Patulin Toxicity Research Articles

Cellulose-based magnetic nanomaterials immobilized esterases as a reusable and effective detoxification agent for patulin in apple juice

Patulin, an important contaminant in apple juice processing, exhibits various toxicities to humans. In existing patulin detoxification methods, enzymes are desirable biological detoxification agents. Here, magnetic nanocrystalline cellulose (MNCC) was prepared and employed as support for the immobilization of porcine liver esterase (PLE) to synthesize a novel cellulose-based magnetic patulin detoxification agent (PLE@MNCC). The detoxification rate of patulin by PLE@MNCC exceeded 80%. PLE@MNCC also enabled rapid separation and recovery after application while exhibiting superior storage stability, reusability and biocompatibility. After 8 repetitive cycles of usage, PLE@MNCC retained approximately 50% of its initial activity. Moreover, the cytotoxicity evaluation demonstrated a significant decrease in patulin toxicity after the detoxification. Crucially, there were no significant adverse changes in the quality of apple juice after detoxification. PLE@MNCC as an effective, reusable, environmentally friendly and safe enzyme agent provided an efficient strategy for the detoxification of patulin in fruit juices.

In Vitro and In Vivo Evaluation of Chlorogenic Acid-Encapsulated Lignin on Patulin Adsorption and Alleviation of Patulin-Induced Colonic Damage.

Patulin (PAT) is a water-soluble mycotoxin that causes digestive tract damage and liver and kidney function abnormalities. The current control approaches only reduce the amount of PAT in raw materials and food, which is difficult to remove once ingested in the body. In this study, lignin-based cross-linked particles loaded with chlorogenic acid were prepared, which intervened the liver and colon damage caused by PAT in mice. In the simulated digestion process in vitro, the accumulated adsorption capacity of the LB/CA-SH for PAT was 0.934 mg/g. LB/CA-SH intervention reversed the shortening of the colon length, alleviated the changes in the activities of antioxidant enzymes, and reduced the levels of oxidation markers protein carbonyl and malondialdehyde in the colon tissue of the model group. The absorption of sorbent alleviated the decrease of organ index and the abnormality of serum biochemical indexes (alanine aminotransferase, aspartate aminotransferase, urea nitrogen, and uric acid) caused by PAT. These results support the potential of using LB/CA-SH as a novel protective agent to reduce the toxicity of PAT.

Estimation of antigenotoxic properties of Ginger (Zingibe officinale) for DNA Damage after Exposure to Patulin in rats

Patulin, a known fungal poison, is an important pollutant in apples and products derived from apples and fodder. Ginger improved the liver function and prevents hepatotoxicity against many toxins and used in treatment of various diseases. The genetic toxicity of patulin in the liver of rats using the comet assay in vivo was investigated and effect of ginger (Zingibe officinale) on DNA damage induced by patulin. Five groups of adult male Sprague Dawley rats were used. The rats were given 3.75 mg kg-1 intraperitoneal (i.p) patulin. The rats were treated with ginger at a dose (100 mg kg-1) for four weeks and eight weeks. DNA damage was measured in the liver of rats using a standard genetic toxicity tests comet assay. Tail length and percentage of tail DNA within the comet increased significantly (p <0.05) in patulin group (p =0.01), (p =0.000) respectively, compared with control group a time-dependent manner, indicating an increase in DNA damage. The liver tissue showed signs of improvement in DNA damage after treating rats with ginger. In conclusion, these results indicate that patulin is genotoxic at the injected dose and that liver is an important target organ. Instead, ginger therapy can reduce DNA damage in liver cells from the toxicity caused by patulin.

The effects of natural active substances in food on the toxicity of patulin

Patulin (PAT) is a mycotoxin, a secondary metabolite mainly produced by fungi of the generaAspergillus, Byssochlamys, andPenicillium. Many studies have looked into the potential impacts of this mycotoxin due to its high risk. Researchers are currently doing a more in-depth investigation of and employing physical, chemical, and biological ways to remove PAT. However, existing technology cannot completely remove it, and the residual PAT will continue to pose a threat to human health. As a result, substances capable of reducing PAT toxicity need be discovered. According to previous studies, natural components in food could reduce the toxicity of PAT. This article will review the different types of active compounds and discus the detoxification processes, as well as give recommendations for decreasing the toxicity of PAT and future research directions.

Possible Reaction Mechanisms Involved in Degradation of Patulin by Heat-Assisted Cysteine under Highly Acidic Conditions

Patulin (PAT) is one of mycotoxins that usually contaminates apple juice, and it is not easily detoxified by cysteine (CYS) at room temperature due to the highly acidic conditions based on the Michael addition reaction. However, it could be effectively degraded by a heating treatment at 120 °C for 30 min in the presence of cysteine. In our study, a total of eight degradation products (DP A–H) were characterized and identified via liquid chromatography quadrupole time-of-flight mass spectrometry (LC-Q-TOF-MS) in a negative ion mode, and their structures and formulas were proposed based on their accurate mass data. The fragmentation patterns of PAT and its degradation products were obtained from the MS/MS analysis. Meanwhile, the possible reaction mechanisms involved in the degradation of PAT were established and explained for the first time. According to the relation between the structure and toxicity of PAT, it could be deduced that the toxic effects of PAT degradation products were potentially much less than those of PAT-self.

Toxicity mechanism of patulin on 293T cells and correlation analysis of Caspase family.

Patulin (PAT), a kind of mycotoxin, is a widely disseminated mycotoxin found in agricultural products. Although the existing research results show that PAT can cause nerve, immune, and skin toxicities, resulting in heart, liver, and kidney damages. However, evidence on the underlying mechanisms of PAT is still lacking. Present study aims to investigate the renal toxicity and related mechanisms of PAT on 293T cells. Cell Counting Kit-8 method was used to reveal the dose-effect relationship and the time-effect relationship of PAT toxicity. Trypan blue staining and Hoechst 33342 staining were used to analyze PAT, which induced apoptosis on 293T cells. Superoxide-dismutase (SOD), GSH, and malondialdehyde (MDA) were used to measure the changes of oxidative stress status of 293T cells induced by PAT. The changes of reactive oxygen species (ROS) and ATP in mitochondria indicate the role of mitochondria when PAT induced cell damage and apoptosis. Through Cyt-C release assay analysis, caspase activity change, and correlation analysis, the potential mechanism of mitochondrial apoptosis pathway was proved. Results demonstrated that PAT significantly induced cell injury, and with the increase of time and concentration, the cell survival rate decreased significantly. Hoechst 33342 staining and Trypan blue staining showed that apoptosis rate was elevated by PAT. As PAT concentration increased, intracellular SOD, glutathion peroxidase activities were decreased and the MDA content was increased. The decrease of intracellular ATP level and accumulation of ROS content indicated an increased permeability of the mitochondrial membrane. Overexpression of Cyt-C activated the cascade reaction of caspase enzyme, leading to apoptosis. The results of enzyme activity assay and correlation analysis indicated that caspase 3 was the most critical caspase in the cascade system and that it was most correlated with caspase 8 and caspase 9.

Selenium-Enriched Pediococcus acidilactici MRS-7 Alleviates Patulin-Induced Jejunum Injuries in Mice and Its Possible Mechanisms.

Patulin (PAT) is a common mycotoxin. Oral ingestion of PAT could damage the intestinal mucosa. Both selenium and probiotics can alleviate intestinal damage, but there are few reports on selenium-enriched probiotics. Here, we studied the protective effects of a new selenium-enriched Pediococcus acidilactici MRS-7 (SeP) on PAT-induced jejunum injuries in mice. Results show that PAT induced jejunum injuries such as loss of crypts, ulceration of the mucosa, and intestinal epithelial barrier function impairment. However, SeP could protect against PAT-induced jejunum injuries and significantly inhibit the reduction of goblet cell numbers. SeP could not only alleviate PAT-induced oxidative stress by decreasing malondialdehyde (MDA) and increasing superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px) levels in the jejunum tissues but also alleviate the inflammatory response caused by PAT by reducing the levels of inflammatory factors (interleukin (IL)-6 snd IL-1β and tumor necrosis factor-α (TNF-α)) in the serum and jejunum tissues. In addition, SeP also inhibited the expression of nuclear factor-κB (NF-κB) and Toll-like receptor 4 (TLR-4), increased the expression of tight junction proteins (occludin, ZO-1, and claudin-1), and increased the selenium content in the jejunum, thereby antagonizing the jejunum injuries caused by PAT exposure. Finally, SeP rebalanced the intestinal microbiota and improved probiotic abundance such as Turicibacter, Bifidobacterium, Ileibacterium, and Pediococcus in PAT-treated mice. These results support the possibility of SeP as a novel protective agent to mitigate the toxicity of PAT.

The Nrf1 transcription factor is induced by patulin and protects against patulin cytotoxicity

Patulin is a mycotoxin produced by a variety of molds that is found in various food products. The adverse health effects associated with exposure to patulin has led to many investigations into the biological basis driving the toxicity of patulin. Nevertheless, the mechanisms through which mammalian cells resists patulin-mediated toxicity is poorly understood. Here, we show that loss of the Nrf1 transcription factor renders cells sensitive to the acute cytotoxic effects of patulin. Nrf1 deficiency leads to accumulation of ubiquitinated proteins and protein aggregates in response to patulin exposure. Nrf1 expression is induced by patulin, and activation of proteasome genes by patulin is Nrf1-dependent. These findings suggest the Nrf1 transcription factor plays a crucial role in modulating cellular stress response against patulin cytotoxicity.

Effects of Selenium Nanoparticles on Preventing Patulin-Induced Liver, Kidney and Gastrointestinal Damage.

Patulin (PAT) is a toxic fungal metabolite, and oxidative damage was proved to be its important toxicity mechanism. Selenium nanoparticles (SeNPs) were prepared by reducing sodium selenite with chitosan as a stabilizer and used for preventing PAT-induced liver, kidney and gastrointestinal damage. SeNPs have good dispersibility, in vitro antioxidant activity, and are much less cytotoxic than sodium selenite. Cell culture studies indicated that SeNPs can effectively alleviate PAT-induced excessive production of intracellular ROS, the decline of glutathione peroxidase activity, and the suppression of cell viability. Evaluation of serum biochemical parameters, histopathology, oxidative stress biomarkers and activities of antioxidant enzymes in a mouse model showed that pre-treatment with SeNPs (2 mg Se/kg body weight) could ameliorate PAT-induced oxidative damage to the liver and kidneys of mice, but PAT-induced gastrointestinal oxidative damage and barrier dysfunction were not recovered by SeNPs, possibly because the toxin doses suffered by the gastrointestinal as the first exposed tissues exceeded the regulatory capacity of SeNPs. These results suggested that a combination of other strategies may be required to completely block PAT toxicity.

Toxic effects of patulin on mouse oocytes and its possible mechanisms

Patulin is a secondary metabolite mainly secreted by fungi and is the most common mycotoxin found in apples and apple-based products. For the past few years, numerous studies suggested the wide distribution and toxicity of patulin. In this study, we investigated the toxic effect of patulin on mouse oocytes and its possible mechanisms. The results showed that patulin treatment did not affect meiotic resumption, but inhibited oocyte maturation as indicated by failure of first polar body extrusion. Further mechanistic study showed that patulin treatment disturbed normal spindle assembly, chromosome alignment and morphology. We also found increased oxidative stress by testing the level of ROS and decreased mitochondrial membrane potential, indicating mitochondria dysfunction. In summary, our results suggest that patulin treatment causes oocyte meiotic arrest by disturbing normal spindle assembly and chromosome alignment, which may be caused by dysfunctions of mitochondria.

Natural Thiols, but Not Thioethers, Attenuate Patulin-Induced Endoplasmic Reticulum Stress in HepG2 Cells.

Patulin, a mycotoxin, is known to have cytotoxic effects, but few studies have focused on the involvement of the endoplasmic reticulum (ER) stress response in patulin toxicity and the natural compounds that attenuate it in HepG2 cells. This study tested the ability of patulin to induce ER stress, and that of four thiols and three thioethers to attenuate patulin-induced ER stress in HepG2 cells. Patulin dose-dependently inhibited cell proliferation (IC50, 8.43 μM). Additionally, patulin was found to increase the expression levels of ER stress-related genes and/or protein markers, including BiP, CHOP, and spliced XBP1, in HepG2 cells compared to the vehicle control, indicating its potential in ER stress induction. Patulin-induced cytotoxicity in HepG2 cells was reduced by naturally occurring thiol compounds (glutathione, L-acetyl-L-cysteine, cysteine, and captopril), but not by thioether compounds (sulforaphane, sulforaphene, and S-allyl-L-cysteine). Patulin-thiol co-treatment decreased CHOP expression and BiP and CHOP levels in HepG2 cells but did not alter BiP expression. Spliced XBP1 expression was decreased by patulin-thiol co-treatment. Thus, patulin induced ER stress in HepG2 cells and thiols, but not in thioethers, attenuated patulin-induced ER stress.

Effect of Rhodotorula mucilaginosa on patulin degradation and toxicity of degradation products

ABSTRACT Rhodotorula mucilaginosa is an antagonistic yeast for which our research team has recently reported interesting biocontrol activities against blue mould decay of apples and a strong ability to decrease the patulin concentration in vivo. However, the possible mechanisms of patulin degradation by R. mucilaginosa and the toxicity of patulin degradation products remain unclear. In this study, the effect of R. mucilaginosa on patulin degradation and toxicity of degradation products were investigated, the results showed that viable cells of R. mucilaginosa are essential to patulin degradation. Also, R. mucilaginosa eliminated patulin without adsorbing it through its cell wall. The extracellular metabolites of R. mucilaginosa stimulated by patulin showed little degradation activity for patulin. Cycloheximide addition into the medium significantly decreased the patulin degradation capacity of R. mucilaginosa cells. The main patulin degradation product by R. mucilaginosa was ascladiol, which was proved non-toxic to human hepatoma (HepG2) cells at 0.625–10 g/mL. Furthermore, toxicological analysis using a confocal laser scanning microscope revealed that the degradation product induced cellular apoptosis to a lesser extent than patulin itself. This result offers an innovative method to detoxify patulin and limit the risks of patulin in fruits and vegetables using R. mucilaginosa.

Insights into the highly efficient detoxification of the biotoxin patulin in water by discharge plasma oxidation

Patulin is a neurotoxic secondary metabolite and is widely present in fruit-based products and even in drinking water. The low-temperature discharge plasma approach was developed to detoxify patulin in water. Patulin decomposition and mineralization efficiencies reached 93% and 74% at an applied voltage of 16 kV in a 30 min treatment period, respectively. 1O2, OH, O2−, and hydrated electrons generated in the discharge plasma process contributed to patulin decomposition. The lactone and hemiacetal groups that are responsible for the toxicity of patulin were significantly destroyed by discharge plasma oxidation, and some ring-opening intermediates, such as ethers, organic acids, and alcohols, were generated. Patulin decomposition pathways in this low-temperature discharge plasma process were then proposed. Model prediction indicated that acute oral toxicity in rats and developmental toxicity decreased after discharge plasma treatment, which was verified by actual cytotoxicity tests using yeast cells and HepG2 human hepatocellular carcinoma cells.

Transcriptomic and Proteomic Analysis Reveals Mechanisms of Patulin-Induced Cell Toxicity in Human Embryonic Kidney Cells.

Patulin (PAT) is a natural mycotoxin that commonly contaminates fruits and fruit-based products. Previous work indicated that PAT-induced apoptosis in which reactive oxygen species (ROS) are involved in human embryonic kidney (HEK293) cells. To uncover novel aspects of the possible mechanism of PAT nephrotoxicity, the transcriptome and proteome profiles were investigated using the digital gene expression (DGE) and isobaric tags for relative and absolute quantitation (iTRAQ) proteomic approaches. A total of 127 genes and 85 proteins were found to express differentially in response to 5 μM PAT for 10 h in HEK293 cells. The most dramatic changes of expression were noticed with genes or proteins related to apoptosis, oxidative phosphorylation ribosome and cell cycle. Especially, the activation of caspase 3, UQCR11, active transport form and endocytosis appeared to be crucial in PAT kidney cytotoxicity. PAT also seemed to be associated with cancer and neuropathic disease as pathways associated with carcinogenesis, Alzheimer’s disease and Parkinson’s disease were induced. Overall, this study served to uncover overall insights associated with signaling pathway that modulated the PAT toxicity mechanism.

The characteristics of patulin detoxification by Lactobacillus plantarum 13M5

Patulin (PAT) is a widespread mycotoxin that harms the health of both humans and animals. In this study, among the 17 tested Lactobacillus plantarum strains, L. plantarum 13M5, isolated from traditional Chinese fermented foods, showed the highest PAT degradation rate of up to 43.8% (PAT 5 mg/L). Evaluation of the living and dead 13M5 cells revealed that only the living cells had the ability to remove PAT and degrade it into E-ascladiol. A cell-based assay revealed that L. plantarum 13M5 administration alleviated PAT-induced injuries in Caco-2 cells, including cytotoxicity, oxidative stress, and tight junction disruption. Our results suggest that L. plantarum 13M5 has the potential to reduce PAT toxicity and can thus be used as a probiotic supplement to reduce or eliminate the toxicity of PAT ingested from diet.

Targeting mitochondrial metabolite transporters in Penicillium expansum for reducing patulin production

There is an increasing need of alternative treatments to control fungal infection and consequent mycotoxin accumulation in harvested fruits and vegetables. Indeed, only few biological targets of antifungal agents have been characterized and can be used for limiting fungal spread from decayed fruits/vegetables to surrounding healthy ones during storage. On this concern, a promising target of new antifungal treatments may be represented by mitochondrial proteins due to some species-specific functions played by mitochondria in fungal morphogenesis, drug resistance and virulence.One of the most studied mycotoxins is patulin produced by several species of Penicillium and Aspergillus genera. Patulin is toxic to many biological systems including bacteria, higher plants and animalia. Although precise biochemical mechanisms of patulin toxicity in humans are not completely clarified, its high presence in fresh and processed apple fruits and other apple-based products makes necessary developing a strategy for limiting its presence/accumulation. Patulin biosynthetic pathway consists of an enzymatic cascade, whose first step is represented by the synthesis of 6-methylsalicylic acid, obtained from the condensation of one acetyl-CoA molecule with three malonyl-CoA molecules. The most abundant acetyl-CoA precursor is represented by citrate produced by mitochondria. In the present investigation we report about the possibility to control patulin production through the inhibition of mitochondrial/peroxisome transporters involved in the export of acetyl-CoA precursors from mitochondria and/or peroxisomes, with specific reference to the predicted P. expansum mitochondrial Ctp1p, DTC, Sfc1p, Oac1p and peroxisomal PXN carriers.

Hepato-renal Toxicity of Patulin and its Modulation by Ginger (Zingiber officinale) in Rats

Hepato-renal Toxicity of Patulin and its Modulation by Ginger (Zingiber officinale) in Rats

Distinct Transcriptional Changes in Response to Patulin Underlie Toxin Biosorption Differences in Saccharomyces Cerevisiae.

Patulin (4-hydroxy-4H-furo[3,2c]pyran-2[6H]-one) is a mycotoxin produced by a suite of fungi species. Patulin is toxic to humans and is a sporadic contaminant in products that were made from fungi-infected fruits. The baker yeast Saccharomyces cerevisiae (S. cerevisiae) has been shown to decrease patulin levels likely by converting it to the less harmful E-ascladiol, yet this capacity is dependent on the strain utilized. In this study we show that four representative strains of different S. cerevisiae lineages differ in their ability to tolerate and decrease patulin levels in solution, demonstrating that some strains are better suitable for patulin biocontrol. Indeed, we tested the biocontrol capacities of the best patulin-reducer strain (WE) in contaminated apple juice and demonstrated their potential role as an efficient natural biocontrol solution. To investigate the mechanisms behind the differences between strains, we explored transcriptomic changes of the top (WE strain) and worst (WA strain) patulin-biocontroller strains after being exposed to this toxin. Large and significant gene expression differences were found between these two strains, the majority of which represented genes associated with protein biosynthesis, cell wall composition and redox homeostasis. Interestingly, the WE isolate exhibited an overrepresentation of up-regulated genes involved in membrane components, suggesting an active role of the membrane towards patulin detoxification. In contrast, WA upregulated genes were associated with RNA metabolism and ribosome biogenesis, suggesting a patulin impact upon transcription and translation activity. These results suggest that different genotypes of S. cerevisiae encounter different stresses from patulin toxicity and that different rates of detoxification of this toxin might be related with the plasma membrane composition. Altogether, our data demonstrates the different molecular mechanisms in S. cerevisiae strains withstanding patulin exposure and opens new avenues for the selection of new patulin biocontroller strains.

Occurrence of patulin and 5-hydroxymethylfurfural in apple sour, which is a traditional product of Kastamonu, Turkey

ABSTRACTApple sour is a traditional product of Kastamonu, Turkey. It is consumed by spreading on bread or drinking after diluting with water. The aim of this study was to determine patulin (PAT) and 5-hydroxymethylfurfural (HMF) in apple sour. This study is the first to evaluate the occurrence of PAT and HMF in apple sour. The samples were extracted with ethyl acetate using liquid-liquid extraction technique. PAT and HMF were determined by HPLC with UV detection. PAT was detected in all samples, and the PAT level in 94.9% of samples was found to be equal or greater than the legal limit for juice concentrates. The mean value for PAT was found to be 284 ± 307 μg kg−1. PAT levels in 13 of 39 samples were in the range of 100 ≤ x < 200 μg kg−1, two samples were in the range of 0 ≤ x < 50 μg kg −1 and two samples were in the range of 1000 ≤ x < 1500 μg kg1. HMF levels of all samples were above the legal limit for solid molasses. The mean value for HMF was found to be 16215 ± 13317 mg kg−1. HMF levels of 10 of 39 samples were determined to be in the range of 10000 ≤ x < 20000 mg kg−1, eight samples were in the range of 20000 ≤ x < 30000 mg kg−1 and only three samples were in the range of 100 ≤ x < 1000 mg kg−1. There was a significant and inverse relationship between HMF and pH of the samples. These results indicate that consumption of apple sour is a considerable risk in terms of HMF and PAT toxicity.

Food-borne patulin toxicity is related to gut barrier disruption and can be prevented by docosahexaenoic acid and probiotic supplementation.

Patulin (PAT) is a mycotoxin widely found in fruits and vegetables. Several reviews and studies have hypothesized that in vivo PAT toxicity is related to gut barrier dysfunction, but evidence for this is not substantial. The goal of the present study was to further demonstrate the role of the gut barrier in food-borne PAT toxicity. In vitro assays showed that PAT exposure induced significant cell death, inhibited the mRNA expressions of tight junction proteins and increased gut permeability in Caco-2 cell monolayers. An acute PAT exposure animal trial reported for the first time an association between PAT-induced disruption of the gut barrier and endotoxemia in mice. Sub-chronic PAT exposure also inhibited the expression of ZO-1 in the gut and induced both intestinal and systematic inflammation in mice. Dietary supplements with previously reported protective effects on the gut barrier, such as docosahexaenoic acid and Lactobacillus plantarum CCFM8610, were able to recover the PAT-induced gut barrier dysfunction and significantly alleviate PAT toxicity in vivo. Another L. plantarum strain, CCFM11, with poor gut barrier modulation ability, failed to exhibit identical protective effects against PAT toxicity to L. plantarum CCFM8610. Our results indicated that PAT-induced disruption of the gut barrier and bacterial translocation may be another toxic mechanism of PAT besides its inherent cytotoxicity. Gut barrier protection may be considered an important target for the prevention of PAT toxicity.