Pectic oligosaccharide structure-function relationships: prebiotics, inhibitors of Escherichia coli O157:H7 adhesion and reduction of Shiga toxin cytotoxicity in HT29 cells

Shiga toxin (Stx)-producing, food-contaminating Escherichia coli (STEC) is a major health concern. Plant-derived pectin and pectic-oligosaccharides (POS) have been considered as prebiotics and for the protection of humans from Stx. Of five structurally different citrus pectic samples, POS1, POS2 and modified citrus pectin 1 (MCP1) were bifidogenic with similar fermentabilities in human faecal cultures and arabinose-rich POS2 had the greatest prebiotic potential. Pectic oligosaccharides also enhanced lactobacilli growth during mixed batch faecal fermentation. We demonstrated that all pectic substrates were anti-adhesive for E. coli O157:H7 binding to human HT29 cells. Lower molecular weight and deesterification enhanced the anti-adhesive activity. We showed that all pectic samples reduced Stx2 cytotoxicity in HT29 cells, as measured by the reduction of human rRNA depurination detected by our novel TaqMan-based RT-qPCR assay, with POS1 performing the best. POS1 competes with Stx2 binding to the Gb3 receptor based on ELISA results, underlining the POS anti-STEC properties

Activation of Human T-Helper/Inducer Cell, T-Cytotoxic Cell, B-Cell, and Natural Killer (NK)-Cells and induction of Natural Killer Cell Activity against K562 Chronic Myeloid Leukemia Cells with Modified Citrus Pectin

<p>Abstract</p> <p>Background</p> <p>Modified citrus pectin (MCP) is known for its anti-cancer effects and its ability to be absorbed and circulated in the human body. In this report we tested the ability of MCP to induce the activation of human blood lymphocyte subsets like T, B and NK-cells.</p> <p>Methods</p> <p>MCP treated human blood samples were incubated with specific antibody combinations and analyzed in a flow cytometer using a 3-color protocol. To test functionality of the activated NK-cells, isolated normal lymphocytes were treated with increasing concentrations of MCP. Log-phase PKH26-labeled K562 leukemic cells were added to the lymphocytes and incubated for 4 h. The mixture was stained with FITC-labeled active form of caspase 3 antibody and analyzed by a 2-color flow cytometry protocol. The percentage of K562 cells positive for PKH26 and FITC were calculated as the dead cells induced by NK-cells. Monosaccharide analysis of the MCP was performed by high-performance anion-exchange chromatography with pulse amperometric detection (HPAEC-PAD).</p> <p>Results</p> <p>MCP activated T-cytotoxic cells and B-cell in a dose-dependent manner, and induced significant dose-dependent activation of NK-cells. MCP-activated NK-cells demonstrated functionality in inducing cancer cell death. MCP consisted of oligogalacturonic acids with some containing 4,5-unsaturated non-reducing ends.</p> <p>Conclusions</p> <p>MCP has immunostimulatory properties in human blood samples, including the activation of functional NK cells against K562 leukemic cells in culture. Unsaturated oligogalacturonic acids appear to be the immunostimulatory carbohydrates in MCP.</p

Identification of Lactobacillus Strains Capable of Fermenting Fructo-Oligosaccharides and Inulin

Novel probiotic strains that can ferment prebiotics are important for functional foods. The utilization of prebiotics is strain specific, so we screened 86 Lactobacillus strains and compared them to Bifidobacterium breve 2141 for the ability to grow and produce SCFA when 1% inulin or fructo-oligosaccharides (FOS) were provided as the carbon source in batch fermentations. When grown anaerobically at 32 °C, ten Lactobacillus strains grew on both prebiotic substrates (OD600 ≥ 1.2); while Lactobacillus coryniformis subsp. torquens B4390 grew only in the presence of inulin. When the growth temperature was increased to 37 °C to simulate the human body temperature, four of these strains were no longer able to grow on either prebiotic. Additionally, L. casei strains 4646 and B441, and L. helveticus strains B1842 and B1929 did not require anaerobic conditions for growth on both prebiotics. Short-chain fatty acid analysis was performed on cell-free supernatants. The concentration of lactic acid produced by the ten Lactobacillus strains in the presence of prebiotics ranged from 73–205 mM. L. helveticus B1929 produced the highest concentration of acetic acid ~19 mM, while L. paraplantarum B23115 and L. paracasei ssp. paracasei B4564 produced the highest concentrations of propionic (1.8–4.0 mM) and butyric (0.9 and 1.1 mM) acids from prebiotic fermentation. L. mali B4563, L. paraplantarum B23115 and L. paracasei ssp. paracasei B4564 were identified as butyrate producers for the first time. These strains hold potential as synbiotics with FOS or inulin in the development of functional foods, including infant formula

Modified citrus pectin confers a preventative effect on cancer-related pathways in CdCl2-treated C. elegans

Cadmium chloride (CdCl2) is a toxic compound found as a pollutant in the environment due to agricultural and industrial sources. Exposure to Cd2+ is known to promote malignant tumors such as lung cancer and leukemia. While the current medications for cadmium toxicity focus on treatments to promote the excretion from the body, treatments to improve health after cadmium exposure are less well studied. Modified citrus pectin (MCP) is a polysaccharide derived from citrus peels that has been shown to induce natural killer cell activity in myeloid leukemia cells and also act as a natural chelation agent to help excrete toxic metals from healthy human subjects. We hypothesized that MCP might have a counteracting effect against CdCl2 toxicity through cancer-related pathways. This study investigates the effects of MCP on CdCl2 toxicity in C. elegans, which shares a number of cancer-related pathways with mammals. The results indicated that MCP was able to significantly counter the toxic effects of CdCl2 on C. elegans lifespan and development. Our studies suggest that the beneficial effects of MCP may result from its ability to mitigate the effects of CdCl2 on gene expression, particularly in conserved pathways associated with apoptosis, tumor induction and suppression and inflammation-related pathways