Contribution of Extracellular Matrix and Signal Mechanotransduction to Epithelial Cell Damage in Inflammatory Bowel Disease Patients: A Proteomic Study

This study utilizes 2D-DIGE (difference gel etrophoresis), isotope-coded protein labeling and biochemical assays to characterize protein alteration in ulcerative colitis (UC) and Crohn's disease (CD) in human epithelial cell and mucosal biopsies in inflammatory bowel disease (IBD)-affected patients. The aim of this study is to identify the key molecular signatures involved in epithelial cell structure of IBDs. In non-inflamed UC (QUC) keratins, vimentin, and focal adhesion kinase (7) increased, whereas vinculin and de-tyrosinated \uce\ub1-tubulin decreased; inflammation (IUC) exacerbated molecular changes, being collagen type VI alpha 1 chain (COL6A1), tenascin-C and vimentin increased. In non-inflamed CD (QCD), tenascin C, de-tyrosinated \uce\ub1-tubulin, vinculin, FAK, and Rho-associated protein kinase 1 (ROCK1) decreased while vimentin increased. In inflamed CD (ICD), COL6A1, vimentin and integrin alpha 4 increased. In QUC, cell metabolism is characterized by a decrease of the tricarboxylic acid cycle enzymes and a decrease of short/branched chain specific acyl-CoA dehydrogenase, fatty acid synthase, proliferator-activated receptors alpha, and proliferator-activated receptors gamma. In QCD a metabolic rewiring occurs, as suggested by glycerol-3-phosphate dehydrogenase (GPD2), pyruvate dehydrogenase E1 component subunit beta, NADH dehydrogenase [ubiquinone] iron-sulfur protein 3, and 4-trimethylaminobutyraldehyde dehydrogenase increment, while dihydrolipoyl dehydrogenase decreased. Macroautophagy is activated in QUC and IUC, with increased levels of p62, HSC70, major vault protein, myosin heavy chain 9, whereas it is blunted in QCD and ICD. The differing pattern of extracellular matrix, cytoskeletal derangements, cellular metabolism, and autophagy in UC and CD may contribute to the pathophysiological understanding of these disorders and serve as diagnostic markers in IBD patients

Long-term monitoring of rec-GH treatment by serial determination of serum aminoterminal propeptide of type III procollagen in children and adults with GH deficiency

Serum aminoterminal propeptide of type III procollagen (PIIINP) levels, a reliable marker of collagen formation, were evaluated in children (C=7) and adults with childhood-onset (CO=10) and acquired (A=18) GH deficiency (GHD) before, during and after withdrawal of rec-GH therapy (C=0.6 IU/kg/week, CO=0.5 IU/kg/week, A=0.25 IU/kg/week). The duration of treatment was 12 months for C and A and 6 months for CO; investigations were carried out before and at 3, 6, 9 and 12 months (for C and A) and at 3 and 6 months (for CO) of GH treatment and 6 months after the withdrawal of therapy (for A and CO). Data obtained from patients were compared with those recorded in two age- and sex-matched control groups. Before treatment, serum PIIINP levels were significantly lower (p<0.001) in C with GHD (mean+/-SE: 2.9+/-0.4 ng/ml) than in controls (6.1+/-0.4 ng/ml), while no significant differences were recorded between adults with CO/A-GHD (3.7+/-0.5 ng/ml and 3.4+/-0.2 ng/ml) and controls (3.2+/-0.2 ng/ml). GH treatment caused a significant increase (p<0.0001) of PIIINP levels both in C (3(rd) month: 4.4+/-0.2 ng/ml, 6(th) month: 5.1+/-0.4 ng/ml, 12(th) month: 5.1+/-0.5 ng/ml), CO-GHD (3(rd) month: 12.7+/-1.2 ng/ml; 6(th) month: 10.2+/-0.6 ng/ml) and A-GHD (3(rd) month: 10.0+/-1.0 ng/ml; 6(th) month: 8.4+/-0.6 ng/ml; 12(th) month: 7.0+/-0.7 ng/ml), the increase being dose-dependent (more marked and sustained in adults with CO-GHD). The maximal stimulation of collagen synthesis occurred after 3 months of GH treatment in adults with GHD, while a more gradual and less relevant increase was observed in C with GHD. Six months after the withdrawal of GH therapy, serum PIIINP levels of adults with CO-GHD (3.6+/-0.3 ng/ml) were similar to those recorded before treatment, while in adults with A-GHD serum PIIINP levels (2.6+/-0.2 ng/ml) were significantly tower (p<0.01) than in basal condition. In conclusion, our study shows that: a) GHD is associated with a reduction of soft tissue formation in children, while it seems to exert no relevant effects in adults with GHD; b) GH therapy causes a rapid stimulation of collagen turnover, which shows a different pattern in children and adults; c) the OH-induced stimulation of collagen synthesis is rapidly removed after the withdrawal of GH treatment. For these reasons, the determination of peripheral markers of GH effects appears useful for the monitoring of Chi therapy and can contribute to assess the "tailored" substitutive dose for the individual patient

Genotipo 3020insc del gene Nod2/Cardi5 in una casistica di pazienti con malattia di Crohn trattati chirurgicamente

Specifici genotipi del gene NOD2/CARD 15. e in particolare la mutazione 3020 in sC. sono associati a malattia di Crohn (Crohn), familiare e non. Per il genotipo 3020insC sono riportate variabili prevalenze (da 1.8% a 17 .2 \ub0/t:\u2022) e correlazioni con i comportamenti della malattia , sia fistolizza nt e che stenosante . Le diverse casistiche differiscono per composizione etnica , per prevalenza della malattia familiare. per modalit\ue0 di reclutamento e trattamento dei pazie nti. Scopi: in pazienti con Crohn sottoposti a trattamento chirurgico , determinare la prevalenza del genotipo 3020insC e quindi valutare se questo correli con specifiche caratteristiche clinico-patologiche.Pseudomixoma peritonei (PMP) is a rare neoplasia with a low grade of clinical malignity. Generally, the main treatment of this tumor is the surgical debulking. Best results are obtained combining surgery and hyperthermic antiblastic peritoneal perfusion (HAPP) with CDDP and MMC. From April '97 to March 2003, we operated on 132 patients, 8 times with a palliative intent. In 27 times we achieved a complete cytoreduction (17 CC0 and 10 CC1) followed by HAPP. As regards results, no post-operative mortality was reported and 19% of major morbidity was observed. 26 patients are NED at maximum follow-up of 6 years and 1 patient had recurrence 6 months after primary resection. We believe that cytoreduction and HAPP is the golden standard of PMP therapy when it is possible to achieve a complete cytoreduction. Most of the times, the disease is not radically treated and therefore, after diagnosis, patients should be only referred to specialized centers

Onset of inflammatory bowel disease during combined alpha-interferon and ribavirin therapy for chronic hepatitis C: report of two cases

We report two patients who developed an inflammatory bowel disease (IBD) shortly after beginning combined alpha-interferon and ribavirin treatment for HCV-related chronic hepatitis. The previous clinical history was negative for IBD in both patients, who developed diarrhoea and rectal bleeding 10 days and 6 months, respectively, after the initiation of therapy. The history, therapeutic management and the possible causal relationships of these cases are discussed

Histopathological and molecular studies in patients with goiter and hypercalcitoninemia : reactive or neoplastic C-cell hyperplasia?

The cut-off values able to differentiate between reactive or neoplastic C-cell hyperplasia (CCH) or to predict sporadic medullary thyroid cancer (MTC) are still debated both for basal and stimulated calcitonin (bCT and sCT). In the present study, the prevalence and the histological patterns of CCH in 15 patients with multinodular goiter (MNG), bCT>10 pg/ml and sCT levels >50 pg/ml were studied. As controls, 16 patients with MNG and bCT levels 50 pg/ml were associated with CCH or MTC, being the total number of C-cells/60 fields significantly higher than that found in MNG with normal bCT (P=0.0008) and comparable with that detected in FMTCs. In the group with sCT>50 pg/ml, the C-cells displayed a neoplastic phenotype. Neither germline nor somatic RET mutations were found. In conclusion, sCT levels >50 pg/ml were always associated with CCH, without correlation between CT levels and the number of C-cells or the final diagnosis. The C-cells had a morphology and distribution pattern similar to those observed in FMTC. Thus, sCT levels >50 pg/ml indicate the presence of CCH with a possible preneoplastic potential, suggesting the opportunity to perform a prophylactic surgical treatment

Nephron endowment assessment in pre-transplantation kidneys using a digital histomorphometric approach

Introduction Kidney function largely depends on nephron number. However, the role of (donor\u2019s) nephron endowment (NE) has been poorly investigated in kidney transplantation (KTx) probably because technically difficult and no reliable surrogate markers exists. The aim of this study was developing an easy and reliable algorithm for computation of NE in kidneys addressed to KTx. Materials and methods Fourty-one kidneys removed from 26 brain-dead donors (12 female, mean age of 62.6 years) were analyzed. Computed tomography was performed on kidneys to stereologically compute the cortical volume. Tissue sections from 6 standardized areas of each kidney were harvested and processed for histomorphometry. On each section stereological method was employed to evaluate glomerular density (GD), size and volume. NE was computed for each kidney by multiplying the cortical volume for GD and dividing by the average glomerular volume. Number of functional (FG) and atrophic (AG) glomeruli was also assessed. Results Mean volume of the cortical portion was 48.2cm3\ub117.7. Glomeruli represented 9.53%\ub12.19 of cortex. Mean glomerular volume was 5.75E-6\ub12.04E-06 cm3. Glomerular number was: 858,550\ub1373,245 (NE), 650,606\ub1310,400 (FG) and 207,944\ub199,837 (AG). GD was higher at the upper pole compared to medium and inferior areas but no significantly. Conclusions Nephron count is feasible with relatively limited resources by measuring GD in a single spot and applying the algorithm we have developed. The integration of traditional pathology with cutting-edge digital technologies and bioinformatics is a reproducible promising tool for diagnostics also in the area of KTx towards a better understanding of the mechanisms influencing the outcom

Late lung retransplantation using extracorporeal membrane oxygenation as a bridge: case report

Lung retransplantation is the only therapeutic option for acute and chronic graft failure, but only a few cases have been described to have been performed with extracorporeal membrane oxygenation (ECMO) support. We describe the successful case of a 46-year-old man treated with right lung transplantation and left lung retransplantation supported by venovenous ECMO. Lung retransplantation is the only therapeutic option to treat severe primary graft dysfunction, major technical problems, and refractory chronic rejection following pulmonary transplantation. Despite a number of comprehensive studies on lung retransplantation, only a few works have addressed the use of extracorporeal membrane oxygenation (ECMO) as a bridge to the surgical reoperation.(1-4) Herein we have presented a patient treated with pulmonary bilateral retransplantation subsequent to ECMO therapy for progressive deterioration of pulmonary function in single lung transplantation