Atypical Mycobacterial Infection Presenting as Persistent Skin Lesion in a Patient with Ulcerative Colitis

Immunosuppressive drugs are commonly used for the treatment of inflammatory bowel disease. Patients receiving immunosuppressants are susceptible to a variety of infections with opportunistic pathogens. We present a case of skin infection with Mycobacterium chelonae in a 60-year-old Caucasian woman with ulcerative colitis who had been treated with corticosteroids and azathioprine. The disease manifested with fever and rash involving the right leg. Infliximab was administered due to a presumptive diagnosis of pyoderma gangrenosum, leading to worsening of the clinical syndrome and admission to our hospital. Routine cultures from various sites were all negative. However, Ziehl-Neelsen staining of pus from the lesions revealed acid-fast bacilli, and culture yielded a rapidly growing mycobacterium further identified as M. chelonae. The patient responded to a clarithromycin-based regimen. Clinicians should be aware of skin lesions caused by atypical mycobacteria in immunocompromised patients with inflammatory bowel disease. Furthermore, they should be able to thoroughly investigate and promptly treat these conditions

Study of the role of natural antimicrobial peptides in the pathogenesis of infections in cirrhotic patients

Objective: To investigate the expression of natural antimicrobial peptides in chronic liver disease and the presence or not of a correlation with the etiology of liver disease. Moreover, to elucidate the correlation with factors related to bacterial translocation and the possible role of antimicrobial peptides (AMPs) in the pathogenesis of infections in cirrhotic patients and/or the role of AMPs as a biomarker of bacterial translocation in patients with end stage liver disease. Patients and Methods: The concentrations of hBD-1 and sCD14 in plasma and sera of patients with liver cirrhosis, chronic viral hepatitis and healthy controls, and in hepatic veins of cirrhotic patients, were measured by ELISA. The relative mRNA expression of various natural antimicrobial peptides was determined by Real-time RT-PCR in biopsies from the terminal ileum and colon. Moreover, experiments concerning immunohistochemistry and blood and tissue cultures were performed. Results: hBD-1 and sCD14 were significantly upregulated in peripheral blood of cirrhotic patients (18.26 ng/ml, 16.65-21.94, median, 95%CI for hBD-1 and 2777 ng/ml, 2701-3133, median, 95%CI for sCD14) compared to healthy controls (8.85 ng/ml, 7.46-9.69, median, 95%CI, P<0.0001 for hBD-1 and 1671 ng/ml, 1574-2026, median, 95%CI, P<0.0001 for sCD14) and concerning hBD-1 to patients with chronic viral hepatitis too (8.87 ng/ml, 9.09-12.88, median, 95%CI, P<0.0001).. The etiology of cirrhosis did not affect the concentration of either protein. The levels of hBD-1 protein correlated significantly with the levels of sCD14 in blood collected from hepatic veins of cirrhotic patients (Spearman r=0.6, P=0.0045). In contrast, no significant differences were observed in the intestinal mucosal mRNA expression of the Paneth cell specific defensin A5 or hBD-1 between patients with cirrhosis and healthy controls. Conclusions: hBD-1 is upregulated in patients with cirrhosis and highly correlates with LPS-induced protein, sCD14. The correlation of hBD-1 with the presence of gram negative bacteremia indicates a possible role of hBD-1 as a biomarker of bacterial translocation in patients with cirrhosis.Σκοπός: Η μελέτη της έκφρασης των φυσικών αντιμικροβιακών πεπτιδίων στη χρόνια ηπατική νόσο και η ύπαρξη ή όχι συσχέτισης με την αιτιολογία της ηπατικής νόσου. Επιπλέον, η συσχέτισή τους με παράγοντες που σχετίζονται με τη βακτηριακή αλλόθεση και η ανάδειξη του πιθανού ρόλου των αντιμικροβιακών πετιδίων στη παθογένεση των λοιμώξεων στους κιρρωτικούς ασθενείς ή/και του ρόλου τους ως δείκτη βακτηριακής αλλόθεσης στους ασθενείς με τελικού σταδίου ηπατική νόσο.Ασθενείς και Μέθοδοι: Οι συγκεντρώσεις της hBD-1 και του sCD14 στο πλάσμα και στον ορό αντίστοιχα ασθενών με κίρρωση του ήπατος, με χρόνια ιογενή ηπατίτιδα και σε υγιείς μάρτυρες, καθώς και στις ηπατικές φλέβες κιρρωτικών ασθενών, μετρήθηκαν με ELISA. Η σχετική έκφραση του mRNA διαφόρων αντιμικροβιακών πεπτιδίων προσδιορίστηκε Real-time RT-PCR σε βιοψίες από τον τελικό ειλεό και το παχύ έντερο. Επίσης, διενεργήθηκαν πειράματα ανοσοϊστοχημείας και καλλιεργειών σε ιστικά τεμαχίδια και πειράματα καλλιεργειών ολικού αίματος.Αποτελέσματα: Παρατηρήθηκε σημαντική αύξηση της hBD-1 και του sCD14 στο περιφερικό αίμα των ασθενών με κίρρωση του ήπατος (18.26 ng/ml, 16.65-21.94, διάμεσος, 95%CI για την hBD-1 και 2777 ng/ml, 2701-3133, διάμεσος, 95%CI για το sCD14) σε σύγκριση με τους υγιείς μάρτυρες (8.85 ng/ml, 7.46-9.69, διάμεσος, 95%CI, P<0.0001 για την hBD-1 και 1671 ng/ml, 1574-2026, διάμεσος, 95%CI, P<0.0001 για το sCD14) και όσον αφορά στην hBD-1 και με τους ασθενείς με χρόνια ιογενή ηπατίτιδα (8.87 ng/ml, 9.09-12.88, διάμεσος, 95%CI, P<0.0001). Η υποκείμενη αιτία της κίρρωσης δεν επηρέαζε τη συγκέντρωση καμίας εκ των δύο πρωτεϊνών. Τα επίπεδα της hBD-1 παρουσιάζουν στατιστικά σημαντική συσχέτιση με τα επίπεδα του sCD14 στον ορό που συλλέχθηκε από τις ηπατικές φλέβες των κιρρωτικών ασθενών (Spearman r=0.6, P=0.0045). Αντιθέτως, δεν παρατηρήθηκαν διαφορές μεταξύ κιρρωτικών ασθενών και υγιών μαρτύρων στην τοπική έκφραση του mRNA ούτε της A5 ντεφενσίνης ή της hBD-1 στο εντερικό επιθήλιο.Συμπεράσματα: Η hBD-1 υπερεκφράζεται στους ασθενείς με κίρρωση του ήπατος και σχετίζεται σε στατιστικά σημαντικό επίπεδο με το sCD14, ενός μορίου του οποίου η έκφραση επάγεται από την παρουσία του λιποπολυσακχαρίτη (LPS). Η συσχέτιση της hBD-1 με την παρουσία μικροβιαιμίας από gram αρνητικά μικρόβια, αναδεικνύει τον πιθανό ρόλο της hBD-1 ως δείκτη βακτηριακής αλλόθεσης σε ασθενείς με κίρρωση του ήπατος

High intestinal and systemic levels of decoy receptor 3 (DcR3) and its ligand TL1A in active ulcerative colitis

Decoy receptor-3 (DcR3) is a member of the TNF receptor superfamily of proteins, which has been implicated in anti-apoptotic and anti-inflammatory pathways, via binding to TL1A, LIGHT and Fas-L. The role of the TL1A/DcR3 ligand/receptor pair in ulcerative colitis (UC) has not been studied. We investigated the systemic (peripheral blood) and local (large intestine) expression of DcR3 and TL1A in 64 patients with UC and 56 healthy controls. DcR3 serum concentrations were highly elevated in patients with active UC (P &lt; 0.0001 vs. healthy controls). This elevation was clearly related to the presence of intestinal inflammation as it was less frequently observed in patients in remission (P=0.003 vs. active UC) whereas effective treatment resulted in disappearance or significant decrease of serum DcR3 (P=0.006 vs. pre-treatment). Furthermore, DcR3 mRNA transcripts were significantly elevated in inflamed areas of the colon (P=0.002 vs. non-affected of the same patient). In addition to DcR3 elevation, we found increased circulating levels of TL1A in patients with either active or inactive UC in comparison to healthy controls (P &lt; 0.001 for both). We conclude that elevated serum DcR3 may serve as an indicator of active colonic inflammation in patients with UC. TL1A/DcR3-mediated pathways may participate in the pathogenesis of UC. (C) 2010 Elsevier Inc. All rights reserved

Differential expression of the TL1A/DcR3 system of TNF/TNFR-like proteins in large vs. small intestinal Crohn&apos;s disease

Background: TNF-like cytokine 1A provides co-stimulatory signals to activated lymphocytes through binding to death-domain receptor-3. Decoy receptor-3 inhibits death-domain receptor-3 signalling, rendering immunocytes resistant to apoptosis. These functions may be important for the pathogenesis of Crohn’s disease. Aims: To study the mucosal and systemic expression of Decoy receptor-3 and TNF-like cytokine 1A in Crohn’s disease, in relation to disease activity, localization, and response to treatment. Methods: Soluble Decoy receptor-3 and TNF-like cytokine 1A were measured by ELISA in active or quiescent Crohn’s disease. Relative mRNA expression in non-affected and inflamed intestinal mucosa was determined by real-time RT-PCR. Results: We found significant upregulation of Decoy receptor-3 and its ligands TNF-like cytokine 1A and FasL in inflamed intestinal mucosa of Crohn’s disease patients. During active disease, Decoy receptor-3 and TNF-like cytokine 1A were detected in the serum in the majority of patients. Intestinal inflammation was strongly associated with these elevations as they were absent during remission and significantly reduced with anti-inflammatory treatment. Regional diversity was observed as Decoy receptor-3 was upregulated in colonic and ileal sites, whereas TNF-like cytokine 1A was preferentially induced in the large bowel mucosa and systemic circulation of patients with colonic involvement. Conclusions: TNF-like cytokine 1A and Decoy receptor-3 are upregulated during active Crohn’s disease and may participate in disease pathogenesis and offer novel therapeutic opportunities. (C) 2011 Editrice Gastroenterologica ltaliana S.r.l. Published by Elsevier Ltd. All rights reserved

Transgelin Up-Regulation in Obstructive Nephropathy

<div><p>Fibrosis is a complex and multifactorial process, affecting the structure and compromising the function of several organs. Among those, renal fibrosis is an important pathological change, eventually leading to renal failure. Proteomic analysis of the renal parenchyma in the well-established rat model of unilateral ureteral obstruction (UUO model) suggested that transgelin was up-regulated during the development of fibrosis. Transgelin up-regulation was confirmed both at the protein and at the mRNA level. It was observed that at early stages of fibrosis transgelin was mainly expressed in the interstitial compartment and, more specifically, in cells surrounding the glomeruli. Subsequently, it was confirmed that transgelin expressing cells were activated fibroblasts, based on their extensive co-expression of α-SMA and their complete lack of co-distribution with markers of other cell types (endothelial, epithelial and cells of the immune system). These periglomerular fibroblasts exhibited staining for transgelin mainly cytoplasmic but occasionally nuclear as well. In addition, transgelin expression in periglomerular fibroblasts was absent in renal fibrosis developed in a hypertensive model, compared to the UUO model. Promoter analysis indicated that there are several conserved motifs for transcription factor binding. Among those, Kruppel-like factor 6 was found to be up-regulated in transgelin positive periglomerular activated fibroblasts, suggesting a possible involvement in the mechanism of transgelin up-regulation. These data strongly suggest that transgelin is up-regulated in the obstructive nephropathy and could be used as a novel marker for renal fibrosis in the future.</p></div

Nuclear localization of transgelin in the UUO model.

<p>Immunohistochemistry for transgelin in the ligated animals sacrificed after 2 days (A) and 8 days (B) of ligation. Immunofluorescence for transgelin in the ligated animals sacrificed after 2 days (C) and 8 days (D) after ligation. In 6E, a selected glomerulus under a 63x/1.4NA lens of confocal microscopy is seen, where transgelin is represented with green and the DAPI stains the nuclei (blue). In 6F (which splits in F1- transgelin/green chromophore and F2- DAPI/blue chromophore), a magnified area from the glomerulus of 6E is represented.</p

The expression of transgelin in the UUO model at the protein level.

<p>A) Western Blot Analysis of transgelin in sham-operated and ligated animals sacrificed 2 and 8 days after ligation in rats. B) Quantification of Western Blot Analysis in rat UUO model. C) Western Blot Analysis of transgelin in sham-operated and ligated animals sacrificed 2 and 8 days after ligation in mice. D) Quantification of Western Blot Analysis in mice UUO model. In B and D, the asterisk denotes a p value <0.05.</p

Proteomic analysis in rat UUO model.

<p>2D gel electrophoresis from proteomic analysis of sham-operated (A) and ligated animals (B) sacrificed 8 days after ligation.</p