Abstract The intestinal epithelium serves as a barrier to the flora. In response pathogens, epithelial cells (IEC) secrete proinflammatory cytokines. To aid in defense against bacteria, IEC also antimicrobial peptides, termed defensins. aim of our studies was understand role TLR signaling regulation β-defensin expression by IEC. effect LPS and peptidoglycan on β-defensin-2 examined lines constitutively or transgenically expressing TLRs. Regulation assessed using promoter-reporter constructs...

Monodispersed dendritic poly(L-lysine)s (DPKs) of several generations were synthesized, and their characteristics as a gene transfection reagent then investigated. The agarose gel shift ethidium bromide titration assay proved that the DPKs third generation higher could form complex with plasmid DNA, degree compaction DNA was increased by increasing number generation. fifth sixth generation, which have 64 128 amine groups on surface molecule, respectively, showed efficient ability into...

The decrease in overall translation activity occurring concomitantly with the transition from exponential growth phase to stationary of Escherichia coli cells was found be accompanied by appearance 100S ribosomes (dimers 70S ribosome monomers). Analysis ribosomal proteins radical-free and highly reducing method two-dimensional gel electrophoresis indicated that a protein, designated protein E, exclusively associated ribosomes. From results, we propose E is "ribosome modulation factor" (RMF),...

<i>Helicobacter pylori</i>, a Gram-negative gastric bacterium, secretes VacA, cytotoxin that causes vacuolar degeneration of susceptible cells. Velocity sedimentation analysis showed treatment VacA at alkaline pH led to disassembly oligomers, an observation reported previously for acid-treated VacA. Exposure acid or alkali increased its binding AZ-521 cells, as shown by indirect immunofluorescence and flow cytometry. Moreover, immunoprecipitates with polyclonal antibodies against from cells...

Abstract Mucosal mast cells strategically located at the optimal site interact with invading bacteria. Presence of VacA, virulent Helicobacter pylori cytotoxin, is correlated severity H. pylori-induced gastritis. To examine mechanisms inflammation in gastritis, we administered VacA to mice. Inoculation resulted epithelium vacuolization and marked infiltrations mononuclear into mucosal within 24 h. In an vitro study using bone marrow-derived cells, directly bound showed a chemotactic activity...

Polycationic reagents such as cationic lipids and poly-l-lysine are widely used for gene transfer into cells <i>in vitro</i> show promise vectors vivo</i> therapy applications nonviral techniques. We have developed a novel transfection method using amphiphilic α-helical oligopeptides with repeated sequences. Oligopeptide has the advantages of being easily designed modified because its simple structure. In this study, we synthesized five kinds peptides which total chain length width...

Primary cultures of guinea pig gastric mucosal cells express NADPH oxidase 1 (Nox1), a homolog gp91(phox), and produce superoxide anion (O2-) at rate approximately 100 nmol.mg protein(-1).h(-1) in response to Helicobacter pylori (H. pylori) lipopolysaccharide (LPS) from virulent type I strains. The upregulated O2- production also enhances H. LPS-stimulated tumor necrosis factor-alpha or cyclooxygenase-2 mRNA expression, which suggests potential role for Nox1 the pathogenesis...

Antimicrobial peptides are crucial for host defense at mucosal surfaces. Bacterial factors responsible induction of human beta-defensin-2 (hBD-2) mRNA expression in Caco-2 carcinoma cells were determined. Salmonella enteritidis, typhimurium, typhi, dublin, and culture supernatants these strains induced hBD-2 cells. Using luciferase as a reporter gene approximately 2.1-kilobase pair promoter, the hBD-2-inducing factor supernatant S. enteritidis was isolated. The heat-stable...

Chronic infection with cagA -positive Helicobacter pylori is associated the development of atrophic gastritis, peptic ulcers, and gastric adenocarcinoma. The gene product CagA injected into epithelial cells, where it undergoes tyrosine phosphorylation by Src family kinases. Translocated disturbs cellular functions physically interacting deregulating intracellular signaling transducers through both phosphorylation-dependent -independent mechanisms. To gain further insights pathophysiological...

In human airways, epithelial cells lining the lumen and intraluminal (e.g., polymorphonuclear cells) participate in innate immune response. These secrete or express on their surfaces arginine-specific ADP ribosyltransferases. Defensins, antimicrobial proteins secreted by cells, are arginine-rich, leading us to hypothesize that ribosylation could modify biological activities. We found an ribosyltransferase-1 present airway modifies Arg-14 of alpha defensin-1. ADP-ribosylated defensin-1 had...

Helicobacter pylori vacuolating cytotoxin, VacA, induces vacuolation, mitochondrial damage, cytochrome c release, and apoptosis of gastric epithelial cells. To detect proteins that serve as VacA receptors, we used co-immunoprecipitation techniques following biotinylation the cell surface identified p250, a receptor-like protein-tyrosine phosphatase β (RPTPβ) VacA-binding protein (Yahiro, K., Niidome, T., Kimura, M., Hatakeyama, Aoyagi, H., Kurazono, Imagawa, Wada, A., Moss, J., Hirayama, T....

Persistent Helicobacter pylori colonization in the stomach induces gastritis and peptic ulcer interferes with healing. Most strains of H. produce a cytotoxin, VacA, that cytoplasmic vacuolation epithelial cells structural functional changes, leading to gastric injury. VacA is known cause cell death by mitochondrial damage. We hypothesized might disrupt other signaling pathways; end, we examined effects on MAPKs elucidate their role abnormalities seen VacA-treated cells. stimulated...

The epidemiology of enteric pathogens has not been well studied in Kenya because wide disparities health status across the country. Therefore, present study describes prevalence enteropathogenic bacteria, their seasonal variation, and antibiotic resistance profiles among hospitalized diarrheic children a suburban region central Kenya.Fecal samples were collected between July 2009 December 2013 from total 1410 younger than 5 years, with acute diarrhea Kiambu County Hospital, Kenya....

Human beta-defensins (hBDs) are important antimicrobial peptides that contribute to innate immunity at mucosal surfaces. This study was undertaken investigate the expression of hBD-1 and hBD-2 in intrahepatic biliary epithelial cells specimens human liver, 4 cultured cell lines (2 consisting 2 cholangiocarcinoma cells). In addition, were assayed bile. nonspecifically expressed immunohistochemically epithelium hepatocytes all patients studied, but restricted large bile ducts 8 10 with...

Human β-defensin 2 (hBD-2) is an antimicrobial peptide involved in host defence against bacterial infection epithelial tissues. Its levels are dramatically increased after infection. The involvement of NF-κB Helicobacter pylori-mediated induction hBD-2 promoter activity was examined. A luciferase reporter plasmid containing the extending from −2110 base pairs to −1 transiently expressed MKN45 cells, and determined incubation with H. pylori for 6 h. Deletion or mutation site at −208 abolished...

Background Information is limited about how adipokines predict the accumulation of cardiovascular (CV) risk factors or presence metabolic syndrome (MS) in children. Methods and Results The subjects were 321 children (200 boys 121 girls; 109 normal 212 obese) aged 6-12 years. Obesity was defined as a body mass index ≥ 95th percentile for age sex. MS by using newly established Task Force criteria. levels - adiponectin, leptin, ghrelin, high sensitive C-reactive protein (CRP) resistin -were...

Phorbol 12-myristate 13-acetate (PMA) induces differentiation of human leukemic HL-60 cells into with macrophage-like characteristics and enhances the susceptibility to <i>Helicobacter pylori</i> VacA toxin (de Bernard, M., Moschioni., Papini, E., Telford, J. L., Rappuoli, R., Montecucco, C. (1998) <i>FEBS Lett.</i> 436, 218–222). We examined mechanism by which acquire sensitivity VacA, in particular, looking for expression RPTPβ, a VacA-binding protein postulated be receptor (Yahiro, K.,...

Helicobacter pylori vacuolating cytotoxin, VacA, induces multiple effects on epithelial cells through different cellular events: one involves pore formation, leading to vacuolation, mitochondrial damage, and apoptosis, the second cell signaling, resulting in stimulation of proinflammatory responses detachment. Our recent data demonstrated that VacA uses receptor-like protein tyrosine phosphatase beta (RPTPbeta) as a receptor, which five residues (QTTQP) at positions 747 751 are involved...

Antimicrobial peptides such as defensins are crucial for host defense at mucosal surfaces. We reported previously that Salmonella enteritidis flagellin (FliC) induced human β-defensin-2 (hBD-2) mRNA expression in Caco-2 cells via NF-κB activation (Ogushi, K., Wada, A., Niidome, T., Mori, N., Oishi, Nagatake, Takahashi, Asakura, H., Makino, S., Hojo, Nakahara, Y., Ohsaki, M., Hatakeyama, Aoyagi, Kurazono, Moss, J., and Hirayama, T. (2001) J. Biol. Chem. 276, 30521–30526). In this study, we...

Defensins are small cationic bactericidal peptides present abundantly in the granules of polymorphonuclear neutrophils (PMNs). Human PMNs contain four defensins termed HNP-1 to HNP-4. We used a new assay system agar plates, radial diffusion assay, evaluate effects human against gram-positive and -negative bacteria. A crude mixture HNP-1, -2, -3 (crude HNPs) was purified from PMN extracts by reversed-phase high-pressure liquid chromatography (RP-HPLC). The different components were later...