Enteric Toxins from Bacteria Colonizing Human Gut

The large and heterogeneous microbial population colonising the human intestinal tract includes a number of aerobic and anaerobic bacteria that produce one or more toxins. While exhibiting very different physico-chemical properties these exotoxins share the ability to penetrate intestinal cells after their binding to a specific surface receptor, thus reaching a subcellular target at membrane or cytoskeleton level. The most relevant in vitro and in vivo data, reported in the literature, on the mode of action of the major enterotoxins and cytotoxins produced by bacteria belonging to the human gut microora are reviewed in the light of our recent knowledge on bacteria-host cell interactions

A Scanning Electron Microscopy Analysis of Human Cytotoxic Cell Subsets and of Their Mode of Conjugation with Tumor Cell Targets

In this study we describe the surface features of non-MHC (Major Histocompatibility Complex)-restricted cytotoxic cells isolated from human peripheral blood. Purified populations of CD3-, natural killer (NK) cells were allowed to interact with NK-sensitive (K562) and NK-resistant (THP-1-0) tumor cell targets. The type of effector to target cell binding was investigated by scanning electron microscope (SEM) analysis. A different interaction with the effectors is described for NK-resistant targets in comparison with NK-susceptible tumor cells. SEM was also used to investigate the relationship between interleukin 2 (IL2)-activated cytotoxic cells (lymphokine-activated killer, LAK, cells) and the tumor targets. We also describe the unique growth features of certain clones of cytotoxic T cells expressing γδ antigen receptors which support the contention that these cells may have a special ability of homing into tissues. We conclude that non-specific cytotoxic cells constitute a diverse population of effectors which differ not only for the expression of surface antigens, but also for their ability to interact with tumor cell targets and to home into the peripheral tissues where they may exert their lytic functions

A microbiological and morphological study of blocked biliary stents

Biliary stent blockage represents the main limitation of the use of such devices in relieving obstructive jaundice due to a variety of malignant and benign conditions. Microbiological and morphological analysis of the occluding sludge present on the inner surface of 30 biliary stents was performed to evaluate the different components of such material and the effect of the antibiotic treatment on the biofilm formation. A highly organized biofilm, constituted by microbial cells embedded in an amorphous matrix together with crystallized bile salts, was observed Enterococcus spp . represented the most common isolate from both occluded and non-occluded stents. The antibiotic therapy, while selecting for multi-resistant bacteria and fungi, might possibly delay the biofilm formation. Key words: biliary stents, microbial colonization, biofilm

Escherichia coli Cytotoxic Necrotizing Factor 1 (CNF1), a Toxin That Activates the Rho GTPase

Cytotoxic necrotizing factor 1 (CNF1), a 110-kDa protein toxin from pathogenic Escherichia coli induces actin reorganization into stress fibers and retraction fibers in human epithelial cultured cells allowing them to spread. CNF1 is acting in the cytosol since microinjection of the toxin into HEp-2 cells mimics the effects of the externally applied CNF1. Incubation in vitro of CNF1 with recombinant small GTPases induces a modification of Rho (but not of Rac, Cdc42, Ras, or Rab6) as demonstrated by a discrete increase in the apparent molecular weight of the molecule. Preincubation of cells with CNF1 impairs the cytotoxic effects of Clostridium difficile toxin B, which inactivates Rho but not those of Clostridium sordellii LT toxin, which inhibits Ras and Rac. As shown for Rho-GTP, CNF1 activates, in a time- and dose-dependent manner, a cytoskeleton-associated phosphatidylinositol 4-phosphate 5-kinase. However, neither the phosphatidylinositol 4,5-bisphosphate (PIP2) nor the phosphatidylinositol 3,4-bisphosphate (PI 3,4-P2) or 3,4,5-trisphosphate (PIP3) cellular content were found increased in CNF1 treated HEp-2 cells. Cellular effects of CNF1 were not blocked by LY294002, a stable inhibitor of the phosphoinositide 3-kinase. Incubation of HEp-2 cells with CNF1 induces relocalization of myosin 2 in stress fibers but not in retraction fibers. Altogether, our data indicate that CNF1 is a toxin that selectively activates the Rho GTP-binding protein, thus inducing contractility and cell spreading

Aeromonas sobria sepsis complicated by rhabdomyolysis in an HIV-positive patient: case report and evaluation of traits associated with bacterial virulence

Summary Human infection with Aeromonas species is uncommon and most often due to trauma with exposure to contaminated water or soil. A 43-year-old HIV- and hepatitis C virus (HCV)-infected male, after a two-week course of corticosteroid therapy for an autoimmune anemia, developed diarrhea, dermatologic manifestations and a multiple organ dysfunction syndrome, resulting in death. Although stool samples were repeatedly negative, two sets of blood cultures obtained during a single peak of fever yielded the post-mortem isolation of a Gram-negative, oxidase-positive, β-hemolytic bacillus that was identified as Aeromonas sobria . Empiric antibiotic therapy was unsuccessful. Evaluation of the virulence-associated traits of the clinical isolate (adhesion, cytotoxicity activity, biofilm production) showed that the strain was a poor producer of recognized virulence factors, thereby indicating that the unfortunate coexistence of HIV infection, HCV-related liver cirrhosis and corticosteroids played a key role in the clinical course

Role of multispecies microbial biofilms in the occlusion of biliary stents

Endoscopic stenting is a standard palliative approach for the treatment of a variety of diseases involving biliary obstruction. However, the major limitation of this approach is represented by stent occlusion followed by life-threatening cholangitis, often requiring stent removal and replacement with a new one. Although it is generally believed that microbial colonization of the inner surface of the stent plays an important role in initiating the clogging process, so far available data are not enough for a full understanding of this phenomenon. In fact, it is known that when a biliary stent is inserted across the sphincter of Oddi, the loss of the antimicrobial barrier represented by the sphincter itself and the low pressure in the common bile duct allow reflux of duodenal content, thus promoting an ascending microbial colonization. The sessile mode of growth and the exopolysaccharide production, which leads to the subsequent establishment of a thick biofilm, provides microorganisms with an efficient protection from both antibacterial agents and phagocytic cells. The aim of this study was to analyze the tridimensional structure of the microbial biofilm grown in the lumen of 15 clogged biliary stents and to identify the microbial species involved in the clogging process. Scanning electron microscopy investigations revealed that sludge present in the stent lumen consist of a rich and assorted microbial flora, including aerobic and anaerobic species, mixed with a large amount of amorphous material containing dietary fibres, crystals of cholesterol and other precipitates of bacteria-driven bile salts. Key words: biliary stent, biofilm, microbial colonization, stent occlusio

Comparison of triple-lumen central venous catheters impregnated with silver nanoparticles (AgTive®) vs conventional catheters in intensive care unit patients

BACKGROUND: Silver-impregnated central venous catheters (CVCs) have been proposed as a means for preventing CVC colonization and related bloodstream infections (CRBSIs). AIM: To evaluate the efficacy of CVCs impregnated with silver nanoparticles in a large group of critically ill patients. METHODS: A prospective, randomized clinical trial was conducted in five intensive care units (ICUs). Three hundred and thirty-eight adult patients requiring CVCs between April 2006 and November 2008 were randomized to receive AgTive silver-nanoparticle-impregnated (SC) or conventional (CC) CVCs. Primary endpoints were CVC colonization (growth of ≥15 colony-forming units from the catheter tip) and incident CRBSIs (meeting the definitions of the Centers for Disease Control and Prevention). Infection-free time (days from initial CVC insertion to initial blood culture positivity) and ICU mortality rates were measured as secondary endpoints. FINDINGS: The SC group (N = 135) and CC group (N = 137) were similar in terms of clinical and laboratory parameters at baseline, reasons for ICU admission, complications during CVC insertion, and total time with CVC (mean ± standard deviation; SC 13 ± 24 vs CC 15 ± 37 days). No significant intergroup differences were found in CVC colonization rates (SC 32.6% vs CC 30%; P = 0.7), CRBSI incidence rates (3.36 infections per 1000 catheter-days in both groups), infection-free times (SC 13 ± 34 vs CC 12 ± 12 days; P = 0.85) or ICU mortality (SC 46% vs CC 43%; P = 0.7). CONCLUSION: In critically ill patients, use of AgTive(®) silver-nanoparticle-impregnated CVCs had no significant effect on CVC colonization, CRBSI incidence or ICU mortality. These CVCs cannot be recommended as an adjunctive tool for control of CRBSIs

Biofilm-Based Nosocomial Infections

Microbial biofilms have been implicated in a large number of acute and chronic infections, as well as in the failure of antibiotic treatment, particularly in hospitalized patients. In fact, the well-known persistence in the nosocomial environment of multidrug resistant microorganisms is believed to be highly promoted by the ability of the great majority of the involved bacterial and fungal species to adhere on living or abiotic surfaces, and to grow in sessile mode, to form single- or multi-species biofilms. In these communities, microbes grow encased in a hydrated matrix of extracellular polymeric substances produced by themselves and are well protected from the host immune response and the attack of antimicrobial molecules. Thus, the establishment of microbial biofilm communities on the mucosal and soft tissues of hospitalized patients, as well as on the surfaces of indwelling devices and medical instruments, is expected to have a great influence on the success of the antibiotic therapies against most of the bugs involved in nosocomial infections, being biofilm-growing bacteria and fungi much less susceptible to antibiotics