scholarly journals Bioaccessibility of ochratoxin A from red wine in an in vitro dynamic gastrointestinal model

2015 ◽  
Vol 8 (1) ◽  
pp. 107-112 ◽  
Author(s):  
C.A. González-Arias ◽  
I. Piquer-Garcia ◽  
S. Marin ◽  
V. Sanchis ◽  
A.J. Ramos
Keyword(s):  
Ochratoxin A ◽  
Red Wine ◽  
Dialysis Membrane ◽  
Fluorescence Detector ◽  
Penicillium Species ◽  
In Vitro System ◽  
Small Doses ◽  
Flow Dialysis

Ochratoxin A (OTA) is a mycotoxin produced by Aspergillus and Penicillium species with immunosuppressive, teratogenic, and carcinogenic properties. It has been determined that wine is the second largest source of OTA (10% of total OTA intake) in the European diet and that its presence, even in small doses, can be a problem in terms of long-term toxicity. In this paper, we evaluated the bioaccessibility of OTA in a spiked red wine sample under human fasting conditions using an in vitro dynamic digestion model that includes a continuous-flow dialysis system to simulate intestinal passage. To the best of our knowledge, this report is the first examining the bioaccessibility of OTA in wine. A liquid-liquid method was used to extract the OTA and ochratoxin alpha (OTα) from gastrointestinal juices, and the extracts were analysed by HPLC with a fluorescence detector. The bioaccessibility of OTA from the spiked red wine (1.0, 2.0 and 4 μg/l) was high in the gastric compartment (102.8, 128.3 and 122.3%, respectively), whereas in the simulated intestine, it did not exceed 26%, and the amount of OTA that crossed the dialysis membrane was very low (<3.3%). The amount of OTα in gastric chyme ranged from 5.1 to 19.1% of the spiked OTA, whereas in the intestinal compartment it did not exceed 5%. In conclusion, in the in vitro system assayed, OTA exhibited a high bioaccessibility in the simulated stomach, but it decreased after the intestinal digestion and passage through the dialysis membrane.

Long-Term Dynamic in vitro System for Investigating Rat Pineal Melatonin Secretion

1991 ◽  
Vol 3 (5) ◽  
pp. 563-568 ◽  
Author(s):  
Z. Rékési ◽  
V. Csernus ◽  
J. Horvéth ◽  
S. Vigh ◽  
B. Mess
Keyword(s):  
Melatonin Secretion ◽  
In Vitro System ◽  
Pineal Melatonin

scholarly journals Human Proximal Tubule Epithelial Cells (HK-2) as a Sensitive In Vitro System for Ochratoxin A Induced Oxidative Stress

Toxins ◽  
2021 ◽  
Vol 13 (11) ◽  
pp. 787
Author(s):  
Enrique García-Pérez ◽  
Dojin Ryu ◽  
Hwa-Young Kim ◽  
Hae Dun Kim ◽  
Hyun Jung Lee
Keyword(s):  
Oxidative Stress ◽  
Ochratoxin A ◽  
Cell Lines ◽  
Time Dependent ◽  
Mrna Levels ◽  
Dependent Manner ◽  
In Vitro System ◽  
Glutathione Peroxidase 1 ◽  
Glucose 6 Phosphate Dehydrogenase

Ochratoxin A (OTA) is a mycotoxin that is potentially carcinogenic to humans. Although its mechanism remains unclear, oxidative stress has been recognized as a plausible cause for the potent renal carcinogenicity observed in experimental animals. The effect of OTA on oxidative stress parameters in two cell lines of LLC-PK1 and HK-2 derived from the kidneys of pig and human, respectively, were investigated and compared. We found that the cytotoxicity of OTA on LLC-PK1 and HK-2 cells was dose- and time-dependent in both cell lines. Furthermore, increased intracellular reactive oxygen species (ROS) induced by OTA in both cell lines were observed in a time-dependent manner. Glutathione (GSH) was depleted by OTA at >48 h in HK-2 but not in LLC-PK1 cells. While the mRNA levels of glucose-6-phosphate dehydrogenase (G6PD) and glutathione peroxidase 1 (GPX1) in LLC-PK1 were down-regulated by 0.67- and 0.66-fold, respectively, those of catalase (CAT), glutathione reductase (GSR), and superoxide dismutase 1 (SOD) in HK-2 were up-regulated by 2.20-, 2.24-, and 2.75-fold, respectively, after 72 h exposure to OTA. Based on these results, we conclude that HK-2 cells are more sensitive to OTA-mediated toxicity than LLC-PK1, and OTA can cause a significant oxidative stress in HK-2 as indicated by changes in the parameter evaluated.

A functional long‐term 2D serum‐free human hepatic in vitro system for drug evaluation

2020 ◽  
Author(s):  
Carlota Oleaga ◽  
L. Richard Bridges ◽  
Keisha Persaud ◽  
Christopher W. McAleer ◽  
Christopher J. Long ◽  
...  
Keyword(s):  
Drug Evaluation ◽  
In Vitro System ◽  
Serum Free

Patulin production by Penicillium expansum isolates from apples during different steps of long-term storage

2016 ◽  
Vol 9 (3) ◽  
pp. 379-388 ◽  
Author(s):  
N. De Clercq ◽  
G. Vlaemynck ◽  
E. Van Pamel ◽  
D. Colman ◽  
M. Heyndrickx ◽  
...  
Keyword(s):  
Phenotypic Diversity ◽  
Penicillium Expansum ◽  
Penicillium Species ◽  
Cold Temperatures ◽  
Long Term Storage ◽  
In Vitro Investigation ◽  
Duration Of Storage ◽  
Term Storage

Penicillium expansum is the principal cause of blue mould rot and associated production of patulin, a weak mycotoxin, in apples worldwide. P. expansum growth and patulin production is observed during improper or long-term storage of apples. We have investigated the extent to which each successive step during long-term storage contributes to patulin production in various P. expansum isolates. Fungal isolates collected on apples from several Belgian orchards/industries were identified to species level. Random amplification of polymorphic DNA (RAPD) analysis and β-tubulin gene sequencing identified P. expansum and Penicillium solitum as the most prevalent Penicillium species associated with Belgian apples. All 27 P. expansum isolates and eight reference strains were characterised for their patulin production capacity on apple puree agar medium for five days under classical constant temperature and atmosphere conditions. Under these conditions, a large range of patulin production levels was observed. Based on this phenotypic diversity, five P. expansum isolates and one reference strain were selected for in vitro investigation of patulin production under representative conditions in each step of long-term apple storage. Patulin accumulation seemed highly strain dependent and no significant differences between the storage steps were observed. The results also indicated that a high spore inoculum may lead to a strong patulin accumulation even at cold temperatures (1 °C) combined with controlled atmosphere (CA) (3% O2, 1% CO2), suggesting that future control strategies may benefit from considering the duration of storage under CA conditions as well as duration of deck storage.

scholarly journals Does Maternal Stress Affect the Early Embryonic Microenvironment? Impact of Long-Term Cortisol Stimulation on the Oviduct Epithelium

2020 ◽  
Vol 21 (2) ◽  
pp. 443 ◽  
Author(s):  
Shuaizhi Du ◽  
Nares Trakooljul ◽  
Jennifer Schoen ◽  
Shuai Chen
Keyword(s):  
Maternal Stress ◽  
Stress Hormones ◽  
Perceived Threat ◽  
In Vitro System ◽  
Expression Of Genes ◽  
Oviduct Epithelium ◽  
Air Liquid Interface ◽  
Oviductal Fluid

Maternal stress before or during the sensitive preimplantation phase is associated with reproduction failure. Upon real or perceived threat, glucocorticoids (classic stress hormones) as cortisol are synthesized. The earliest “microenvironment” of the embryo consists of the oviduct epithelium and the oviductal fluid generated via the epithelial barrier. However, to date, the direct effects of cortisol on the oviduct are largely unknown. In the present study, we used a compartmentalized in vitro system to test the hypothesis that a prolonged stimulation with cortisol modifies the physiology of the oviduct epithelium. Porcine oviduct epithelial cells were differentiated at the air–liquid interface and basolaterally stimulated with physiological levels of cortisol representing moderate and severe stress for 21 days. Epithelium structure, transepithelial bioelectric properties, and gene expression were assessed. Furthermore, the distribution and metabolism of cortisol was examined. The polarized oviduct epithelium converted basolateral cortisol to cortisone and thereby reduced the amount of bioactive cortisol reaching the apical compartment. However, extended cortisol stimulation affected its barrier function and the expression of genes involved in hormone signaling and immune response. We conclude that continuing maternal stress with long-term elevated cortisol levels may alter the early embryonic environment by modification of basic oviductal functions.

Interference of carbamylated and acetylated hemoglobins in assays of glycohemoglobin by HPLC, electrophoresis, affinity chromatography, and enzyme immunoassay

1993 ◽  
Vol 39 (1) ◽  
pp. 138-142 ◽  
Author(s):  
C W Weykamp ◽  
T J Penders ◽  
C W Siebelder ◽  
F A Muskiet ◽  
W van der Slik
Keyword(s):  
Affinity Chromatography ◽  
Enzyme Immunoassay ◽  
Serum Urea ◽  
Acetyl Coa ◽  
Total Hemoglobin ◽  
Small Doses ◽  
Significant Interference

Abstract In vitro-synthesized carbamylated and acetylated hemoglobins interfered in assays of glycohemoglobin by HPLC and electrophoresis but had no effects on results obtained by affinity chromatography and enzyme immunoassay. Correlations between long-term serum urea concentrations and glycohemoglobin percentages revealed that, in vivo, carbamylated hemoglobin equivalent to 0.063% of total hemoglobin is formed for every 1 mmol/L of serum urea. The use of acetylsalicylate, either chronically in small doses (200-300 mg/day) or for 1 week at 2000 mg/day, did not cause significant interference from acetylhemoglobin, formed in vivo. We conclude that interference from carbamylated hemoglobin explains only a small part of existing discrepancies between results of glycohemoglobin assays in current use. The interfering effect of acetylhemoglobin formed in vivo with acetyl-CoA as substrate is as yet unknown.

scholarly journals Exploiting oxidative phosphorylation to promote the stem and immunoevasive properties of pancreatic cancer stem cells

2020 ◽  
Vol 11 (1) ◽  
Author(s):  
Sandra Valle ◽  
Sonia Alcalá ◽  
Laura Martin-Hijano ◽  
Pablo Cabezas-Sáinz ◽  
Diego Navarro ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cancer Stem Cells ◽  
Ductal Adenocarcinoma ◽  
In Vitro System ◽  
Tumorigenic Potential ◽  
Term Enrichment ◽  
Pluripotency Gene ◽  
Pancreatic Cancer Stem Cells

Abstract Pancreatic ductal adenocarcinoma (PDAC), the fourth leading cause of cancer death, has a 5-year survival rate of approximately 7–9%. The ineffectiveness of anti-PDAC therapies is believed to be due to the existence of a subpopulation of tumor cells known as cancer stem cells (CSCs), which are functionally plastic, and have exclusive tumorigenic, chemoresistant and metastatic capacities. Herein, we describe a 2D in vitro system for long-term enrichment of pancreatic CSCs that is amenable to biological and CSC-specific studies. By changing the carbon source from glucose to galactose in vitro, we force PDAC cells to utilize OXPHOS, resulting in enrichment of CSCs defined by increased CSC biomarker and pluripotency gene expression, greater tumorigenic potential, induced but reversible quiescence, increased OXPHOS activity, enhanced invasiveness, and upregulated immune evasion properties. This CSC enrichment method can facilitate the discovery of new CSC-specific hallmarks for future development into targets for PDAC-based therapies.

scholarly journals Amplification of Sca-1+ Lin-WGA+ cells in serum-free cultures containing steel factor, interleukin-6, and erythropoietin with maintenance of cells with long-term in vivo reconstituting potential

Blood ◽  
1994 ◽  
Vol 83 (1) ◽  
pp. 128-136 ◽  
Author(s):  
VI Rebel ◽  
W Dragowska ◽  
CJ Eaves ◽  
RK Humphries ◽  
PM Lansdorp
Keyword(s):  
Interleukin 6 ◽  
Cell Number ◽  
In Vitro System ◽  
Serum Free ◽  
Murine Bone Marrow ◽  
Steel Factor ◽  
Parallel Increase

Abstract Normal murine bone marrow (BM) cells were sorted on the basis of low forward and orthogonal light scatter properties, Sca-1 expression (Sca-1+), lack of staining with a cocktail of mature hematopoietic lineage markers (Lin-), and binding of wheat germ agglutinin (WGA+). This approach allowed the reproducible isolation of a very small subpopulation (0.037% +/-0.023% of all nucleated BM cells) that was approximately 400-fold enriched in cells capable of reconstituting both lymphoid and myeloid lineages in lethally irradiated recipients. Transplantation of 30 or 10 of these Sca-1+Lin-WGA+ cells resulted in > or = to 20% donor-derived nucleated peripheral blood cells 3 months posttransplantation in 100% and 22% of the recipients, respectively. When Sca-1+Lin-WGA+ cells were cultured in serum-free medium supplemented with Steel factor, interleukin-6 (IL-6), and erythropoietin (with or without IL-3), a large increase in total cell number, including cells with an Sca-1+Lin-WGA+ phenotype was observed. Single cell cultures showed that 90% to 95% of the input cells underwent at least one division during the first 2 weeks and the remainder died. Interestingly, this proliferative response was not accompanied by a parallel increase in the number of cells with both lymphoid and myeloid repopulating potential in vivo, as quantitation of these by limiting dilution analysis showed they had decreased slightly (1.3-fold) but not significantly below the number initially present. These results demonstrate that Sca-1+Lin-WGA+ cells with long-term repopulating potential can be maintained for 2 weeks in a serum-and stroma cell-free culture, providing a simple in vitro system to study their behavior under well-defined conditions. The observed expansion of Sca-1+Lin-WGA+ cells in vitro without a concomitant increase in reconstituting cells also shows that extensive functional heterogeneity exists within populations of cells with this surface phenotype.

The FRTL-5 thyroid cell strain as a model for studies on thyroid cell growth

1987 ◽  
Vol 116 (1_Suppl) ◽  
pp. S242-S245 ◽  
Author(s):  
Francesco Saverio Ambesi-Impiombato ◽  
Giovanni Villone
Keyword(s):  
Cell Proliferation ◽  
Cell Growth ◽  
Cell Strain ◽  
Thyroid Cell ◽  
In Vitro System ◽  
Defined Media ◽  
Thyroid Follicular Cells ◽  
Rat Thyroid

Abstract. Thyroid cell proliferation has been studied using an in vitro system of rat thyroid follicular cell strain (FRTL-5). While growing in continuous culture, this strain is still differentiated and non-tumourigenic. Both advantages and limitations in the use of such system for studies of thyroid cell growth should be considered. Some obvious limitations should be considered, such as the species (rat) from which FRTL-5 cells were originated, their long-term growth outside the animals, the presence of a chronic TSH stimulation. On the other hand, several advantages as the growth in hormonally and chemically defined media, their dependence upon TSH in the medium, their genetic homogeneity and their widespread use in many laboratories render the FRTL-5 strain a useful experimental tool. Studies on cell proliferation and mechanism of action of hormones, growth factors and human autoimmune IgG have been and are being performed, with the assumption that FRTL-5 cells are the in vitro equivalent of thyroid follicular cells.

scholarly journals Amplification of Sca-1+ Lin-WGA+ cells in serum-free cultures containing steel factor, interleukin-6, and erythropoietin with maintenance of cells with long-term in vivo reconstituting potential

Blood ◽  
1994 ◽  
Vol 83 (1) ◽  
pp. 128-136 ◽  
Author(s):  
VI Rebel ◽  
W Dragowska ◽  
CJ Eaves ◽  
RK Humphries ◽  
PM Lansdorp
Keyword(s):  
Interleukin 6 ◽  
Cell Number ◽  
In Vitro System ◽  
Serum Free ◽  
Murine Bone Marrow ◽  
Steel Factor ◽  
Parallel Increase

Normal murine bone marrow (BM) cells were sorted on the basis of low forward and orthogonal light scatter properties, Sca-1 expression (Sca-1+), lack of staining with a cocktail of mature hematopoietic lineage markers (Lin-), and binding of wheat germ agglutinin (WGA+). This approach allowed the reproducible isolation of a very small subpopulation (0.037% +/-0.023% of all nucleated BM cells) that was approximately 400-fold enriched in cells capable of reconstituting both lymphoid and myeloid lineages in lethally irradiated recipients. Transplantation of 30 or 10 of these Sca-1+Lin-WGA+ cells resulted in > or = to 20% donor-derived nucleated peripheral blood cells 3 months posttransplantation in 100% and 22% of the recipients, respectively. When Sca-1+Lin-WGA+ cells were cultured in serum-free medium supplemented with Steel factor, interleukin-6 (IL-6), and erythropoietin (with or without IL-3), a large increase in total cell number, including cells with an Sca-1+Lin-WGA+ phenotype was observed. Single cell cultures showed that 90% to 95% of the input cells underwent at least one division during the first 2 weeks and the remainder died. Interestingly, this proliferative response was not accompanied by a parallel increase in the number of cells with both lymphoid and myeloid repopulating potential in vivo, as quantitation of these by limiting dilution analysis showed they had decreased slightly (1.3-fold) but not significantly below the number initially present. These results demonstrate that Sca-1+Lin-WGA+ cells with long-term repopulating potential can be maintained for 2 weeks in a serum-and stroma cell-free culture, providing a simple in vitro system to study their behavior under well-defined conditions. The observed expansion of Sca-1+Lin-WGA+ cells in vitro without a concomitant increase in reconstituting cells also shows that extensive functional heterogeneity exists within populations of cells with this surface phenotype.

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