scholarly journals Evaluation of the Individual and Combined Toxicity of Fumonisin Mycotoxins in Human Gastric Epithelial Cells

2020 ◽  
Vol 21 (16) ◽  
pp. 5917
Author(s):  
Song Yu ◽  
Bingxuan Jia ◽  
Na Liu ◽  
Dianzhen Yu ◽  
Aibo Wu
Keyword(s):  
Synergistic Effect ◽  
Fumonisin B1 ◽  
Potential Method ◽  
Assessment Method ◽  
Antagonistic Effect ◽  
Epithelial Cell Line ◽  
High Concentration ◽  
Food And Feed ◽  
The Individual

Fumonisin contaminates food and feed extensively throughout the world, causing chronic and acute toxicity in human and animals. Currently, studies on the toxicology of fumonisins mainly focus on fumonisin B1 (FB1). Considering that FB1, fumonisin B2 (FB2) and fumonisin B3 (FB3) could coexist in food and feed, a study regarding a single toxin, FB1, may not completely reflect the toxicity of fumonisin. The gastrointestinal tract is usually exposed to these dietary toxins. In our study, the human gastric epithelial cell line (GES-1) was used as in vitro model to evaluate the toxicity of fumonisin. Firstly, we found that they could cause a decrease in cell viability, and increase in membrane leakage, cell death and the induction of expression of markers for endoplasmic reticulum (ER) stress. Their toxicity potency rank is FB1 > FB2 >> FB3. The results also showed that the synergistic effect appeared in the combinations of FB1 + FB2 and FB1 + FB3. Nevertheless, the combinations of FB2 + FB3 and FB1 + FB2 + FB3 showed a synergistic effect at low concentration and an antagonistic effect at high concentration. We also found that myriocin (ISP-1) could alleviate the cytotoxicity induced by fumonisin in GES-1 cells. Finally, this study may help to determine or optimize the legal limits and risk assessment method of mycotoxins in food and feed and provide a potential method to block the fumonisin toxicity.

scholarly journals In vitro antagonistic effect of nisin on faecal enterococci and staphylococci

2001 ◽  
Vol 46 (No. 9–10) ◽  
pp. 237-240 ◽  
Author(s):  
A. Lauková ◽  
I. Štyriak ◽  
M. Mareková
Keyword(s):  
Broad Spectrum ◽  
Antagonistic Effect ◽  
Individual Species ◽  
European Bison ◽  
Nisin Concentration ◽  
Faecal Samples ◽  
The Individual

Enterococci and staphylococci, isolates from faecal samples of 46 different animals such as deer, chamois, European bison, zebra, camel, antelope, gazelle, horse, and piglets were treated by nisin (concentration 1 mg/ml). Only two strains (SX38 and EA163), isolates from the faeces of deer were not inhibited by nisin under in vitro conditions. It means 97.4% of target isolates were inhibited by nisin and 2.6% were resistant. The majority of microorganisms were inhibited by nisin under MIC 1.56–100 µg/ml. Twenty-two percent out of 77 isolates were inhibited by MIC of nisin 3.12 µg.Enterococcus sp. E6B strain was found the most sensitive (inhibition by MIC 1.56 µg of nisin). Although only a few staphylococci were tested, most of them were inhibited by nisin. Even though the effect of nisin on the individual species was not evaluated, its effect on the group of bacteria is already important. In general, the properties of nisin indicate a broad spectrum of its utilization.

scholarly journals A264 HELMINTH-INFECTION MOBILIZES HOST AND MICROBIAL FACTORS THAT CO-OPERATE TO SUPPRESS CHEMICAL-INDUCED COLITIS IN MICE

2020 ◽  
Vol 3 (Supplement_1) ◽  
pp. 141-142
Author(s):  
A J Shute ◽  
B E Callejas Pina ◽  
T S Jayme ◽  
A Wang ◽  
A Buret ◽  
...  
Keyword(s):  
Drinking Water ◽  
Murine Model ◽  
Hymenolepis Diminuta ◽  
Model Systems ◽  
Epithelial Cell Line ◽  
Helminth Parasites ◽  
Critical Determinant ◽  
The Individual ◽  
Helminth Therapy

Abstract Background Infection with helminth parasites suppresses inflammation in murine model systems; for example, IL-10 is important in Hymenolepis diminuta-inhibition of DNBS-induced colitis. Bacteria-derived products can have anti-inflammatory effects. Given that infection with H. diminuta, or other parasitic worms, results in perturbation of the gut microbiota, the present study tested a role for bacteria in helminth-suppression of colitis by assessing reciprocity between IL-10 and butyrate signaling in the amelioration of colitis. Aims To determine if a functional relationship exists between IL-10 and butyrate in the inhibition of colitis observed following infection with the lumen-dwelling tapeworm, Hymenolepis diminuta. Methods Colitis was induced in male BALB/c mice by intra-rectal dinitrobenzene sulphonic acid (DNBS) (3 mg/~22g mouse), with necropsy and assessment 3 days later. Mice received either infection with five H. diminuta cysticercoids by gavage or daily butyrate enemas or acetate in their drinking water. Immunostaining assessed IL-10R protein expression on formalin-fixed sections of colon. The murine IEC4.1 epithelial cell line and epithelial organoids were treated with butyrate and mRNA for the IL10Rα chain assessed, as was colonic tissue from mice. Results Mice infected with H. diminuta or receiving butyrate enemas (n=8–12) were protected from DNBS-induced colitis as gauged by colon length, and macroscopic disease and histopathology scores. Addition of acetate to the drinking water resulted in a more modest anti-colitic effect. Suppression of colitis was accompanied by increased epithelial expression of IL-10 in butyrate- and H. diminuta-treated mice, with the later also showing upregulation of the IL-10R on lamina propria cells; an effect negated by co-treating the mice with broad spectrum antibiotics. In vitro analyses revealed increased IL10Rα mRNA in butyrate-treated epithelia (n=4). Conclusions This study begins to tease apart the host (i.e. IL-10) and bacterial (i.e. butyrate) molecules that mediate H. diminuta-evoked suppression of colitis in a murine model. These proof-of-principle data suggest that knowledge of the individual patient (i.e. immunological basis of their disease and their microbiota) may be a critical determinant of the success or failure of helminth therapy. Funding Agencies CAG, CCCNSERC

Binding of Aflatoxin B1 to Bifidobacteria In Vitro

2000 ◽  
Vol 63 (8) ◽  
pp. 1133-1136 ◽  
Author(s):  
JAIMIE T. OATLEY ◽  
MATTHEW D. RARICK ◽  
GEUN EOG JI ◽  
JOHN E. LINZ
Keyword(s):  
Aflatoxin B1 ◽  
Binding Capacity ◽  
Large Fraction ◽  
Strain Differences ◽  
Potential Method ◽  
Fermented Foods ◽  
Binding Assays ◽  
Gut Flora ◽  
Food And Feed

Aflatoxins are mycotoxins that cause health and economic problems when they contaminate food and feed. One potential method for reducing human health effects due to aflatoxin ingestion is to block uptake via binding by bacteria that either make up the normal gut flora or are present in fermented foods in our diet. These bacteria would bind aflatoxin and make it unavailable for absorption in the intestinal tract. Bifidobacteria comprise a large fraction of the normal gut flora, are thought to provide many probiotic effects and are increasingly used in fermented dairy products. These qualities targeted bifidobacteria for studies to determine if various strains of heat-killed bifidobacteria can bind aflatoxin B1 (AFB1) in vitro. The AFB1 binding affinities of various strains of bifidobacteria, Staphylococcus aureus, and Escherichia coli were quantitated utilizing enzyme-linked immunosorbent and [3H]AFB1 binding assays. The bacteria analyzed were found to bind significant quantities of AFB1 ranging from 25% to nearly 60% of the added toxin. The data also suggest that there are reproducible strain differences in AFB1 binding capacity.

scholarly journals Modulation of Porcine β-Defensins 1 and 2 upon Individual and Combined Fusarium Toxin Exposure in a Swine Jejunal Epithelial Cell Line

2013 ◽  
Vol 79 (7) ◽  
pp. 2225-2232 ◽  
Author(s):  
Murphy Lam-Yim Wan ◽  
Chit-Shing Jackson Woo ◽  
Kevin J. Allen ◽  
Paul C. Turner ◽  
Hani El-Nezami
Keyword(s):  
Epithelial Cell ◽  
Mrna Expression ◽  
Cell Line ◽  
Defense Mechanisms ◽  
Fumonisin B1 ◽  
Enzyme Linked Immunosorbent Assay ◽  
Epithelial Cell Line ◽  
Content Type ◽  
Antimicrobial Effects ◽  
Food And Feed

ABSTRACTDefensins are small antimicrobial peptides (AMPs) that play an important role in the innate immune system of mammals. Since the effect of mycotoxin contamination of food and feed on the secretion of intestinal AMPs is poorly understood, the aim of this study was to elucidate the individual and combined effects of four commonFusariumtoxins, deoxynivalenol (DON), nivalenol (NIV), zearalenone (ZEA), and fumonisin B1 (FB1), on the mRNA expression, protein secretion, and corresponding antimicrobial effects of porcine β-defensins 1 and 2 (pBD-1 and pBD-2) using a porcine jejunal epithelial cell line, IPEC-J2. In general, upregulation of pBD-1 and pBD-2 mRNA expression occurred following exposure toFusariumtoxins, individually and in mixtures (P< 0.05). However, no significant increase in secreted pBD-1 and pBD-2 protein levels was observed, as measured by enzyme-linked immunosorbent assay (ELISA). Supernatants from IPEC-J2 cells exposed to toxins, singly or in combination, however, possessed significantly less antimicrobial activity againstEscherichia colithan untreated supernatants. When single toxins and two-toxin combinations were assessed, toxicity effects were shown to be nonadditive (including synergism, potentiation, and antagonism), suggesting interactive toxin effects when cells are exposed to mycotoxin combinations. The results show thatFusariumtoxins, individually and in mixtures, activate distinct antimicrobial defense mechanisms possessing the potential to alter the intestinal microbiota through diminished antimicrobial effects. Moreover, by evaluating toxin mixtures, this improved understanding of toxin effects will enable more effective risk assessments for common mycotoxin combinations observed in contaminated food and feed.

scholarly journals Effect of Butyric Acid in the Proliferation and Migration of Junctional Epithelium in the Progression of Periodontitis: An In Vitro Study

2021 ◽  
Vol 9 (4) ◽  
pp. 44
Author(s):  
Taichi Ishikawa ◽  
Daisuke Sasaki ◽  
Ryo Aizawa ◽  
Yu Shimoyama ◽  
Matsuo Yamamoto ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Epithelial Cell ◽  
Butyric Acid ◽  
In Vitro Study ◽  
Epithelial Cell Line ◽  
Junctional Epithelium ◽  
Tissue Destruction ◽  
High Concentration ◽  
Term Enrichment

Purpose: To elucidate the effects of butyric acid (BA), a metabolite of bacteria involved in periodontitis, and a possible enhancer of the junctional epithelial cells. Methods: A murine junctional epithelial cell line, JE-1, was used to assess the effects of sodium butyrate (NaB) as BA. Cell proliferation, migration and attachment were analyzed. Additionally, gene and promoter expression analysis was performed, i.e., cap analysis of gene expression (CAGE) and gene ontology (GO) term enrichment analysis. Results: NaB affected junctional epithelial cell proliferation, migration and attachment. A high concentration of NaB caused cell death and a low concentration tended to promote migration and adhesion. CAGE analysis revealed 75 upregulated and 96 downregulated genes in the cells after 0.2 mM NaB stimulation for 3 h. Regarding GO term enrichment, the genes upregulated >4-fold participated predominantly in cell migration and proliferation. The results of this study suggest that BA produced from periodontopathic bacteria is involved in periodontal tissue destruction at high concentrations. Furthermore, at low concentrations, BA potentially participates in periodontal disease progression by increasing proliferation, migration and attachment of the junctional epithelium and thereby increasing epithelial down-growth.

scholarly journals Evaluation of safety and antifungal activity of Lactobacillus reuteri and Pediococcus diacetilactis isolates against aflatoxigenic Aspergillus flavus

2019 ◽  
Vol 22 (2) ◽  
pp. 190-199 ◽  
Author(s):  
A. Sadeghi ◽  
M. Ebrahimi ◽  
B. Sadeghi ◽  
S. A. Mortazavi
Keyword(s):  
Antifungal Activity ◽  
Aspergillus Flavus ◽  
Lactobacillus Reuteri ◽  
Clinical Chemistry ◽  
Antagonistic Effect ◽  
Pcr Products ◽  
Food And Feed ◽  
Chemical Preservatives

Biocontrol of moulds by lactic acid bacteria (LAB) instead of antibiotics and chemical preservatives is a new approach in veterinary medicine. The aims of present research were to perform molecular identification of dominant sourdough LAB isolates and to evaluate their in vivo safety and in vitro antifungal properties for using as biopreservative agents. Sequencing results of PCR products led to identification of Lactobacillus reuteri and Pediococcus diacetilactis as LAB isolates. Antifungal activity of the isolates and their cell-free culture filtrate (CCF) were also confirmed against aflatoxigenic Aspergillus flavus, respectively by overlay and spore spot methods. Accordingly, antagonistic effect of P. diacetilactis and its CCF were significantly (P<0.05) higher than L. reuteri and CCF of mentioned LAB isolate. Clinical chemistry and haematological findings in mice fed LAB demonstrated also insignificant difference vs control mice and were in the normal range, which confirmed the safety of LAB isolates. By considering the importance of safe, food grade biocontrol agents, L. reuteri and P. diacetilactis isolates and their CCF may be considered as an alternative for antibiotics and other chemical preservatives in food and feed processing chain

scholarly journals Response of Human Alveolar Epithelial (hAELVi) Cells to Heavy Metals in Airborne Particulate

2021 ◽  
Author(s):  
Hui-Fang Chang ◽  
Ji-Yen Cheng
Keyword(s):  
Heavy Metals ◽  
Heavy Metal ◽  
Metal Ion ◽  
Epithelial Cell Line ◽  
Ros Production ◽  
High Concentration ◽  
Permeable Membrane ◽  
Alveolar Epithelial ◽  
Human Lung Cells

Abstract In vitro human alveolar models provide a platform to investigate the cellular behavior and activity of human lung cells for toxicity assays. However, there has been relatively rare research on the new human alveolar epithelial cell line (hAELVi) for the toxicity induced by heavy metal ions. In this study, hAELVi cells were exposed to various concentrations of Cd, As, and Zn for 24h. The morphological changes of hAELVi cells were observed after exposure to heavy metals. A high concentration of Cd, As, and Zn led to cell death. However, the cytotoxicity was not reflected by the transepithelial electrical resistance (TEER) value after exposure to 250 μM of As and Zn. In addition, Cd at a concentration of 200 μM induced the significant production of TNF-α whereas As and Zn did not induce observable secretion of the cytokine. On the contrary, a high concentration of Cd, As, and Zn inhibited the production of IL-6. In addition, there is not a direct relationship between the metal ion cytotoxicity and the ROS production in the hAELVi cells. Accordingly, this study contributes to elucidating the cytotoxicity, TEER variation, ROS production, and cytokine secretion of hAELVi cells to acute exposure to heavy metal in vitro. The discrepancy between the cytotoxicity and the TEER result suggests that hAELVi cell cultured on permeable membrane exhibit very different behavior, which is worth pursuing.

scholarly journals Re-Evaluation of Combinational Efficacy and Synergy of the Italian Protocol In Vitro: Are We Truly Optimizing Benefit or Permitting Unwanted Toxicity?

Biomedicines ◽  
2021 ◽  
Vol 9 (9) ◽  
pp. 1190
Author(s):  
Chitra Subramanian ◽  
Reid McCallister ◽  
Dawn Kuszynski ◽  
Mark S. Cohen
Keyword(s):  
Synergistic Effect ◽  
Cell Lines ◽  
Synergistic Effects ◽  
Antagonistic Effect ◽  
Steroidogenic Enzymes ◽  
In Vivo Evaluation ◽  
Anticancer Efficacy ◽  
Lower Toxicity

Introduction: Adrenocortical carcinoma (ACC) is a rare endocrine malignancy, with very poor prognosis as a majority of the patients have advanced disease at the time of diagnosis. Currently, adjuvant therapy for most patients consists of either mitotane (M) alone or in combination with multi-drug chemotherapeutics such as etoposide (E), doxorubicin (D), and cisplatin (P), known as the Italian protocol (IP; EDPM). This multi-drug treatment regimen, however, carries significant toxicity potential for patients. One way to improve toxicity profiles with these drugs in combination is to understand where their synergy occurs and over what dosing range so that lower dose regimens could be applied in combination with equal or improved efficacy. We hypothesize that a better understanding of the synergistic effects as well as the regulation of steroidogenic enzymes during combination therapy may provide more optimized combinational options with good potency and lower toxicity profiles. Methods: Two human ACC cell lines, NCI-H295R (hormonally active) and SW13 (hormonally inactive), were grown in 2D culture in appropriate growth medium. The viability of the cells after treatment with varying concentrations of the drugs (E, D, and P) either alone or in combinations with M was determined using the CellTiter Glow assay after 72 h, and the combination index for each was calculated using Compusyn by the Chou–Talalay method. The expression levels of enzymes associated with steroidogenesis were evaluated by RT-PCR in NCI-H295R. Results: When both cell lines were treated with M (ranging 25–50 μM), +E (ranging 18.75–75 μM), and +D (ranging 0.625–2.5 μM) we observed a synergistic effect (CI < 1) with potency equivalent to the full Italian protocol (IP), whereas combining M + P + D had an antagonistic effect (CI > 1) indicating the negative effect of adding cisplatin in the combination. Comparing the hormonally active and inactive cell lines, M + P + E was antagonistic in NCI-H295R and synergistic in SW13. Treatment of NCI-H295R cells with antagonistic combinations (M + P + D, M + P + E) resulted in a significant decrease in the levels of steroidogenic enzymes STAR, CYP11A1, and CYP21A2 compared to IP (p < 0.05) while M + E + D resulted in increased expression or no significant effect compared to IP across all genes tested. Conclusions: The synergistic effect for M + E + D was significant and equivalent in potency to the full IP in both cell lines and resulted in a steroidogenic gene expression profile similar to or better than that of full IP, warranting further evaluation. Future in vivo evaluation of the combination of M + E + D (with removal of P from the IP regimen) may lower toxicity while maintaining anticancer efficacy in ACC.

scholarly journals Transcriptogram analysis reveals relationship between viral titer and gene sets responses during Corona-virus infection

2020 ◽  
Author(s):  
Rita M.C. de Almeida ◽  
Gilberto L. Thomas ◽  
James A. Glazier
Keyword(s):  
Time Course ◽  
Differentially Expressed Gene ◽  
Epithelial Cell Line ◽  
Mitochondrial Activity ◽  
Viral Titer ◽  
Individual Gene ◽  
Human Lung Epithelial Cell ◽  
Gene Sets ◽  
The Individual

AbstractTo understand the difference between benign and severe outcomes after Coronavirus infection, we urgently need ways to clarify and quantify the time course of tissue and immune responses. Here we re-analyze 72-hour time-series microarrays generated in 2013 by Sims and collaborators for SARS-CoV-1 in vitro infection of a human lung epithelial cell line. Using a Transcriptogram-based top-down approach, we identified three major, differentially-expressed gene sets comprising 219 mainly immune-response-related genes. We identified timescales for alterations in mitochondrial activity, signaling and transcription regulation of the innate and adaptive immune systems and their relationship to viral titer. At the individual-gene level, EGR3 was significantly upregulated in infected cells. Similar activation in T-cells and fibroblasts in infected lung could explain the T-cell anergy and eventual fibrosis seen in SARS-CoV-1 infection. The methods can be applied to RNA data sets for SARS-CoV-2 to investigate the origin of differential responses in different tissue types, or due to immune or preexisting conditions or to compare cell culture, organoid culture, animal models and human-derived samples.

Assays for Phospholipid-Dependent Formation of Thrombin and Xa: A Potential Method for Quantifying Lupus Anticoagulant Activity

1991 ◽  
Vol 66 (04) ◽  
pp. 453-458 ◽  
Author(s):  
John T Brandt
Keyword(s):  
Specific Activity ◽  
Anticoagulant Activity ◽  
Factor Xa ◽  
Clinical Importance ◽  
Potential Method ◽  
Quantitative Expression ◽  
Increased Risk ◽  
Apparent Association

SummaryLupus anticoagulants (LAs) are antibodies which interfere with phospholipid-dependent procoagulant reactions. Their clinical importance is due to their apparent association with an increased risk of thrombo-embolic disease. To date there have been few assays for quantifying the specific activity of these antibodies in vitro and this has hampered attempts to purify and characterize these antibodies. Methods for determining phospholipid-dependent generation of thrombin and factor Xa are described. Isolated IgG fractions from 7 of 9 patients with LAs were found to reproducibly inhibit enzyme generation in these assay systems, permitting quantitative expression of inhibitor activity. Different patterns of inhibitory activity, based on the relative inhibition of thrombin and factor Xa generation, were found, further substantiating the known heterogeneity of these antibodies. These systems may prove helpful in further purification and characterization of LAs.

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