Effect of Digested Selected Food Items on Markers of Oxidative Stress and Inflammation in a Caco-2-Based Human Gut Epithelial Model.

The human gut epithelium presents a crucial interface between ingested food items and the host. Understanding how different food items influence oxidative stress and inflammation in the gut is of great importance. This study assessed the impact of various digested food items on oxidative stress, inflammation, and DNA/RNA damage in human gut epithelial cells. Differentiated Caco-2 cells were exposed to food items and their combinations (n = 22) selected from a previous study, including sausage, white chocolate, soda, coffee, orange juice, and curcumin. Following stimulation with TNF-α/IFN-1ß/LPS and H2O2 for 4 h, the cells were exposed to digested food items or appropriate controls (empty digesta and medium) for a further 16 h. Cell viability, antioxidant capacity (ABTS, FRAP), IL-6, IL-8, F2-isoprostanes, lipid peroxidation (MDA), and DNA/RNA oxidative damage were assessed (3 independent triplicates). The ABTS assay revealed that cells treated with "white chocolate" and "sausage + coffee" exhibited significantly reduced antioxidant capacity compared to stimulated control cells (ABTS = 52.3%, 54.8%, respectively, p < 0.05). Similar results were observed for FRAP (sausage = 34.9%; white chocolate + sausage = 35.1%). IL-6 levels increased in cells treated with "white chocolate + sausage" digesta (by 101%, p < 0.05). Moreover, MDA levels were significantly elevated in cells treated with digested "sausage" or sausage in combination with other food items. DNA/RNA oxidative damage was found to be higher in digesta containing sausage or white chocolate (up to 550%, p < 0.05) compared to stimulated control cells. This investigation provides insights into how different food items may affect gut health and underscores the complex interplay between food components and the epithelium at this critical interface of absorption.

Pro- and Antioxidant Effect of Food Items and Matrices during Simulated In Vitro Digestion.

The digestive tract can be considered a bioreactor. High levels of reactive oxygen species (ROS) during digestion may predispose for local and/or systemic oxidative stress and inflammation, e.g., inflammatory bowel diseases. Food items rich in antioxidants may prevent such aggravation. This investigation analyzed pro-and antioxidant patterns of food matrices/items following in vitro digestion. Gastrointestinal digestion reflecting typically consumed quantities was performed on nine food items (orange and tomato juice, soda, coffee, white chocolate, sausage, vitamin C and E, and curcumin) and their combinations (n = 24), using the INFOGEST model. Antioxidant potential was measured by FRAP, DPPH, and ABTS, and pro-oxidant aspects by MDA (malondialdehyde) and peroxide formation. An anti-pro-oxidant score was developed, combining the five assays. Liquid food items showed moderately high antioxidant values, except for coffee and orange juice, which exhibited a high antioxidant potential. Solid matrices, e.g., white chocolate and sausage, showed both high pro-oxidant (up to 22 mg/L MDA) and high antioxidant potential (up to 336 mg/L vitamin C equivalents) at the same time. Individual vitamins (C and E) at physiological levels (achievable from food items) showed a moderate antioxidant potential (<220 mg/L vitamin C equivalents). Overall, both antioxidant and pro-oxidant assays correlated well, with correlation coefficients of up to 0.894. The effects of food combinations were generally additive, i.e., non-synergistic, except for combinations with sausage, where strong quenching effects for MDA were observed, e.g., with orange juice. In conclusion, as especially highlighted by complex matrices demonstrating both pro- and antioxidant potential, only measuring one aspect would result in physiological misinterpretations. Therefore, it is imperative to employ a combination of assays to evaluate both pro- and antioxidant properties of food digesta to ensure physiological relevance.

Potential health effects of brewers' spent grain as a functional food ingredient assessed by markers of oxidative stress and inflammation following gastro-intestinal digestion and in a cell model of the small intestine.

Brewer's spent grains (BSG) are a by-product of the beer-brewing industry, often employed as animal feeding stuffs. With BSG being rich not only in proteins, lipids, and dietary fiber but also in certain phytochemicals, it constitutes a potentially valuable food source that could be employed as a functional food, e.g. against chronic inflammatory diseases. Several types of bread were prepared with various amounts of BSG as flour replacement (0, 10, 20, 40, 60, 80 and 100%), either employing wet BSG or dried BSG after pressing. Total phenolics, flavonoids, insoluble dietary fiber, as well as antioxidant capacity (FRAP, ABTS) were measured in the bread, before and after simulated gastro-intestinal digestion. Furthermore, we investigated digested BSG and bread-containing BSG for their capability to alter oxidative stress (Nrf2, malondialdehyde) and inflammation (IL-6, IL-8, NO, and PGE2) in a Caco-2 cell culture model of the small intestine. Incorporation of BSG significantly and dose-dependently enhanced the amount of dietary fiber in the product, as well as total phenolics, flavonoids, and antioxidant capacity, by over 10-fold, 3-fold, 4-fold and 5-fold, respectively, when replacing all of the flour with BSG. This pattern remained after in vitro digestion. However, digesta failed to show significant antioxidant or anti-inflammatory effects on the biomarkers observed in the cell model. Consuming 150 g of such a BSG-bread (wet based) would supply the proposed RDA of 25 g d-1 dietary fiber and could be a healthy product valorizing BSG.

Determination of genes and microRNAs involved in the resistance to fludarabine in vivo in chronic lymphocytic leukemia.

BACKGROUND: Chronic lymphocytic leukemia (CLL) cells are often affected by genomic aberrations targeting key regulatory genes. Although fludarabine is the standard first line therapy to treat CLL, only few data are available about the resistance of B cells to this purine nucleoside analog in vivo. Here we sought to increase our understanding of fludarabine action and describe the mechanisms leading to resistance in vivo. We performed an analysis of genomic aberrations, gene expression profiles, and microRNAs expression in CLL blood B lymphocytes isolated during the course of patients' treatment with fludarabine. RESULTS: In sensitive patients, the differentially expressed genes we identified were mainly involved in p53 signaling, DNA damage response, cell cycle and cell death. In resistant patients, uncommon genomic abnormalities were observed and the resistance toward fludarabine could be characterized based on the expression profiles of genes implicated in lymphocyte proliferation, DNA repair, and cell growth and survival. Of particular interest in some patients was the amplification of MYC (8q) observed both at the gene and transcript levels, together with alterations of myc-transcriptional targets, including genes and miRNAs involved in the regulation of cell cycle and proliferation. Differential expression of the sulfatase SULF2 and of miR-29a, -181a, and -221 was also observed between resistant and sensitive patients before treatment. These observations were further confirmed on a validation cohort of CLL patients treated with fludarabine in vitro. CONCLUSION: In the present study we identified genes and miRNAs that may predict clinical resistance of CLL to fludarabine, and describe an interesting oncogenic mechanism in CLL patients resistant to fludarabine by which the complete MYC-specific regulatory network was altered (DNA and RNA levels, and transcriptional targets). These results should prove useful for understanding and overcoming refractoriness to fludarabine and also for predicting the clinical outcome of CLL patients before or early during their treatment.

Valproic acid induces non-apoptotic cell death mechanisms in multiple myeloma cell lines.

Multiple myeloma (MM) is an incurable hematological disorder characterized by dysregulated proliferation of terminally differentiated plasma cells. Aberrant histone acetylation has been observed in the development of numerous malignancies. Histone deacetylase inhibitors such as valproic acid (VPA) are promising drugs for cancer therapy since they have been reported to have antiproliferative effects and to induce differentiation in carcinoma and leukemic cells. Considering the advantage of being already in clinical use for epilepsy treatment, valproic acid might be a promising therapeutic candidate drug in the management of multiple myeloma. In this study, we show that the short fatty acid VPA has a time and dose-dependent cytotoxic effect on the MM cell lines OPM2, RPMI and U266. The influence of VPA on cell cycle and apoptosis have been evaluated by flow cytometry. Our results show that the three cell lines are blocked in G0/G1 phase. The observed sensitivity to VPA can be partially explained by late apoptosis. Since caspase 3 is activated in all tested cell lines after VPA treatment, a caspase-dependent pathway seems to be involved but not activated by the classic apoptotic pathways. We have also studied another mechanism of cell death, the senescence-like phenotype, but did not find any evidence for its implication. Thus, treatment with VPA may imply other alternative cell death mechanisms.