Effect of Chlorine on Incorporation of Helicobacter pylori into Drinking Water Biofilms

The use of a specific peptide nucleic acid (PNA) probe demonstrated that Helicobacter pylori persisted inside biofilms exposed to low concentrations of chlorine (0.2 and 1.2 mg liter⁻¹) for at least 26 days, although no culturable cells were recovered. Coupled with data obtained using viability stains in pure culture, this result suggests that H. pylori can survive chlorination but remain undetectable by culture methods, which can be effectively replaced by PNA hybridization.Helicobacter pylori is a Gram-negative microorganism that colonizes the human stomach and can cause gastric ulcers that can degenerate into gastric carcinoma (5). The route of transmission for this pathogen is not well known, and even though culturable H. pylori has never been isolated from drinking water distribution systems (DWDS), molecular techniques such as PCR have detected the presence of H. pylori DNA in potable water (6, 15, 17), indicating that this environment could act as a reservoir for this bacterium.Chlorine is the most commonly used disinfectant worldwide to ensure the safe distribution of water to the consumer (19). Although studies conducted by Johnson et al. (14) and Baker et al. (4) have shown that H. pylori is inactivated by chlorine, their conclusions were based on the lack of recovery using standard culture plating methods which fail to consider cells that have entered a viable but nonculturable (VBNC) state. Recently, Moreno et al. (16) applied molecular techniques to demonstrate that H. pylori can survive in low concentrations of chlorine in a VBNC state. However, all these studies were performed with pure cultures using suspended cells, and until now, there have been no studies reporting on the effect of chlorination on H. pylori when associated with heterotrophic biofilms where, as is well known, microorganisms become more resistant to the biocide effect of chlorine (10).In a recent study, we demonstrated that H. pylori can be incorporated into drinking water biofilms and remain viable in the lower layers of these structures (11). It is therefore important to understand the ability of this pathogen to be incorporated and survive in heterotrophic biofilms formed in chlorinated waters. If this pathogen can remain viable under these conditions, it might therefore represent a risk to public health when released into the bulk fluid.The aim of this work was to study the effect of low concentrations of chlorine on H. pylori cells both when associated with heterotrophic biofilms and when suspended in pure culture, to assess whether the biofilm can provide protection against disinfection.     

Role of IP-10/CXCL10 in the progression of pancreatitis-like injury in mice after murine retroviral infection

Exocrinopathy and pancreatitis-like injury were developed in C57BL/6 (B6) mice infected with LP-BM5 murine leukemia virus, which is known to induce murine acquired immunodeficiency syndrome (MAIDS). The role of chemokines, especially CXCL10/interferon (IFN)-gamma-inducible protein 10 (IP-10), a chemokine to attract CXCR3+ T helper 1-type CD4+ T cells, has not been investigated thoroughly in the pathogenesis of pancreatitis. B6 mice were inoculated intraperitoneally with LP-BM5 and then injected every week with either an antibody against IP-10 or a control antibody. Eight weeks after infection, we analyzed the effect of IP-10 neutralization. Anti-IP-10 antibody treatment did not change the generalized lymphadenopathy and hepatosplenomegaly of mice with MAIDS. The treatment significantly reduced the number of IP-10- and CXCR3-positive cells in the mesenteric lymph nodes (mLNs) but not the phenotypes and gross numbers of cells. In contrast, IP-10 neutralization reduced the number of mononuclear cells infiltrating into the pancreas. Anti-IP-10 antibody treatment did not change the numbers of IFN-gamma+ and IL10+ cells in the mLN but significantly reduced their numbers, especially IFN-gamma+ and IL-10+ CD4+ T cells and IFN-gamma+ Mac-1+ cells, in the pancreas. IP-10 neutralization ameliorated the pancreatic lesions of mice with MAIDS probably by blocking the cellular infiltration of CD4+ T cells and IFN-gamma+ Mac-1+ cells into the pancreas at least at 8 wk after infection, suggesting that IP-10 and these cells might play a key role in the development of chronic autoimmune pancreatitis.     

Attenuation of mouse acute colitis by naked hepatocyte growth factor gene transfer into the liver

BACKGROUND: Hepatocyte growth factor (HGF) has multiple biological effects on a wide variety of cells. It modulates intestinal epithelial proliferation and migration, and critically regulates intestinal wound healing. AIMS: To investigate the therapeutic effect of HGF gene transfer, we introduced the HGF gene into the liver of mice with acute colitis. METHODS: The rat HGF expression plasmid vector, pCAGGS-HGF, was injected via the tail vein into C57BL/6 mice, followed by dosing with dextran sulfate sodium in distilled water. Firstly, the HGF gene was injected once on day 0. Secondly, the HGF gene was injected on day 0 and again on day 2. RESULTS: Injection of the HGF gene ameliorated colitis with inhibition of both loss of body weight and shortening of colon length. It protected the colon from epithelial erosions and cellular infiltration. Expression of mRNAs for IFN-gamma, IL18, and TNF-alpha was reduced in the colon. In contrast, expression of mRNA for IL-10 was increased. The numbers of BrdU-positive intestinal epithelial cells were increased, and the numbers of TUNEL-positive apoptotic cells were decreased. Furthermore, a second injection prolonged the elevation of serum HGF levels, and ameliorated the symptoms better than a single injection. The empty pCAGGS plasmid did not ameliorate acute colitis. CONCLUSIONS: HGF gene transfer attenuated acute colitis by facilitating intestinal wound repair as well as inhibiting inflammation, suggesting a new strategy for treatment of IBD.     

Development of an open sandwich fluoroimmunoassay based on fluorescence resonance energy transfer

We have developed a sensitive, one-step, homogeneous open sandwich fluoroimmunoassay (OsFIA) based on fluorescence resonance energy transfer (FRET) and luminescent semiconductor quantum dots (QDs). In this FRET assay, estrogen receptor beta (ER-beta) antigen was incubated with QD-labeled anti-ER-beta monoclonal antibody and Alexa Fluor (AF)-labeled anti-ER polyclonal antibody for 30 min, followed by FRET measurement. The dye separation distance was estimated between 80 and 90 A. The current method is rapid, simple, and highly sensitive, and it did not require the bound/free reagent separation steps and solid-phase carriers. A concentration as low as 0.05 nM (2.65 ng/ml) receptor was detected with linearity. In addition, the assay was performed with commercial antibodies. This assay provides a convenient alternative to conventional, laborious sandwich immunoassays.     

Anti‐fibrotic effect of chorionic plate‐derived mesenchymal stem cells isolated from human placenta in a rat model of CCl4‐injured liver: Potential application to the treatment of hepatic diseases

Translational studies have explored the therapeutic effects of stem cells, raising hopes for the treatment of numerous diseases. Here, we evaluated the therapeutic effect of chorionic plate‐derived mesenchymal stem cells (CP‐MSCs) isolated from human placenta and transplanted into rats with carbon tetrachloride (CCl₄)‐injured livers. CP‐MSCs were analyzed for hepatocyte‐specific gene expression, indocyanine green (ICG) uptake, glycogen storage, and urea production following hepatogenic differentiation. PKH26‐labeled CP‐MSCs were directly transplanted into the livers of rats that had been exposed to CCl₄ (1.6 g/kg, twice per week for 9 weeks). Blood and liver tissue were analyzed at 1, 2, and 3 weeks post‐transplantation. The expression of type I collagen (Col I) and matrix metalloproteinases (MMPs) was analyzed in rat T‐HSC/Cl‐6 hepatic stellate cells co‐cultured with CP‐MSCs following exposure to TGF‐β. The expression levels of α‐smooth muscle actin (α‐SMA) and Col I were lower in transplanted (TP) rats than in non‐transplanted (Non‐TP) animals (P < 0.05), whereas the expression levels of albumin and MMP‐9 were increased. TP rats exhibited significantly higher uptake/excretion of ICG than non‐TP rats (P < 0.005). In addition, collagen synthesis in T‐HSC/Cl‐6 cells exposed to TGF‐β was decreased by co‐culture with CP‐MSCs, which triggered the activation of MMP‐2 and MMP‐9. These results contribute to our understanding of the potential pathophysiological roles of CP‐MSCs, including anti‐fibrotic effects in liver disease, and provide a foundation for the development of new cell therapy‐based strategies for the treatment of difficult‐to‐treat liver diseases. J. Cell. Biochem. 111: 1453–1463, 2010. © 2010 Wiley‐Liss, Inc.     

Effects of initial density,nutrient, and water level regime on the seedling survival and growth of <Emphasis Type="Italic">Typha orientalis</Emphasis> Presl

Seedlings’ successful establishment is of importance in the preemption process of pioneers in wetlands. Although Typha orientalis Presl has been reported as a pioneer in Asia countries, studies on the seedling phase of T. orientalis are not available yet. A mesocosm experiment was conducted to understand the effects of biotic (initial density) and abiotic (nutrient and water level regime) factors on the seedling survival and growth of T. orientalis. Most seedlings survived under low initial density (93.8%) and eutrophic (95.5%) rather than high initial density (64.3%) and ombrotrophic (62.5%). Seedlings under low initial density, eutrophic, and flooded conditions showed relatively higher growth in shoot height. The final number of ramets showed an adverse tendency compared to the survival rate and shoot height particularly depending on the water level regime. T. orientalis compensated its biomass production with producing less but longer shoots under the flooded condition, indicating the phenotypic plasticity of T. orientalis as a deep water species. However, the compensation seemed to be guaranteed only under the condition of sufficient nutrients. Asian T. orientalis seemed not to be a pioneer but a weak-competitor not only in mature plant stage but also in juvenile seedling stage unless sufficient nutrients are guaranteed.