Gianfranco Fanello

Exposure of Toll-Like Receptors 4 to Bacterial Lipopolysaccharide (LPS) Impairs Human Colonic Smooth Muscle Cell Function

Endotoxemia by bacterial lipopolysaccharide (LPS) has been reported to affect gut motility specifically depending on Toll‐like receptor 4 activation (TLR4). However, the direct impact of LPS ligation to TLR4 on human smooth muscle cells (HSMC) activity still remains to be elucidated. The present study shows that TLR4, its associated molecule MD2, and TLR2 are constitutively expressed on cultured HSMC and that, once activated, they impair HSMC function. The stimulation of TLR4 by LPS induced a time‐ and dose‐dependent contractile dysfunction, which was associated with a decrease of TLR2 messenger, a rearrangement of microfilament cytoskeleton and an oxidative imbalance, i.e., the formation of reactive oxygen species (ROS) together with the depletion of GSH content. An alteration of mitochondria, namely a hyperpolarization of their membrane potential, was also detected. Most of these effects were partially prevented by the NADPH oxidase inhibitor apocynin or the NFκB inhibitor MG132. Finally, a 24 h washout in LPS‐free medium almost completely restored morphofunctional and biochemical HSMC resting parameters, even if GSH levels remained significantly lower and no recovery was observed in TLR2 expression. Thus, the exposure to bacterial endotoxin directly and persistently impaired gastrointestinal smooth muscle activity indicating that HSMC actively participate to dysmotility during infective burst. The knowledge of these interactions might provide novel information on the pathogenesis of infection‐associated gut dysmotility and further clues for the development of new therapeutic strategies. J. Cell. Physiol. 223: 442–450, 2010.

Pancreaticojejunostomy with Applicationof Fibrinogen/Thrombin-Coated Collagen Patch (TachoSil®) in Roux-en-Y Reconstruction after Pancreaticoduodenectomy

To the Editor We read with great interest the article by Wellner and colleagues about the comparison between pancreaticogastrostomy (PG) and Roux-en-Y pancreaticojejunostomy (PJ) after pancreaticoduodenectomy (PD) with regard to postoperative pancreatic fistula (POPF) and other complications. The authors concluded that PG was superior to PJ in terms of clinically relevant POPF; although this study is retrospective, the use of a large case number and standardized measures in evaluation of the surgical outcome makes the results not negligible. Instead, the results of our previously described technique of Roux-en-Y reconstruction show that PJ may have a lower prevalence of POPF than that reported by Wellner and colleagues and suggest that outcome after Roux-en-Y reconstruction with regard to POPF can be further improved using fibrinogen/thrombincoated collagen patch (TachoSil®, Nycomed, UK Ltd.) in carrying out PJ. Briefly, we reviewed the clinical records of 54 consecutive patients who underwent PD by one surgeon (P.C.) at “La Sapienza” University (Rome, Italy) from January 1995 to December 2008. The underlying diseases were: pancreatic carcinoma in 31 cases; pancreatic serous cystadenoma in six cases; mucinous cystadenoma in one case; pancreatic endocrine tumor in two cases; ampullar carcinoma in seven cases; distal bile duct carcinoma in six cases; and chronic pancreatitis in one case. In all patients, the surgical procedure comprised PD with suprapyloric gastric resection and Roux -en-Y reconstruction with anastomosis of the isolated Roux limb to the stomach and single Roux limb to both the pancreatic stump and hepatic duct. Small catheters were inserted in the main duct, passed through the anastomosed bowel loop and fixed to the abdominal wall (Fig. 1a, b). A drainage tube was placed near to the pancreaticojejunostomy; external biliary drainage was not used. Pancreaticojejunal end-to-end anastomosis was done by simple invagination of the pancreatic stump into the jejunal loop for 2 cm and sutured all around with a singlelayer interrupted pledget-supported Ticron stitches between the seromuscularis of the jejunum and the pancreatic capsule. From January 2005, TachoSil® has been layered on suture line of pancreaticojejunal anastomosis (Fig. 1c, d). All 27 consecutive patients had pancreaticojejunostomy without TachoSil® (group A) whereas 27 consecutive patients had pancreaticojejunostomy with TachoSil®. All patients in our study received octreotide during the first six postoperative days. The postoperative surgical outcome within 60 postoperative days was assessed. POPF, postoperative hemorrhage J Gastrointest Surg (2009) 13:1396–1398 DOI 10.1007/s11605-009-0894-7

Reconstitution of Intestinal CD4 and Th17 T Cells in Antiretroviral Therapy Suppressed HIV-Infected Subjects: Implication for Residual Immune Activation from the Results of a Clinical Trial

Introduction During HIV infection the severe depletion of intestinal CD4+ T-cells is associated with microbial translocation, systemic immune activation, and disease progression. This study examined intestinal and peripheral CD4+ T-cell subsets reconstitution under combined antiretroviral therapy (cART), and systemic immune activation markers. Methods This longitudinal single-arm pilot study evaluates CD4+ T cells, including Th1 and Th17, in gut and blood and soluble markers for inflammation in HIV-infected individuals before (M0) and after eight (M8) months of cART. From January 2010 to December 2011, 10 HIV-1 naïve patients were screened and 9 enrolled. Blood and gut CD4+ T-cells subsets and cellular immune activation were determined by flow-cytometry and plasma soluble CD14 by ELISA. CD4+ Th17 cells were detected in gut biopsies by immunohistochemistry. Microbial translocation was measured by limulus-amebocyte-lysate assay to detect bacterial lipopolysaccharide (LPS) and PCR Real Time to detect plasma bacterial 16S rDNA. Results Eight months of cART increased intestinal CD4+ and Th17 cells and reduced levels of T-cell activation and proliferation. The magnitude of intestinal CD4+ T-cell reconstitution correlated with the reduction of plasma LPS. Importantly, the magnitude of Th17 cells reconstitution correlated directly with blood CD4+ T-cell recovery. Conclusion Short-term antiretroviral therapy resulted in a significant increase in the levels of total and Th17 CD4+ T-cells in the gut mucosa and in decline of T-cell activation. The observation that pre-treatment levels of CD4+ and of CD8+ T-cell activation are predictors of the magnitude of Th17 cell reconstitution following cART provides further rationale for an early initiation of cART in HIV-infected individuals. Trial Registration ClinicalTrials.gov NCT02097381

Probiotic supplementation promotes a reduction in T-cell activation, an increase in Th17 frequencies, and a recovery of intestinal epithelium integrity and mitochondrial morphology in ART-treated HIV-1-positive patients

HIV infection is characterized by a persistent immune activation associated to a compromised gut barrier immunity and alterations in the profile of the fecal flora linked with the progression of inflammatory symptoms. The effects of high concentration multistrain probiotic (Vivomixx®, Viale del Policlinico 155, Rome, Italy in EU; Visbiome®, Dupont, Madison, Wisconsin in USA) on several aspects of intestinal immunity in ART‐experienced HIV‐1 patients was evaluated.

Endoscopic treatment of benign and malignant strictures of the cervical esophagus and hypopharynx.

Objectives: We evaluated the efficacy of endoscopic techniques employed in the management of cervical esophageal and hypopharyngeal strictures. Methods: A series of 45 patients with cervical esophageal (35) and/or hypopharyngeal strictures (10) were included. Twenty-five patients (55.6%) with neoplastic strictures were treated for palliation alone. The stenosis was related to radiotherapy in 11 patients (24.4%) and to postsurgical complications in 9 (20%). A group of 23 patients was treated with dilation alone (group 1). A second group included 22 patients treated with insertion of a self-expandable stent after failure of dilation treatment (group 2). The swallowing test data, clinical notes, and surgical reports were reviewed. Results: All of the patients showed some degree of relief of dysphagia. In group 1, 19 of the 23 patients required multiple dilation treatments to maintain normal deglutition. In group 2, 7 of the 22 patients recovered regular oral feeding after stent placement, 10 patients reported pain and foreign body sensation, 2 patients reported pain so severe that stent removal was required, and 3 patients experienced stent migration. All but 3 of the 25 patients with inoperable tumors died during follow-up, but no patients with benign stenosis died. Conclusions: The two groups showed comparable functional results. Dilation often requires multiple procedures, but is usually well tolerated. Placement of self-expandable stents is effective, but is generally less well tolerated.

Roux-en-Y end-to-end and end-to-side double pancreaticojejunostomy: application of the reconstructive method of the Beger procedure to central pancreatectomy

PurposesCentral pancreatectomy is indicated for treatment of traumatic lesions and benign or low-grade tumors of the pancreatic neck and proximal body. After central pancreatectomy, the proximal pancreatic stump is usually closed, and pancreaticojejunostomy or pancreaticogastrostomy carried out with the distal pancreas. Adopting these reconstructive techniques in most series revealed a prevalence of postoperative fistula that was higher than after pancreaticoduodenectomy or left pancreatectomy. We present a case treated by novel application of the reconstructive method of the Beger procedure.MethodsReconstruction by Roux-en-Y double pancreaticojejunostomy after central pancreatectomy was done in a 71-year-old female suffering from insulinoma of the proximal pancreatic body.ResultsPostoperative complications were not observed. No alteration of pancreatic endocrine and exocrine function occurred at 22-month follow-up.ConclusionsDouble pancreaticojejunostomy is a promising method for treating the proximal pancreatic stump after central pancreatectomy.

Probiotics Differently Affect Gut-Associated Lymphoid Tissue Indolamine-2,3-Dioxygenase mRNA and Cerebrospinal Fluid Neopterin Levels in Antiretroviral-Treated HIV-1 Infected Patients: A Pilot Study

Recently the tryptophan pathway has been considered an important determinant of HIV-1 infected patients’ quality of life, due to the toxic effects of its metabolites on the central nervous system (CNS). Since the dysbiosis described in HIV-1 patients might be responsible for the microbial translocation, the chronic immune activation, and the altered utilization of tryptophan observed in these individuals, we speculated a correlation between high levels of immune activation markers in the cerebrospinal fluid (CSF) of HIV-1 infected patients and the over-expression of indolamine-2,3-dioxygenase (IDO) at the gut mucosal surface. In order to evaluate this issue, we measured the levels of neopterin in CSF, and the expression of IDO mRNA in gut-associated lymphoid tissue (GALT), in HIV-1-infected patients on effective combined antiretroviral therapy (cART), at baseline and after six months of probiotic dietary management. We found a significant reduction of neopterin and IDO mRNA levels after the supplementation with probiotic. Since the results for the use of adjunctive therapies to reduce the levels of immune activation markers in CSF have been disappointing so far, our pilot study showing the efficacy of this specific probiotic product should be followed by a larger confirmatory trial.

Combined radiological-endoscopic management of difficult bile duct stones: 18-year single center experience.

Objectives: Clinical evidence regarding radiological–endoscopic management of intrahepatic bile duct stones is currently lacking. Our aim is to report our 18-year experience in combined radiological–endoscopic management of intrahepatic difficult bile duct stones. Methods: From June 1994 to June 2012, 299 symptomatic patients with difficult bile duct stones were admitted to our institution. Percutaneous transhepatic cholangiography (PTC)/biliary drainage/s was performed, dilating the PTC track to 10 or 16 French within 3–7 days. Afterward we carried out percutaneous transhepatic cholangioscopy (PTCS) with electrohydraulic lithotripsy (EHL) and/or interventional radiology techniques. Follow up was made with clinical/laboratory tests and ultrasound (US). We retrospectively analyzed our radiological–endoscopic approach and reported our technical and clinical outcomes. Results: Complete stone clearance was achieved in 298 patients after a maximum of 4 consecutive sessions. Most patients (64.6%) were treated with PTCS/EHL alone, while the remaining with radiological techniques alone (26%) or a combination of both techniques (13.3%). Recurrence of stones occurred in 45 cases (15%, Tsunoda class III and class IV) within 2 years and were successfully retreated. Major adverse events were: 5 (1.6%) cases of massive bleeding that required embolisation, 2 (0.66%) perforations of the common bile duct and 31 cases (10.3%) of acute cholangitis managed with medical therapy or intervention. Conclusion: After 18 years of experience we demonstrated that our combined radiological–endoscopic approach to ‘difficult bile duct stones‘ may result in both immediate and long-term clearance of stones with a low rate of adverse events.

Inflammatory myoglandular polyp of the cecum: case report and review of literature

BackgroundInflammatory myoglandular polyp (IMGP) is a rare non-neoplastic polyp of the large bowel, commonly with a distal localization (rectosigmoid), obscure in its pathogenesis. Up till now, 60 cases of IMGP have been described in the literature, but none located in the cecum.Case presentationWe report a case of a 53-year-old man who was admitted to our hospital for further evaluation of positive fecal occult blood test associated to anemia. A colonoscopy identified a red, sessile, lobulated polyp of the cecum, 4.2 cm in diameter, partially ulcerated. The histological examination of the biopsy revealed the presence of inflammatory granulation tissue with lymphocytic and eosinophil infiltration associated to a fibrous stroma: it was diagnosed as inflammatory fibroid polyp. Considering the polyps features (absence of a peduncle and size) that could increase the risk of a polypectomy, a surgical resection was performed. Histological examination of the specimen revealed inflammatory granulation tissue in the lamina propria, hyperplastic glands with cystic dilatations, proliferation of smooth muscle and multiple erosions on the polyp surface: this polyp was finally diagnosed as IMGP. There was also another little polyp next to the ileocecal valve, not revealed at the colonoscopy, 0.8 cm in diameter, diagnosed as tubulovillous adenoma with low grade dysplasia.ConclusionsThis is the first case of IMGP of the cecum. It is a benign lesion of unknown pathogenesis and must be considered different from other non-neoplastic polyps of the large bowel such as inflammatory cap polyps (ICP), inflammatory cloacogenic polyps, juvenile polyps (JP), inflammatory fibroid polyps (IFP), polyps secondary to mucosal prolapse syndrome (MPS), polypoid prolapsing mucosal folds of diverticular disease. When symptomatic, IMGP should be removed endoscopically, whereas surgical resection is reserved only in selected patients as in our case.

Early 'shallow' needle-knife papillotomy and guidewire cannulation: an effective and safe approach to difficult papilla.

Introduction: Needle-knife sphincterotomy (NKS), known as ‘precut’, is used worldwide to facilitate access to the common bile duct when standard cannulation has failed. This procedure is considered hazardous because it is burdened with high procedural related complications (bleeding and perforation). Its right timing is still debated. In this study we report our results using a modified precut approach, early shallow needle-knife papillotomy (eSNKP) coupled with guidewire cannulation in case of difficult papilla. We evaluated its safety and effectiveness. Methods: From 2012 to 2014, 1034 patients underwent therapeutic ERCP. A total of 138 of them presented difficult papilla and were treated with eSNKP performed after 5 failed attempts of standard guidewire cannulation. Deep biliary cannulation rate was recorded, as well as intraoperative and postoperative complication rate. Results: Successful biliary deep cannulation was achieved in 132/138 patients (95.7%) by means of eSNKP. In 6 patients (4.3%), cannulation failed even after eSNKP. ERCP was newly performed 72 hours later with successful and immediate guidewire biliary cannulation. Overall morbidity was 10.1% (14/138). No perforation occurred. Minor bleeding occurred in 4/138 cases (2.9%) and 10/138 patients (7.2%) developed mild pancreatitis. Conclusion: In case of difficult papilla, eSNKP followed by guidewire cannulation increases the successful deep biliary cannulation with low rate of complications.