (n = 14), and colon adenocarcinoma (n = 10). DNA methyltransferases-1, -3a, -3b, mRNA expression were quantified by real time qRT-PCR. The methylation status of CDH13, APC, MLH1, MGMT1 and RUNX3 gene promoters was assessed by methylation-specific PCR. Conclusions Methylation status of APC, CDH13, MGMT, MLH1 and RUNX3 in the non-neoplastic mucosa may be used as a marker of CRC: these preliminary results could allow for the adjustment of a patients surveillance interval and to select UC patients who should undergo intensive surveillance.

Mismatch repair gene defects in sporadic colorectal cancer enhance immune surveillance

Background There is evidence that colorectal cancers (CRC) with DNA mismatch repair deficiency (MMR-D) are associated with a better prognosis than the generality of large bowel malignancies. Since an active immune surveillance process has been demonstrated to influence CRC outcome, we investigated whether MMR-D can enhance the immune response in CRC. Patients and Methods A group of 113 consecutive patients operated for CRC (42 stage I or II and 71 with stage III or IV) was retrospectively analyzed. The expression of MMR genes (MSH2, MLH1, MSH6 and PSM2) and co-stimulatory molecule CD80 was assessed by tissue microarray immunohistochemistry. In addition, tumor infiltrating mononuclear cells (TIMC) and T cell subpopulations (CD4, CD8, T-bet and FoxP-3) were quantified. The effect of specific siRNA (siMSH2, siMLH1, siMSH6 and siPSM2) transfection in HT29 on CD80 expression was quantified by flow cytometry. Non parametric statistics and survival analysis were used. Results Patients with MMR-D showed a higher T-bet/CD4 ratio (p = 0.02), a higher rate of CD80 expression and CD8 lymphocyte infiltration compared to those with no MMR-D. Moreover, in the MMR-D group, the Treg marker FoxP-3 was not expressed (p = 0.05). MMR-D patients with stage I or II and T-bet expression had a significant better survival (p = 0.009). Silencing of MSH2, MLH1 and MSH6, but not PSM2, significantly increased the rate of CD80+ HT29 cells (p = 0.007, p = 0.023 and p = 0.015, respectively). Conclusions CRC with MMR-D showed a higher CD80 expression, and CD8+ and Th1 T-cell infiltration. In vitro silencing of MSH2, MLH1 and MSH6 significantly increased CD80+ cell rate. These results suggest an enhanced immune surveillance mechanism in presence of MMR-D.

Restorative Proctocolectomy for Ulcerative Colitis: Impact on Lipid Metabolism and Adipose Tissue and Serum Fatty Acids

The aim of this prospective study was to evaluate the changes of the metabolism of circulating and storage lipids in patients with ulcerative colitis after restorative proctocolectomy. Fifteen consecutive patients and 15 sex- and age-matched healthy controls were enrolled. Disease activity, diet, inflammatory parameters, plasma lipoprotein concentrations, and fatty acids (FA) of serum phospholipids and of the subcutaneous adipose tissue were assessed at colectomy and at ileostomy closure. In ulcerative colitis patients, total cholesterol and docosahexaenoic acid were lower than in healthy subjects (p < 0.01 and p < 0.05). The median interval between colectomy and ileostomy closure was 6 (range 2–9) months. During that interval, the inflammatory parameters improved, high-density lipoproteins (HDL) cholesterol increased (p < 0.01), and low-density (LDL) cholesterol decreased (p = 0.01). At ileostomy closure, serum arachidonic acid levels were increased (p = 0.04), whereas serum oleic acid level was decreased (p = 0.02). In this interval, no significant alteration, either in serum n-3 FA precursors or in the FA of subcutaneous adipose tissue, was observed. The increase of serum arachidonic acid after colectomy might suggest a lower utilization for inflammatory process. The reduction of LDL cholesterol is an index of malabsorption probably due to the accelerated transit and to the exclusion of the terminal ileum caused by the covering ileostomy.

CD80 down-regulation is associated to aberrant DNA methylation in non-inflammatory colon carcinogenesis

BackgroundThe lack of positive costimulatory molecules represents one of the mechanisms by which tumor cells evade immune surveillance. Promoter hypermethylation plays a major role in cancer development through transcriptional silencing of critical genes. The aim of this study was to examine the expression of the costimulatory molecule CD80 in relationship with genomic methylation in non-inflammatory colon carcinogenesis.MethodsColonic mucosal samples were collected from healthy subjects (n = 30) and from dysplastic adenoma (n = 14), and colon adenocarcinoma (n = 10). DNA methyltransferases-1, −3a, −3b and CD80 mRNA expression were quantified by real time qRT-PCR. The methylation status of CDH13, APC, MLH1, MGMT1 and RUNX3 gene promoters was assessed by methylation-specific PCR. CD80 expression was assessed in HT29, HCT-15 and LoVo cell lines after treatment with the DNA-methyltransferase inhibitor 5-Aza-2′-deoxycytidine.ResultsCD80 mRNA levels were significantly lower in the non-inflammatory dysplastic colonic mucosa of patients with one or more methylated genes and inversely correlated with patients’ methylation scores (τ = −0.41, p = 0.05 and τ = −0.37, p = 0.05, respectively). Treatment with 5-Aza-2′-deoxycytidine significantly increased CD80 expression both in terms of the level of CD80 mRNA (p = 0.007) and of CD80+ cells (p = 0.003).ConclusionsThese results indicate that the failure of immune surveillance mechanisms in non-inflammatory colon carcinogenesis may be linked to genomic methylation directly or indirectly affecting CD80 expression.

Lipid and phospholipid profile after bowel resection for Crohn’s disease

Background and aimChronic inflammation, impaired intestinal adsorption, and bowel resection may have an impact on lipid metabolism before and after intestinal surgery for Crohn’s disease (CD). The aim of this prospective study was to define the impact of intestinal surgery for CD on plasma phospholipid fatty acid (FA) composition and of serum plasma lipoprotein concentrations and to investigate the role of CD recurrence on lipid parameters.Materials and methodsTwenty-four consecutive patients who had intestinal surgery for CD since December 2004 to March 2006 were enrolled in this prospective study. The total amount of calorie intake and the quality of the aliments, systemic inflammatory activity, and plasma lipoproteins and phospholipid fatty acid composition were determined at operation and at follow-up. Statistical analysis was performed with pair-matched tests.ResultsThe median follow-up was 6 (4–20) months. During the follow-up, no significant modification of body mass index was observed. An increase of high-density lipoprotein (HDL) cholesterol (p = 0.02) without other modifications in the plasma phospholipid FA composition were evidenced after surgery. The comparison between colectomy and ileo-colonic or ileal resection groups did not show any significant difference in the lipoprotein concentration and phospholipid FA profile. The length of resected bowel did not show any significant correlation with any relevant difference in lipid, phospholipid profile, or in inflammatory parameters. Patients who experienced a recurrence of CD reported significantly higher levels of total (p < 0.01), HDL (p = 0.01), and low-density lipoprotein cholesterol (p = 0.01) were observed in patients in remission than in those with recurrent active disease.ConclusionsPatients who are submitted to intestinal resection for CD improve their inflammatory status as well as their lipid metabolism, and CD recurrence, but not the extent of bowel resection, is the main predictor of alteration of serum lipid concentration.

Su1923 Immune Microenvironment in Colonic Carciongenesis: Sporadic Mismatch Repair Genes Defects Are Associated to Hicd80+ Lamina Propria Monuclear Cells Infiltration

BACKGROUND Genomic defects in DNA mismatch repair (MMR) genes (MSH2, MLH1, PSM2 or MSH6) characterize the hereditary non polyposis colon cancer (HNPCC). However most colorectal cancers (CRC) with high-frequency microsatellite instability are sporadic, wherein the MMR defect develops because of inactivation of the MLH1 gene by DNA methylation. Multiple retrospective studies, including a population-based study and a metaanalysis, have demonstrated that patients with MMR-deficient colon cancers have a more favourable stage-adjusted prognosis compared with patients whose tumors have intact MMR. Since the immune environment of CRC has been demonstrated to influence its prognosis, we aimed to investigate the interplay between MMR genes and the immune environment in CRC PATIENTS AND METHODS A group of 98 consecutive patients operated on for colorectal cancer was retrospectively analysed. Familial and medical history were retrieved to assess the presence of Bethesda criteria for HNPCC diagnosis. Immunohistochemistry for CD80, TLR4, MyD88, MSH2, MLH1, MSH6 and PSM2 was performed on tissue sections. Moreover, lamina propria mononuclear cells (LPMC), polymorphonuclear cells and eosoinophil tumour infiltration was quantified. Patients were stratified in three groups: no MMR genes defect, MMR gene defects alone and MMR genes defects and at least one positive Bethesda criteria. Non parametric statistics was used. RESULTS In the three groups no difference was observed in term of polymorphonuclear cells and eosinophil infiltration and in term of TLR4 and MyD88 expression. On the contrary, LPMC infiltration was significantly higher in patients who had a MMR gene defect alone compared to patients who had no MMR genes defect and to those with MMR gene defect and positive Bethesda criteria (p= 0.014). Similarly, a significantly higher frequency of patients with high CD80 expression was observed in patients who had a MMR genes defect alone compared to patients who had no MMR gene defect and to those with MMR genes defect and positive Bethesda criteria (p=0.048). CONCLUSION In patients with MMR defects and no Bethesda criteria for HNPCC antigen presenting cells function seems enhanced as shown by higher frequency of hiCD80+ patients and higher LPMC infiltration. This immune activation may play a role in the prognosis of these patients.

The role of early surgery in the treatment of acute diabetic foot in the elderly: a retrospective review

XXI Annual Meeting of The Italian Society of Geriatric Surgery Francesco Sciannameo, Giammario Giustozzi and Beatrice Sensi Publication of this supplement was made possible with support from the Fondazione Cassa di Risparmio di Terni e Narni Meeting abstracts – A single PDF containing all abstracts in this Supplement is available here . http://www.biomedcentral.com/content/pdf/1471-2318-9-S1info.pdf

Difference of site in ulcer lesions of diabetic foot in the elderly

Results No differences were found in the lesions in patients younger than 65 (not statistically comparable number), while in the others (patients over 65 years old) there was a clear difference of site: in naturopathic patients the most part were found in the plantar surface of the foot (51.4%), especially in metatarsal heads area (49.1%). The ischemic group had the most frequent site in extremities of toes (66.5%), while the neuroischemic lesions were located on both plantar surfaces (51.8%). Conclusion The distribution of ulcers was statistically significant in all different elderly groups and seems to be correlated with their etiology (p < 0.0001). from XXI Annual Meeting of The Italian Society of Geriatric Surgery Terni, Italy. 4–6 December 2008

P093 Restorative Proctocolectomy for Ulcerative Colitis: Impact on Lipid Metabolism and on Tissutal and Serum Fatty Acids

The aim of this prospective study was to evaluate the changes of the metabolism of circulating and storage lipids in patients with ulcerative colitis after restorative proctocolectomy. Fifteen consecutive patients and 15 sexand age-matched healthy controls were enrolled. Disease activity, diet, inflammatory parameters, plasma lipoprotein concentrations, and fatty acids (FA) of serum phospholipids and of the subcutaneous adipose tissue were assessed at colectomy and at ileostomy closure. In ulcerative colitis patients, total cholesterol and docosahexaenoic acid were lower than in healthy subjects (p<0.01 and p<0.05). The median interval between colectomy and ileostomy closure was 6 (range 2–9) months. During that interval, the inflammatory parameters improved, high-density lipoproteins (HDL) cholesterol increased (p<0.01), and low-density (LDL) cholesterol decreased (p=0.01). At ileostomy closure, serum arachidonic acid levels were increased (p=0.04), whereas serum oleic acid level was decreased (p=0.02). In this interval, no significant alteration, either in serum n-3 FA precursors or in the FA of subcutaneous adipose tissue, was observed. The increase of serum arachidonic acid after colectomy might suggest a lower utilization for inflammatory process. The reduction of LDL cholesterol is an index of malabsorption probably due to the accelerated transit and to the exclusion of the terminal ileum caused by the covering ileostomy.