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Abstract Patulin (PAT), a mycotoxin, is a natural contaminant that is produced by certain species of Penicillium, Aspergillus and Byssochlamys. The major contamination of PAT is in apple and apple based products. PAT is known to cause glutathione depletion, oxidative DNA damage and cell proliferation. Recently, in vitro studies have indicated that PAT can also increase the intestinal epithelial permeability, modulate tight junctions and decrease transepithelial electrical resistance. Nonetheless, no previous study has evaluated the mechanisms responsible for PAT-induced intestinal toxicity or its relevance to the in vivo situation. Here, Wistar rats were orally treated with 100 μg/kg body weight (b.wt.) of PAT, either alone or along with 100 mg/kg b. wt. of celecoxib for 3 days. We found that PAT exposure led to significantly higher levels of PGE 2 in serum and intestinal tissue and high expression of COX-2 and Ki-67 compared to controls. Interestingly, our results showed that celecoxib treatment could decrease the PAT-induced PGE 2 and reduce the PAT-induced intestinal damage. To study the mechanistic aspect, normal rat intestinal epithelial cells (IEC-6) were treated with non-toxic concentrations (100 nM, 250 nM and 500 nM) of PAT for 6 h. It was observed that PAT exposure caused enhanced proliferation, higher expression of COX-2, and EP2 and EP4 receptors, along with increased PGE 2 secretion. Additionally, PAT exposure caused enhanced Akt expression, which in turn inhibits GSK-3β and stabilizes β-catenin. Overall, our study suggests that the COX-2/EP2-EP4/β-catenin signaling cascades are involved in the regulation of PAT-induced intestinal cell proliferation and inflammation. Graphical abstract Unlabelled Image Highlights • Patulin exposure causes intestinal cell proliferation and inflammation. • Patulin causes over-expression of COX-2 and PGE 2 secretion. • EP2 and EP4 receptors are involved in the induction of COX-2 and PGE 2 secretion. • PGE 2 /EP2-EP4/β-catenin signaling regulates patulin induced intestinal toxicity. [ABSTRACT FROM AUTHOR]

Patulin (PAT) is a secondary metabolite produced by certain species of Penicillium, Byssochlamys and Aspergillus. It has been shown to induce liver toxicity, but the possible molecular mechanisms are not completely elucidated. In our study, we treated Human Hepatoma G2 (HepG2) cells by 3-methyladenine (3-MA), an autophagosome formation inhibitor, and rapamycin, an autophagosome formation stimulator. The results showed that 3-MA protected the HepG2 cells against PAT cytotoxicity, while rapamycin decreased the cell viability. Thus, autophagy may play an important role in PAT-induced toxicity. To uncover the mechanism by which cells decrease proliferation and activation of autophagy, we found that collapses of mitochondrial membrane potential (ΔΨm) and reactive oxygen species (ROS) level were increased under treatment with PAT. Further, we elucidated that the expression of p-Akt1 and p-MTOR was inhibited during this process. N-acetyl- l -cysteine (NAC), a ROS inhibitor, protected against PAT-induced cytotoxicity, decreased the protein expression of LC3-II, and up-regulated the level of p-Akt1 and p-MTOR. These findings suggested that PAT-induced autophagic cell death was ROS-dependent in HepG2 cells. In conclusion, it is possible that PAT elicited autophagy through ROS-Akt1-MTOR pathway in the HepG2 cells. [ABSTRACT FROM AUTHOR]

Abstract: This study investigated water‐insoluble corn flour (WICF) as a novel biosorbent for removing patulin from aqueous solutions, including contaminated fruit juices. Batch adsorption experiments were performed to assess the effect of granularity of WICF, adsorbent dosage, contact time, temperature, pH, and initial patulin concentration. The results showed that the maximum removal percentage of 92.21% was achieved at 37 °C, pH value of 4.0 in 100 µg/L patulin concentration for 25 hr. The adsorption data could be well fitted with the Freundlich isotherm model and the pseudo‐first order model. Thermodynamic parameters indicated that mycotoxin adsorption is an endothermic and spontaneous process. Subsequently, scanning electron microscopy images showed obvious morphological changes in WICF after adsorbing patulin. Fourier transform infrared spectroscopy spectroscopic evidence demonstrated that the main functional groups involved in the adsorption were O‐H, C = O and C‐O groups. Practical applications: Patulin was found as a contaminant in various fruit juices, which raised serious concerns about developing an effective and cheap adsorbent for detoxification of patulin toxin in fruit juice and their products. Water‐insoluble corn flour was applied to strengthen the control of patulin for the first time. Results of this study indicated that water‐insoluble corn flour has a strong ability to remove patulin from aqueous solutions and various fruit juices. Therefore, using water‐insoluble corn flour as an adsorbent and combining the filtration process could be potentially applied in fruit juice industry for detoxification of patulin. [ABSTRACT FROM AUTHOR]

This study reports for the first time in Serbia the occurrence of patulin in apple-based food and the risk assessment associated with patulin intake by infants and preschool children. In total, 214 samples of infant fruit juices (48), infant purée (66), and juices for children (small package with straw, 100) were collected over 3 years (2013–15) and analysed using HPLC with ultraviolet detection. Patulin was found in 43.8% of infant juices and 16.7% of infant purée, with all values below the legal limit of 10 μg kg−1 (maximum 8.3 and 7.7 μg kg−1, respectively). The proportion of contaminated samples among fruit juices for children was 43.0%, with the highest patulin concentration at 30.2 μg kg−1, not exceeding the maximum allowed level of 50 μg kg−1. Risk assessment of patulin intake by Serbian infants and preschool children, conducted by deterministic and probabilistic approaches, revealed a hazard quotient well below 1, indicating a tolerable exposure level and no health concern. [ABSTRACT FROM AUTHOR]

Patulin is a toxic metabolite of a number of fungi; its toxicity is serious and its contamination in food is a worldwide problem, especially in apple-based food. Effective control of patulin contamination strongly depends on reliable analytical methods. In this review, various analytical methods, especially those that have appeared in the last ten years, are summarized, including the highly reproducible chromatography and mass-spectrometry-based methods, highly selective sensor-based methods and indirect quantitative PCR methods. This review also summarizes the promising features of novel materials in sample preparation for patulin determination. [ABSTRACT FROM AUTHOR]

Background The present study demonstrates the antioxidant and hepatic protective effects of Green tea leaves (GTL). Methods The serum level of aspartate aminotransferase and alanine aminotransferase was analyzed. The liver antioxidant enzymes such as SOD, CAT, GPx, GR, GSH, lipid peroxidation and protein carbonyls, ROS content were estimated. The histology of liver tissue was observed and the protein expression of SOD, CAT, Caspase-3 and p53 was investigated by Western blotting. Results Effectiveness of GTL extract in preventing patulin induced liver damage showed significant reduction in serum ALT and AST to 19% and 85% respectively, the increase in antioxidant levels and lipid peroxidation products with patulin treatment were also reduced with GTL supplementation. The patulin induced increase in hepatic protein carbonyls was significantly reduced by 141–111% with 100 and 200 mg/kg b.wt GTL and in ROS was significantly reduced by 171–140% with 100 and 200 mg/kg b.wt GTL administration respectively. Also showed protection against hepatic tissue damage and protein expression in mice. Conclusion This study showed remarkable antioxidant and hepatic protective effects of GTL. [ABSTRACT FROM AUTHOR]

Mycotoxins are produced by different genera of fungi; mainly Aspergillus , Penicillium and Fusarium . The natural co-occurrence of beauvericin (BEA), patulin (PAT) and sterigmatocystin (STE) has been proved in feed and food commodities. This study investigates the cytotoxicity of individual and combined mycotoxins BEA, PAT and STE. The cytotoxicity on immortalized ovarian cells (CHO–K1) was evaluated using the MTT assay. After 24, 48 and 72 h, the IC 50 values were 2.9 μM for PAT and ranged from 10.7 to 2.2 μM and from 25.0 to 12.5 μM for BEA and STE, respectively. Cytotoxic interactions were assayed by the isobologram method, which provides a combination index (CI) value as a quantitative measure of the three mycotoxin interaction's degree. Binary and tertiary combinations showed a dose dependent effect. At low fraction affected, mycotoxin combinations were synergetic; whereas, at higher fraction affected, the combinations showed additive effect. Our results indicate that the co-occurrence of low concentrations of mycotoxin in food may increase their toxic effects. [ABSTRACT FROM AUTHOR]

Background: Patulin, a toxic mold metabolite, has been found as natural contaminant of processed fruits, most notably apples, apple juices and other apple-based products. A number of adverse health effects in humans and animals are associated with patulin intoxication. The current study was performed to analyse possible toxic effects of patulin on femoral bone microstructure in adult rabbits in detail. Fourteen clinically healthy four-month-old rabbits of both sexes (6 males and 8 females) were included in the study. Animals from the experimental groups (group E♂, n = 3; group E♀, n = 4) were injected intramuscularly with patulin at dose 10 μg/kg body weight two times a week for 28 days. The dose of patulin was estimated based on the maximum permitted level of patulin for apple products intended for infants and young children. Three males and four females without patulin administration served as controls (groups C♂ and C♀). Cortical bone thickness and qualitative and quantitative histological characteristics of compact bone tissue were investigated. Results: Intramuscular applications of patulin significantly increased the thickness of cortical bone in both sexes of rabbits. In patulin-exposed males, an absence of primary vascular longitudinal bone tissue near the endosteal border was observed, which could be associated with intensive bone remodeling. Femoral diaphyses of females displayed a lower number of secondary osteons in the middle part of the substantia compacta, and occurrence of the osteons near the periosteum. This could indicate alterations in bone turnover. Histomorphometrical evaluations showed significantly increased sizes of the primary osteons' vascular canals (P < 0.05) in males exposed to patulin possibly due to mycotoxin-induced increased levels of testosterone. Conclusions: This study demonstrates significant impact of intramuscular application of patulin on bone microstructure in adult rabbits. Moreover, we have found that the effects of patulin on qualitative and quantitative histological characteristics of compact bone are sex-dependent. [ABSTRACT FROM AUTHOR]

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Patulin (PAT) is a mycotoxin produced by certain species of Penicillium, Aspergillus, and Byssochlamys. Previous studies demonstrated its cytotoxic, genotoxic, and mutagenic effects in different cell lines. However, there is little information available concerning its toxic behavior in vivo . In the present study, we investigated PAT-induced hepatotoxicity and genotoxicity in mice. We also investigated the antioxidant and anti-genotoxicity efficiency of green tea polyphenols (GTP) against PAT-induced toxicity. We found that PAT-treatment induced serum alanine transaminase (ALT) and aspartate transaminase (AST) activities significantly. PAT-induced lipid peroxidation was confirmed with the elevation of thiobarbituric acid-reactive substances (TBARS). Moreover, the increasing of reactive oxygen species (ROS) and decreasing of GSH level implied its oxidative damage mechanism. In bone marrow cell, PAT was found to induce micronucleus and chromosomal aberration formation. In addition, our result suggested that GTP administration has dose-dependent antioxidative and antigenotoxic effect in against PAT-induced hepatotoxicity and genotoxicity. [ABSTRACT FROM AUTHOR]

Factors affecting ultraviolet ( UV) radiation-induced degradation of patulin in a malic acid model system, apple cider and apple juice were studied. First-order rate constants (cm2/J) in the model system were not significantly ( P > 0.05) affected by pH (3.0-3.6), initial patulin concentration (0-1000 μg/ L), glucose (0-10%), sucrose (0-10%) and ascorbic acid (0-100 mg/ L). Tannic acid (0-1 g/ L) reduced patulin degradation while fructose (0-10%) accelerated it. Although suspended particles yielding nephelometric turbidity units values between 0 and 100 did not affect patulin degradation, higher turbidity levels in unfiltered apple cider reduced degradation rates. Even in clarified apple juice, the UV dose required for 50% patulin reduction in apple juice (4.62 J/cm2) was higher than that required for a 5-log reduction of Escherichia coli O157: H7. Therefore, achieving efficient UV reduction of patulin in apple juice products, without quality losses, may require particulate filtration, phenolic adsorption treatments, and addition of compounds that accelerate patulin degradation. Practical Applications This study demonstrates that apple juice constituents affect patulin degradation during ultraviolet ( UV) processing. High turbidity levels in apple cider suggest that this technology may be impractical for this product. UV irradiation can be used to decrease patulin levels in clarified apple juice, although the required dose levels remain higher than that required for 5-log reduction of human pathogens. Filtration, adsorption treatments that lower levels of UV scattering and absorbing suspended particles and polyphenols can result in substantially greater patulin degradation during UV processing. Because fructose had an accelerating effect on patulin reduction in the model system studied, addition of high-fructose sweeteners to apple beverages prior to UV processing may result in further patulin reductions. [ABSTRACT FROM AUTHOR]

Abstract: A rapid, simple, improved method for the simultaneous determination of patulin (PAT) and 5-hydroxymethylfurfural (5-HMF) in fruit juices is described. The target compounds were extracted with ethyl acetate using vortex followed by high performance liquid chromatographic separation. PAT and 5-HMF were separated within 4 min using Poroshell C18 column with acetonitrile:water (1:9, v/v) as the mobile phase. Under the optimized conditions, the detection limits of PAT and 5-HMF were 0.25 and 0.46 ng mL−1, respectively while the quantification limits were 0.76 and 1.40 ng mL−1, respectively. The recoveries of PAT and 5-HMF at 50, 750 and 5000 ng g−1 ranged from 92.8 to 108%. The proposed method was applied to fivety-six fruit juices (apple, mango, pineapple, guava, lychee, tamarind, soursop and mixed fruit) and PAT was found in three samples ranging from 13.1–33.7 ug L−1. 5-HMF was found in all the samples ranging from 0.08 to 91.5 mg L−1. Liquid chromatography-tandem mass spectrometry with triple quadrupole analyzer was used to confirm the presence of 5-HMF and PAT in some of the contaminated samples. [Copyright &y& Elsevier]

Highlights: [•] There is higher number of patulin positives samples in organic apple-based products than in conventional ones. [•] The mean concentration of patulin was higher in organic than in conventional apple-based products. [•] All the estimated intakes of patulin calculated from the data of this study are below the PMTDI. [ABSTRACT FROM AUTHOR]

In this article, the authors discuss the role of mycotoxins during beer brewing and fermentation. They inform that levels of mycotoxins such as aflatoxins and ochratoxin A decrease to less than 20 percent of their initial concentration during the production of beer from barley and corn, and patulin mycotoxin is metabolized to a less toxic compound during the fermentation.

Patulin is a mycotoxin that contaminates pome fruits and derived products worldwide. Basidiomycete yeasts belonging to the subphylumPucciniomycotina have been identified to have the ability to degrade this molecule efficiently and have been explored through different approaches to understand this degradation process. In this study, Sporobolomyces sp. strain IAM 13481 was found to be able to degrade patulin to form two different breakdown products, desoxypatulinic acid and (Z)-ascladiol. To gain insight into the genetic basis of tolerance and degradation of patulin, more than 3,000 transfer DNA (T-DNA) insertional mutants were generated in strain IAM 13481 and screened for the inability to degrade patulin using a bioassay based on the sensitivity of Escherichia coli to patulin. Thirteen mutants showing reduced growth in the presence of patulin were isolated and further characterized. Genes disrupted in patulin-sensitive mutants included homologs of Saccharomyces cerevisiae YCK2, PAC2, DAL5, and VPS8. The patulin-sensitive mutants also exhibited hypersensitivity to reactive oxygen species as well as genotoxic and cell wall-destabilizing agents, suggesting that the inactivated genes are essential for tolerating and overcoming the initial toxicity of patulin. These results support a model whereby patulin degradation occurs through a multistep process that includes an initial tolerance to patulin that utilizes processes common to other external stresses, followed by two separate pathways for degradation. [ABSTRACT FROM AUTHOR]

Abstract: Citrinin (CTN) and patulin (PAT) are fungal secondary metabolites which are found in food and feed and showed organotoxicity in mature animals. In this study zebrafish embryos were applied to investigate the developmental toxicity of CTN and PAT on embryonic kidney. In the presence of CTN and PAT, the gross morphology of kidneys from embryos with green fluorescent kidney (wt1b:GFP) was not apparently altered. Histological analysis of CTN-treated embryos indicated cystic glomerular and tubular lesions; a disorganized arrangement of renal cells was also found in the PAT-treated group. From the view point of renal function, dextran clearance abilities of embryos exposed to CTN and PAT were significantly reduced. The damaged renal function caused by CTN could be partially rescued by the administration of pentoxifylline, suggesting the reduction of glomerular blood flow contributes to CTN-induced renal dysfunction. Additionally, CTN induced the expression of proinflammation genes, including COX2a, TNF-α and IL-1β, but failed to modify the levels and distribution of wt1a transcript and Na+/K+-ATPase protein. In summary, CTN and PAT caused profound nephrotoxicity in histological structure and biological function of zebrafish embryos; the inflammatory pathway and blood rheology may involve in CTN-induced renal impairment. [Copyright &y& Elsevier]

The fungal mycotoxin patulin is produced by several molds, especially by Aspergillus and Penicillium. The aim of this study was to clarify whether patulin causes alterations in plasma membrane permeability of Schizosaccharomyces pombe lead to cellular shrinkage charateristic to apoptosis or increases cell size indicating necrosis in cells. Transmission and scanning electronmicroscopy revealed that lower concentrations of patulin induced cellular shrinkage and blebbing, higher concentration caused expansion without cellular disruption. Large-scale morphological changes of individual cells were followed by time lapse video microscopy. Patulin caused the elongation and stickiness of cells or rounded up their shapes. To visualize chromatin structures of S. pombe nuclei upon patulin treatment, protoplasts were isolated from S. pombe and subjected to fluorescent microscopy. Chromatin changes in the presence of 50 μM patulin concentration were characterized by elongated nuclei containing sticky fibrillary chromatin and enlarged round shaped nuclei trapped at the fibrillary stage of chromatin condensation. Short (60 min) incubation of S. pombe cells in the presence of high (500 μM) patulin concentration generated patches of condensed chromatin bodies inside the nucleus and caused nuclear expansion, with the rest of chromatin remaining in fibrillary form. Longer (90 min, 500 μM) incubation resulted in fewer highly condensed chromatin patches and in nuclear fragmentation. Although, high patulin concentration increased the size of S. pombe size, it did not lead to necrotic explosion of cells, neither did the fragmented nuclei resemble apoptotic bodies that would have indicated programmed cell death. All these morphological changes and the high rate of cell survival point to rapid adaptation and mixed type of fungistatic effects. (© 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim) [ABSTRACT FROM AUTHOR]

Abstract: Apple juices and purees represent categories widely consumed by whole population and above all children. Patulin (PAT) is a mycotoxin known for its acute and chronic effects in animals. Several studies indicate there is a risk associated to the PAT intake, through the consumption of purees and apple juices. In this study, apple juice and puree were prepared and artificially contaminated with PAT at 50μg/kg and submitted to a thermal treatment simulating pasteurization to evaluate PAT’s reduction. In a second phase of the work, apple products samples (n =7) included juices, nectars and purees belonging to different commercial brands were collected, artificially contaminated with PAT at 50μg/L (limit established for PAT in juices) and 25μg/kg (limit established for PAT in purees), digested with an in vitro gastrointestinal protocol and bioaccessibility values (%) were calculated. After thermal treatment, the PAT’s loss evidenced in purees and juices was of 1.41±0.52% and 62.62±2.53% respectively. Related to the bioaccessibility data, two juices with pulp showed values of 70.89±4.93 and 67.30±10.76%; two purees showed levels of 58.15±5.50 and 55.69±4.73%, whereas nectar and two clarified juice showed percentages of 38.88±2.42, 28.59±0.46 and 25.28±0.61%, respectively. [Copyright &y& Elsevier]

Abstract: Patulin is a mycotoxin and its contamination of food has been reported to cause gastrointestinal inflammation, ulcers, and bleeding. The toxicity of patulin is thought to be due to the destruction of tight junctions (TJs) in gastrointestinal tissues. However, the precise mechanism has not been clarified. Here, we investigated the phosphorylation of TJ components. The transepithelial electrical resistance (TER) of Caco-2 human colon cancer cells decreased gradually during the first 24h of treatment with 50μM patulin. Immunofluorescence microscopy showed that the TJ proteins ZO-1 and claudin-4, but not occludin, had decreased after 24h and decreased from the cell–cell contact regions of TJs after 48h of patulin treatment. Western blotting showed that the level of ZO-1 decreased after 48h of patulin treatment, but the levels of claudin-4 and occludin remained at the initial level until 72h. Phosphorylation of ZO-1 was detected by 24h and increased markedly after 72h of patulin treatment. However, phosphorylation of claudin-4 and occludin was not detected by probing with anti-phosphotyrosine antibody. Immunoprecipitation showed that interaction of ZO-1 with claudin-4 had decreased after 48h and was completely absent after 72h. These results suggest that phosphorylation caused the degradation of ZO-1 protein and the decrease in TER induced by patulin treatment of Caco-2 cells. [Copyright &y& Elsevier]

Abstract: Although the production of patulin in apple fruits is mainly by Penicillium expansum, there is no information on the ability of heat resistant moulds that may survive pasteurization to produce this mycotoxin in juice packages during storage and distribution. In this study, the production of patulin by Byssochlamys spp (Byssochlamys nivea FRR 4421, B. nivea ATCC 24008 and Byssochlamys fulva IOC 4518) in cloudy and clarified apple juices packaged in laminated paperboard packages or in polyethylene terephthalate bottles (PET) and stored at both 21°C and 30°C, was investigated. The three Byssochlamys strains were able to produce patulin in both cloudy and clarified apple juices. Overall, the lower the storage temperature, the lower the patulin levels and mycelium dry weight in the apple juices (p<0.05). The greatest variations in pH and °Brix were observed in the juices from which the greatest mycelium dry weights were recovered. The maximum levels of patulin recovered from the juices were ca. 150μg/kg at 21°C and 220μg/kg at 30°C. HPLC-UV, HPCL-DAD and mass spectrometry analyses confirmed the ability of B. fulva IOC 4518 to produce patulin. Due to the heat resistance of B. nivea and B. fulva and their ability to produce patulin either in PET bottles or in laminated paperboard packages, the control of contamination and the incidence of these fungi should be a matter of concern for food safety. Control measures taken by juice industries must also focus on controlling the ascospores of heat resistant moulds. [ABSTRACT FROM AUTHOR]

Abstract: Patulin is a toxic metabolite produced by various species of Penicillium, Aspergillus and Byssochlamys. In the present study, its effects on the plasma membrane of fission yeast Schizosaccharomyces pombe were investigated. The phase-transition temperature (G) of untreated cells, measured by electron paramagnetic resonance spectrometry proved to be 14.1°C. Treatment of cells for 20min with 50, 500, or 1000μM patulin resulted in a decrease of the G value of the plasma membrane to 13.9, 10.1 or 8.7°C, respectively. This change in the transition temperature was accompanied by the loss of compounds absorbing light at 260nm. Treatment of cells with 50, 500 or 1000μM patulin for 20min induced the efflux of 25%, 30.5% or 34%, respectively, of these compounds. Besides its cytotoxic effects an adaptation process was observed. This is the first study to describe the direct interaction of patulin with the plasma membrane, a process which could definitely contribute to the adverse toxic effects induced by patulin. [Copyright &y& Elsevier]

Patulin is a mycotoxin produced by several Penicillium, Aspergillus, and Byssachlamys species growing on food products. In this study, we investigated the effects of patulin on the thymus of growing male rats aged five to six weeks. The rats were receiving it orally at a dose of 0.1 mg kg-1 bw a day for either 60 or 90 days. At the end of the experiment, the thymus was examined for histopathology by light microscopy and for epidermal growth factor (EGF) and its receptor (EGFR) by immunolocalisation. For morphometry we used the Bs200prop program to analyse images obtained with the Olympus BX51 light microscope. Cell ultrastructure was studied by electron microscopy. In rats treated with patulin, the thymus showed haemorrhage, plasma cell hyperplasia, a dilation and fibrosis in the cortex, enlarged interstitial tissue between the thymic lobules, enlarged fat tissue, thinning of the cortex, and blurring of the cortico-medullary demarcation. Electron microscopy showed signs of cell destruction, abnormalities of the nucleus and organelles, and loss of mitochondrial cristae. However, no differences were observed in thymus EGF and EGFR immunoreactivity between treated and control rats. [ABSTRACT FROM AUTHOR]

This study was conducted to assess patulin exposure in the Catalonian population. Patulin levels were determined in 161 apple juice samples, 77 solid apple-based food samples and 146 apple-based baby food samples obtained from six hypermarkets and supermarkets from twelve main cities of Catalonia, Spain. Patulin was analysed by a well-established validated method involving ethyl acetate extraction and direct analysis by high-performance liquid chromatography (HPLC) with ultraviolet light detection. Mean patulin levels for positive samples in apple juice, solid apple-based food and apple-based baby food were 8.05, 13.54 and 7.12 µg kg-1, respectively. No samples exceeded the maximum permitted levels established by European Union regulation. Dietary intake was separately assessed for babies, infants and adults through a Food Frequency Questionnaire developed from 1056 individuals from Catalonia. Babies were the main group exposed to patulin, however no risk was detected at these levels of contamination. Adults and infants consumers were far from risk levels. Another approach to determine estimated exposure was conducted through Monte Carlo simulation that distinguishes variability in exposures from uncertainty of distributional parameter estimates. [ABSTRACT FROM AUTHOR]

This study examined the potential for patulin production in six different varieties of apples (Red Delicious, Golden Supreme, Gala, Fuji, Empire, and Mclntosh) inoculated with Penicillium expansum spores and stored at two different temperatures (11 and 20.5°C). Samples for patulin analysis were randomly taken from apples stored at different times, ranging from 21 to 93 days. While patulin was produced at both storage temperatures, apples incubated at 20.5°C yielded significantly higher patulin concentrations than did those incubated at 11°C. All apple varieties showed mold spoilage at both temperatures, except Red Delicious and Empire. A total of 44% of the samples analyzed showed patulin concentrations above the U.S. Food and Drug Administration regulatory limit (50 ppb). The highest patulin productions occurred in Golden Supreme (54,221 ppb) and Mclntosh (52,131 and 48,457 ppb) varieties. Our results showed that careful culling of apples is essential for high juice quality, since high patulin levels in some apples varieties could result in a level greater than 50 ppb of this mycotoxin in the finished juice or cider, even when only one contaminated apple occurs in 1,000 apples. [ABSTRACT FROM AUTHOR]

The article focuses on the study of molecular cloning and characterization of patulin in Aspergillus clavatus. Patulin is a tetraketide mycotoxin made by various fungal species. The study shows the major involvement of cytochrome P450 monoxygenases in the hydroxylation of patulin. It identifies two cytochrome P45o genes in the DNA sequence of Aspergillus clavatus species. Both mRNAs are expressed at the patulin production. The study recommends the major involvement of cytochrome in patulin biosynthesis.

Patulin is a marker of quality in the apple and apple juice industry and due to the potential risk for human health, reliable and potential methods for extracting patulin from a sample are therefore needed. In this study, the three methods with liquid–liquid extraction, matrix solid-phase dispersion (MSPD) and with solid-phase extraction (SPE) were studied for extracting patulin from different apple products. Result showed that for AJC and apple sample, MSPD method is most suitable for extracting patulin among the three methods. The recovery rates of AJC and apple sample were 80.35–114.46 and 79.68–94.32%, respectively, the coefficient variations were 3.18–4.90%; For dilute juice, SPE procedure is suitable for analysis of patulin and the recovery rates were and 85.35–90.14%. [ABSTRACT FROM AUTHOR]

Abstract: A survey was conducted to determine levels and dietary intake of Patulin (PAT) from apple juices consumed in Spain. One hundred samples of apple juice were bought from distinct supermarkets. PAT was extracted by a liquid–liquid extraction technique and analysed with a micellar electrokinetic chromatography (MEKC) method. 66% of the samples contained PAT over the limit of detection of the method (0.7μgL−1). The PAT apple juice mean and median levels obtained were 19.4 and 4.8μgL−1, respectively, in a range between 0.7 and 118.7μgL−1. In 11% of the samples, PAT contamination exceeded the maximum permitted level of 50μgL−1 established by the EU regulation. In Spain, no significant variations were observed with respect to data published 15 years ago. Intakes were calculated for average consumers among adults, children and babies and then compared with the provisional maximum tolerable daily intake (PMTDI) proposed by JECFA in 1995. Estimated daily intake of PAT in this study represents 14%, 39% and 63% of the PMTDI, respectively. [Copyright &y& Elsevier]

Aims: Because of the lack of a standard method, the aim of this work is to evaluate the suitability of the broth microdilution method CLSI M38-A in determining the resistance level of some Penicillium expansum isolates to thiabendazole (TBZ). The ability of the isolates to produce patulin (PAT) and citrinin (CIT) has been also assessed. Methods and Results: Penicillium expansum isolates (128) were assayed (apples, pears, grapes and five reference strains). It was observed that 69·4% of the strains isolated from apples and pears were resistant to TBZ. Sensitive isolates were inhibited at 0·25–0·5 μg ml−1 whilst resistant isolates still grew at 512 μg ml−1. PAT was produced by all P. expansum isolates. CIT was detected in 98·8% of TBZ-resistant isolates and in 89·1% of the TBZ-sensitive isolates. Conclusions: The preliminary screening method combined with the adaptation of the method CLSI M38-A, can be a good strategy to be used in assessing the in vitro activity of TBZ against a large number of isolates. Significance and Impact of the Study: The proposed methodology can be a contribution to the standardization of susceptibility tests to fungicides against P. expansum. [ABSTRACT FROM AUTHOR]

The effectiveness of seven phenolic compounds (esculetin, ferulic acid, quercetin, resveratrol, scopoletin, scoparone and umbelliferone) in controlling Penicillium expansum growth and patulin accumulation was evaluated by both in vitro and in vivo studies. The in vitro screening showed that quercetin and umbelliferone were the most effective compounds in controlling Penicillium growth and patulin accumulation, respectively. Quercetin and umbelliferone, resulted to be effective also in the in vivo screening, were further tested (alone and in combination) on Granny Smith and Golden Delicious apples. The efficacy in controlling Penicillium growth was better expressed on Golden Delicious than on Granny Smith apples, with quercetin providing a better control of both incidence of decay and disease severity as compared to umbelliferone. Both compounds exerted a considerable control of patulin accumulation on the two apple cultivars. Quercetin and umbelliferone can be considered as natural compounds to be used as alternative strategy to chemical fungicides in postharvest control of P. expansum infections on apples. [ABSTRACT FROM AUTHOR]

Abstract: Patulin (PAT) is a fungal secondary metabolite that exhibits potential cellular and animal toxicities. In this study, human promyelocytic leukemia (HL-60) cells were used to elucidate the mechanism and death mode associated with PAT. Morphological evidence of apoptosis, including membrane blebbing, nuclei fragmentation and DNA laddering formation was clearly observed 6h after exposure to PAT. The results of Western blotting indicated that PAT activated various processed caspases, and cleaved DFF45 and poly (ADP-ribose) polymerase (PARP) in a dose-dependent manner; it also induced a time-dependent increase in caspase 3 and 9 catalytic activities. The apoptosis mediated by PAT in HL-60 was accompanied with cytochrome c release from mitochondria and Bcl-2 expression decrease. The presence of thiol-containing compounds with PAT dramatically reduced the caspase 3 activity that was triggered by PAT; the addition of antioxidants, including mannitol and Tiron, had a similar effect. However, the suppression of p53 protein expression by RNA interference (RNAi) in human embryonic kidney (HEK293) cells did not significantly modify PAT-elicited caspase 3 activity. These findings suggest that PAT-induced apoptosis is mediated through the mitochondrial pathway without the involvement of p53; the interaction with sulfhydryl groups of macromolecules by PAT and the subsequent generation of reactive oxygen species (ROS) plays a primary role in the apoptotic process. [Copyright &y& Elsevier]

Fifty samples of apple juices were investigated for patulin levels using HPLC with UV detection. It was found that 38 (76? samples contained no detectable patulin (LOD = 7.5 µg l-1), but some samples had high patulin levels (845 µg l-1) compared to other branded mixed juices (21-70 µg l-1) and apple juices procured from local vendors (191 µg l-1). The survey provides evidence of a need to educate the apple-juice industry in the use of good quality apples for juice production. In addition, the levels of patulin found showed a saturation of the provisional maximum tolerable daily intake (PMTDI) based on intake figures in terms of an average and 90th percentile of 27-60% and 107-190%, respectively, in different age groups. This monitoring study may help in formulating guidelines for patulin levels, which are not yet implemented by Prevention of Food Adulteration (PFA) Act of India. [ABSTRACT FROM AUTHOR]

This study investigated the effect of patulin and penicillic acid, two known quorum-sensing inhibitors, and the common biocide ethylenediaminetetraacetic acid (EDTA) on the biofilm formation and auto-inducer (AI)-2 production of three isolates from dental unit water lines, Klebsiella sp., Bacillus subtilis and Bacillus cereus. Penicillic acid on its own had no effect on the biofilm formation of all isolates, whereas in combination with EDTA, it enhanced biofilm formation significantly in Klebsiella sp. and B. cereus. EDTA at concentrations greater than 10 μM promoted biofilm formation in B. cereus and B. subtilis. Patulin was found to promote biofilm formation in B. cereus up to 25 μM. A significant increase in biofilm formation was observed in B. cereus and B. subtilis at concentrations greater than 10 μM of patulin when combined with EDTA. The Vibrio harveyi BB170 AI-2 bioassay showed a positive response for Klebsiella sp. AI-2 production with a maximum fold induction at the late exponential growth phase. Addition of glucose prolonged the AI-2 production phase considerably. No significant effect of patulin, penicillic acid alone as well as in combination with EDTA was observed on AI-2 production by Klebsiella sp. The findings have important implications for the design of biofilm prevention and eradication strategies. [ABSTRACT FROM AUTHOR]

Aims: To assess the ability of fungi isolated from grapes to produce patulin and citrinin. Methods and Results: A total of 446 Aspergillus isolates belonging to 20 species and 101 Penicillium isolates were inoculated in Czapek yeast extract agar and yeast extract sucrose agar and incubated for 7 days at 25°C. Extracts were analysed for patulin and citrinin by thin-layer chromatography. None of the isolates of Aspergillus spp. produced either patulin or citrinin. Patulin was produced by three isolates of Penicillium expansum and two of Penicillium griseofulvum. Citrinin was produced by five isolates of P. expansum, two of Penicillium citrinum and one of Penicillium verrucosum. Conclusions: Our results show that the Aspergillus and Penicillium species commonly isolated from grapes are not a source of the mycotoxins, patulin and citrinin. Significance and Impact of the Study: The possibility of co-occurrence of patulin and citrinin with ochratoxin A in grapes and grape products remain low, owing to the low frequency of isolation of potentially producing species. [ABSTRACT FROM AUTHOR]

Abstract: A high performance liquid chromatography with a diode-array detector (HPLC-DAD) and a gas chromatography with a mass spectrometer (GC-MSD) are described for the determination of patulin (PAT) in apple juice. The limits of detection (DL) and quantification (QL) for the HPLC-DAD and GC-MSD method were found to be (DL=0.23μgkg−1 QL=1.2μgkg−1) and (DL=5.8μgkg−1 and QL=13.8μgkg−1), respectively. The recovery factors for HPLC-DAD and GC-MSD were found to be 99.5% (RSD%=0.73) and 41% (RSD%=10.03), respectively. The HPLC-DAD method was used to determine the occurrence of PAT in 90 samples of fruit juices. Results revealed the presence of PAT in 100% of the samples examined. The mean values of PAT in concentrated fruit juices and in the commercial fruit juices collected from the Greek market were found to be 10.54μg PAT kg−1 and 5.57μg PAT kg−1 juice, respectively. The most contaminated samples were four concentrated juices ranging from 18.10μg PAT kg−1 to 36.8μg PAT kg−1 juice. The daily exposure to patulin for the consumers of all ages in Greece, is ranging from 0.008μg PAT kg−1 bw to 0.1μg PAT kg−1 bw if the daily intake of fruit juices is from 0.1 to 0.5kg. With the exception to the most contaminated sample, the daily exposure due to the samples examined, is below the provisional maximum tolerable daily intake for PAT (0.4μg PAT kg−1 bw). [Copyright &y& Elsevier]

Patulin is a frequent contaminant of moldy and rotten apples and apple products. The aim of this study was to evaluate patulin contamination in 58 apple juices collected from a retail market in Mashhad during winter and spring of 2006. Samples were assayed for patulin by high-performance liquid chromatography. Fifty-four samples were positive for patulin at levels that ranged from 10.5 to 121.8 µg/L, and six samples had patulin levels higher than 50 µg/L. The overall mean of patulin concentration was 29.2 ± 19.5 µg/L. Forty-eight samples had patulin concentration between 5 and 50 µg/L. Although the mean concentration of patulin samples was lower than Iranian maximum tolerated level of 50 µg/L, contamination of 10% of the samples at levels higher than 50 µg/L indicated the need for improving production techniques by the industry. PRACTICAL APPLICATIONS Studies have shown that the concentration of patulin may exceed the determined limits in apple juice and in other fruit products. Its presence can be a potential threat to the health of consumers, particularly children. The results may help us in understanding what should be the level of patulin in apple juice. The awareness of the apple industry of patulin contamination in fruit and the implementation of improved techniques for the production of apple products with reduced patulin concentrations have contributed to the quality of apple juice that are available on the Iranian market. [ABSTRACT FROM AUTHOR]

Patulin, a food toxin that can be present in pear juice and especially in apple juice can be destroyed by the action of ozone. In this paper, we show by HPLC-DAD chromatography that patulin, both in a water solution or in apple juice, can be destroyed by the action of ozone. [ABSTRACT FROM AUTHOR]

Abstract: Aim of the present study was to investigate the detoxification of two abundant mycotoxins, namely ochratoxin A (OTA) and patulin (PAT) which are frequently found in human foods, by lactic acid bacteria. The removal of the two mycotoxins from liquid medium by thirty different LAB strains was analyzed in a screening trial by the use of HPLC coupled with UV- or fluorescence detection. Two highly effective strains were identified; Lactobacillus acidophilus VM 20 caused a decrease of OTA by ⩾95% and Bifidobacterium animalis VM 12 reduced PAT levels by 80%. Subsequently experiments showed that the binding of these compounds depends on different parameters, i.e. the concentration of toxins, the cell density, the pH-value and on the viability of the bacteria. To proof that the decrease of the toxins by LAB from liquid medium results in a reduction of their toxic properties, micronucleus (MCN) assays were conducted with a human derived hepatoma cell line (HepG2). Indeed, a substantial decrease (39–59%) of OTA and PAT induced MCN formation was observed with the most effective strains detected in the chemical analyses. Furthermore, also the inhibition of the cell division rates by the toxins was significantly reduced. These findings indicate that certain LAB strains are able to detoxify the two toxins and may be useful to protect humans and/or animals against the adverse health effects of these compounds. [Copyright &y& Elsevier]

This article discusses the effects of the mycotoxin patulin on the development of human peripheral blood mononuclear cells (PBMC). Effects of patulin on PBMCs were evaluated by cytokine ELISAs, proliferation, and viability assays. Patulin decreased the levels of several cytokines, but not all of them, and depleted intracellular glutathione. They argued that patulin modulates PBMCs and regulated cytokine secretion patterns by glutathione depletion. This may influence the balance between Th1 and Th2 cells which could have implications for allergic diseases.

Abstract: Penicillium expansum causes blue mould rot, a serious post-harvest disease of apples, and is the main producer of the mycotoxin patulin. The present study aimed to determine the influence of storage conditions (i.e. temperature and O2 level) on growth and patulin production by different P. expansum strains on a simulation medium and on apples. Growth was strongly influenced by the temperature, while the used atmosphere (20, 3, and 1% O2; <1% CO2) had no effect. Optimal growth was observed at 25 °C for every strain tested. Patulin production was stimulated when the temperature decreased (from 20 to 10 or 4 °C), while a further decrease of the temperature to 1 °C caused a reduction in patulin production. The temperature at which the stimulation was changed into suppression was strain dependent. Similar results were observed for the O2 level. A reduction of the O2 level from 20 to 3% O2 could stimulate or suppress patulin production depending on the strain, while a clear decrease of the patulin production was observed when the O2 level was reduced from 3 to 1%. These results show that the induction of limited stress to the fungus, such as lowering the temperature or lowering the O2 levels stimulates patulin production. However, the combination of different stress conditions (e.g. low temperature and low O2) will result in a reduced formation of the toxin. The combination of stress conditions, at which the transition from stimulation to suppression is observed, is strain dependent. Moreover, patulin production is characterized by a high natural variability. The presented results show that the temperature and O2 level has to be as low as possible during the storage of apples in order to suppress patulin production and to guarantee food safety. [Copyright &y& Elsevier]

Abstract: The aim of this study was to assess the opportunities of Penicillium expansum to develop and produce patulin in apples under two different CA storage methods (LOW and U-LOW) at 1°C. Differences in lesion diameter and patulin accumulation depending on CO2 and O2 partial pressure were studied. Further apple rot and patulin production during a three days post-storing stage at 20°C was also monitored so that effect of further storage at room temperature could be assesed. Two lots of apples of Golden variety with different ripeness degrees were used. Half of each lot was fungicide treated. Apples were inoculated with patulin producer P. expansum strains and stored at 1°C for either two month or 2.5 months at both LOW and U-LOW conditions. The extent of lesions and patulin accumulation both at the end of CA cold storage and after three days at 20°C were assessed. CA storage conditions had strong significance in P. expansum growth on apples and factors such as fruit ripeness, fungicide treatment and time at the storage room had significant influence. In general, bigger lesions were observed under U-LOW than under LOW conditions, lesions being similar or bigger when increasing the storage time from 2 to 2.5 months. P. expansum grew faster in riper apples, although fungicide application was clearly more effective for ripe rather than for underripe apples. Although lesions were evident after both storage conditions, no patulin was detected. Increase of lesion when fruits were subsequently stored at 20°C was evident in all cases and patulin was detected at this moment. No differences in patulin content were found at this stage between LOW and U-LOW stored apples. [Copyright &y& Elsevier]

Patulin is a mycotoxin produced by species of Penicillium, Aspergillus and Byssochylamys. Several Scientific Committees classify patulin as mutagenic, embryotoxic and immunotoxic. It has been found as a natural contaminant of processed apple products and its presence may be indicative of the quality of fruit used in production. In this work, a method for the analysis of patulin is described, based on a simple liquid-liquid extraction with acetonitrile; patulin is analyzed using liquid chromatography with UV detection. Patulin identity was confirmed by GC-MS after its reaction with N-methyl-N-(trimethylsilyl)trifluoroacetamide. Fifty-three apple-containing products were analyzed and patulin was detected in 14 samples in a range 1.5-50.9 µg l-1; six of which were above the maximum permitted level of the European Union. Based on these results and juice consumption by the Spanish adult population, patulin estimated intake was 0.42 ng kg-1 body weight per day. [ABSTRACT FROM AUTHOR]

Abstract: The objective of the present study was to evaluate the patulin exposure of children consuming organic, handcrafted or conventional apple juice through a probabilistic approach and to evaluate the effectiveness of several risk management options aiming to reduce the risk for children due to patulin exposure. However, a large part of the data on patulin contamination of apple juice fell under the limit of detection (LOD). Different methods were tested to deal with these so-called left censored data and a uniform distribution with uncertain bounds was selected to handle this censorship. Variability and uncertainty assessment of patulin exposure showed that 0.9% [90% confidence interval (CI): 0.3–1.8%] of the children consuming only organic apple juice exceed the tolerable daily intake (TDI). For consumers of conventional and handcrafted apple juice this was respectively 0.1% [90% CI: 0–0.3%] and 0% [90% CI: 0–0.2%]. Reduction of the patulin contamination in apple juice to concentrations below 25μg/kg reduced the percentage of the children exceeding the TDI to 0% [90%CI: 0–0.2%] for organic apple juice. Reduction of the apple juice consumption was less effective than a reduction of the patulin concentration in apple juice and is only useful when the patulin concentration of apple juice is below 25μg/kg. [Copyright &y& Elsevier]

A study of apple juice products sold in Italy and South Africa was initially carried out on 20 samples bought in Cesena, Italy, and Tygerberg in Cape Town, South Africa. The samples were bought at random and analysed for patulin contamination. All 12 of the Italian samples had no detectable levels of patulin, except one, which was just slightly above the lowest regulatory limit of 10 ng ml-1. On the other hand, five of the eight South African samples were all contaminated with patulin levels above 10 ng ml-1, with one showing a concentration of 75 ng ml-1, well above the highest regulated limit of 50 ng ml-1. This latter result led to a more targeted investigation with 14 samples being purchased in the low-income areas of Tygerberg where the initial samples were sourced. These samples confirmed that there might be a problem of mycotoxin contamination in apple juices products sold to low-income consumers because half of the samples showed patulin contamination of which four had levels well above the acceptable limits. This is the first study in South Africa to look at apple juice products in low-income areas and it points to a need to intervene and introduce quality systems in the supply chain of the manufacture and packaging of apple juice products by independent small business. [ABSTRACT FROM AUTHOR]

Patulin (PAT), a mycotoxin mainly produced by Penicillium and Aspergillus, is found in various foods and feeds. In the present study, its effects on oxidative stress in various mammalian cell lines were investigated. When cell-permeating fluorescent dyes were used as indicators of the generation of reactive oxygen species (ROS), we found that PAT treatment directly increased intracellular oxidative stress in human embryonic kidney (HEK293) and human promyelocytic leukemia (HL-60) cells. Lipid peroxidation levels were also significantly increased in HL-60 cells and mouse kidney homogenates treated with PAT. Suppression of CuZn–superoxide dismutase (SOD) expression in mammalian cells by small interfering RNA resulted in an increase in PAT-mediated membrane damage, while overexpression of human CuZn-SOD or catalase led to a reduction in damage, indicating the involvement of ROS in PAT toxicity. Pretreatment of HEK293 cells with Tiron, a free radical scavenger, reduced the phosphorylation levels of extracellular signal–regulated kinase (ERK) 1/2 elicited by PAT. The ERK1/2 signaling pathway inhibitor, U0126, also significantly decreased the levels of ROS associated with PAT treatment. These findings indicate that PAT treatment results in the ROS production in mammalian cells, and ROS partially contributes to PAT-induced cytotoxicity. Activation of ERK1/2 signaling pathway is correlated with PAT-mediated ROS. [ABSTRACT FROM AUTHOR]

A bacterium isolated from patulin-contaminated apples was capable of degrading patulin to a less-toxic compound, asciadiol. The bacterium was identified as Gluconobacter oxydans by 16S rRNA gene sequencing, whereas ascladiol was identified by liquid chromatography-tandem mass spectrometry and proton and carbon nuclear magnetic resonance. Degradation of up to 96% of patulin was observed in apple juices containing up to 800 µg/ml of patulin and incubated with G. oxydans. [ABSTRACT FROM AUTHOR]

Patulin is a toxic secondary metabolite of a number of fungal species belonging to the genera Penicillum and Aspergillus. It has been mainly isolated from apples and apple products contaminated with the common storage-rot fungus of apples, Penicillum expansum, but it has also been extracted from rotten fruits, moldy feeds, and stored cheese. Human exposure to patulin can lead to serious health problems, and according to a long-term investigation in rats, the World Health Organization has set a tolerable weekly intake of 7 ppb body weight. The content of patulin in foods has been restricted to 50 ppb in many countries. Conventional analytical detection methods involve chromatographic analyses, such as HPLC, GC, and, more recently, techniques such as LC/MS and GC/MS. However, extensive protocols of sample cleanup are required prior to the analysis, and to accomplish it, expensive analytical instrumentation is necessary. An inimunochemical analytical method, based on highly specific antigen-antibody interactions, would be desirable, offering several advantages compared to conventional techniques, i.e., low cost per sample, high selectivity, high sensitivity, and high throughput. In this paper, the synthesis of two new derivatives of patulin is described, along with their conjugation to the bovine serum albumin for the production of polyclonal antibodies. Finally, a fluorescence competitive immunoassay was developed for the on-line detection of patulin. [ABSTRACT FROM AUTHOR]

Aims: To investigate the effects of trans-2-hexenal on blue mould disease, patulin content and fruit quality in ‘Conference’ pears. Methods and Results: Fruits, wounded and inoculated with Penicillium expansum or non-inoculated, were exposed to trans-2-hexenal vapour treatment (12·5 μl l−1) at 20°C. A greater reduction of decay was obtained by treatment application 24 or 48 h after inoculation, in contrast trans-2-hexenal application 2 h after inoculation was ineffective. Fruit storage temperature (−1°C) after treatment did not affect the antifungal activity. Although 2-h exposure to trans-2-hexenal was effective in reducing blue mould, an exposure of at least 8 h was required to reduce fruit patulin content. Treatments did not affect fruit physical–chemical characteristics. After 6 days at 20°C following exposure, trans-2-hexenal residue in treated fruits was less than the natural content of the compound in unripe fruits. Conclusions: trans-2-Hexenal treatment is effective in the reduction of blue mould infections and patulin content in Conference pears when applied 24–48 h after pathogen inoculation. Significance and Impact of the Study: trans-2-Hexenal could be a natural alternative to fungicides in the control of P. expansum infections. Further work is needed to study the methods and conditions avoiding the persistence of off-odours and off-flavours in pears after their exposure to trans-2-hexenal vapours. [ABSTRACT FROM AUTHOR]

Patulin (PAT), a mycotoxin mainly produced by Penicillium and Aspergillus, is frequently detected in moldy fruits and fruit products. Exposure of human embryonic kidney (HEK293) cells to PAT led to a dose- and time-dependent increase in the phosphorylation of two major mitogen-activated protein kinases (MAPKs), p38 kinase and c-Jun N-terminal kinase (JNK). The phosphorylated forms of MAPK kinase 4 (MKK4), c-Jun, and ATF-2 were also seen in PAT-treated cultures. The cell death caused by PAT was significantly reduced by the p38 kinase inhibitor, SB203580, but not by the JNK inhibitor, SP600125. Neither p38 kinase nor JNK played a role in the PAT-induced DNA damage. In PAT-treated cells, inactivation of double-stranded RNA-activated protein kinase R (PKR) by the inhibitor, adenine, markedly suppressed JNK and ERK phosphorylation. Treatment of HEK293 cells with PAT-cysteine adduct, a chemical derivative of PAT, showed no effect on MAPK signaling pathways, cell viability, or DNA integrity. These results indicate that PAT causes rapid activation of p38 kinase and JNK in HEK293 cells, but only the p38 kinase signaling pathway contributes to the PAT-induced cell death. PKR also plays a role in PAT-mediated MAPK activation. [ABSTRACT FROM AUTHOR]

Patulin is a mycotoxin produced by fungal species that frequently grow on fruit and vegetables. It presents risks, particularly for children consuming compotes and fruit juices. Thus, it is important to have methods such as immunoassays to screen a large number of samples. In the relevant literature, previous studies on the production of antibodies against patulin derivatives described qualitative tests for a patulin derivative or showed slight responses. The present study reinvestigated the production of polyclonal antibodies against patulin and their purification since crude antiserum could react non-specifically in immunoassays. Patulin-hemiglutarate was synthesized and conjugated to bovine serum albumin as the immunogen for the immunization of five New Zealand white rabbits. The immunoglobulin G (IgG) fraction was isolated twice by affinity chromatography using Sepharose-LS® gel and recombinant G-protein. Classic affinity chromatography using Sepharose-LS gel was unable to eliminate serum albumin from the IgG fraction and the use of recombinant G-protein was efficient to isolate the purified IgG. Titres and specificity were determined by indirect competitive enzyme-linked immunosorbent assay. Patulin-hemiglutarate-ovalbumin gave complete displacement, while patulin displaced 30% of bound antibodies. Thus, a fraction of the antibodies are specific for free patulin. The non-specific binding increased with patulin concentrations. The electrophilic properties of patulin might also induce intermolecular cross-links in vitro that hinder the possibility of responses displacement when free patulin is used. [ABSTRACT FROM AUTHOR]

The occurrence of patulin was investigated in 100 conventional and 69 organic fruity foodstuffs samples commercially available in Italy by using an HPLC method with a limit of quantification of 0.5?µg?kg -1 . Patulin was detected in 26 (26%) conventional and 31 (45%) organic products with a significantly higher ( p <0.01) mean concentration in the organic products (4.78 vs. 1.15?µg?kg -1 ). Mean patulin concentrations in conventional apple juices, pear juices, other juices and fruits purees were 3.14, 0.22, 0.19, 0.11?µg?kg -1 , respectively, and 7.11, 11.46, 2.10, 0.18?µg?kg -1 in the relevant organic products. Four samples of juices (one conventional and two organic apple, and one organic pear) contained patulin at concentrations above the limit of 50?µg?kg -1 , four at concentrations between 10 and 25?µg kg -1 , and the remaining ones below 10?µg?kg -1 . Patulin was detected (<1?µg?kg -1 ) in only three of the 23 fruity baby food samples tested (homogenized fruits, 11 conventional and 12 organic). Based on the available data on Italian intakes of fruit juices, the estimated daily intakes of patulin, were 0.38 and 1.57?ng?kg -1 body weight (bw) from conventional and organic products, respectively. Estimated daily intakes of patulin for children were higher, 3.41?ng?kg -1 bw from conventional and 14.17?ng?kg -1 bw from organic products, but largely below the provisional maximum tolerable daily intake (PMTDI) of 400?ng?kg -1 bw. Patulin was also found in two samples of organic apple vinegar (<5?µg?kg -1 ) and in fresh apples with rotten spots (12 out of 24 samples) with maximum levels at 16?402 and 44?572?µg?kg -1 for conventional and organic apples, respectively. [ABSTRACT FROM AUTHOR]