Study of thermal resistance and in vitro bioaccessibility of patulin from artificially contaminated apple products

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 50 μg/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 50 μg/L (limit established for PAT in juices) and 25 μg/kg (limit established for PAT in purees), digested with an in vitro gastrointestinal protocol and bioaccessibility values (%) were calculated.     

Risk analysis of main mycotoxins occurring in food for children: An overview

Mycotoxins are secondary metabolites produced by fungi contaminating the food chain that are toxic to animals and humans. Children up to 12 years old are recognized as a potentially vulnerable subgroup with respect to consumption of these contaminants. Apart from having a higher exposure per kg body weight, they have a different physiology from that of adults. Therefore they may be more sensitive to neurotoxic, endocrine and immunological effects. For these reasons, a specific and up-to-date risk analysis for this category is of great interest.In this review, an accurate analysis of the main mycotoxins occurring in food intended for children (deoxynivalenol, aflatoxins, ochratoxins, patulin and fumonisins) is presented. In particular, known mechanisms of toxicity and levels of exposure and bioaccessibility in children are shown. In addition, recent discoveries about the strategies of mycotoxins managing are discussed.     

Genotypic identification of Penicillium expansum and the role of processing on patulin presence in juice

This work aimed at isolation and identification of patulin producing fungi and to follow the presence of patulin during apple juice processing. Among 34 Penicillium isolates, eight isolates (five from healthy appeared apples and 12 from rot spotted apples) were considered as patulin producers using thin-layer chromatography. These isolates were classically identified as a Penicillium expansum. PCR utilizing primers based on the polygalacturonase gene of P. expansum was applied for detecting this mold. The PCR amplified a 404-bp DNA product from all tested P. expansum isolates, but not in other common food spoilage Penicillium species. RAPD technique using P1 or M13 primers was applied to determine the similarity of the P. expansum isolates. RAPD results revealed that the tested strains showed high percentage of similarity and no correlation was observed between cluster analysis and the sources of isolation. Patulin could not be detected in healthy appeared apples and their extracted juice during different stages of juice process. In apple juice made from the healthy parts of apples decayed by P. expansum contained patulin which may present health hazard. The obtained results assured that patulin is known to be stable in apple juice even after pasteurization. In conclusion, the removal of the rotten part from the fruit is not sufficient to eliminate the mycotoxin patulin from apple juice. Although, the enzyme treatment (pectinase and amylase) and pasteurization (95 °C for 7 min) significantly (p < 0.05) reduced patulin level, its level is still higher than the level of <50 μg/kg considered by Codex alimentarius when the apple juice processed from the healthy parts of rot spotted fruits.     

Patulin causes DNA damage leading to cell cycle arrest and apoptosis through modulation of Bax, p and p proteins in skin of mice

Patulin (PAT), a mycotoxin found in apples, grapes, oranges, pear and peaches, is a potent genotoxic compound. WHO has highlighted the need for the study of cutaneous toxicity of PAT as manual labour is employed during pre and post harvest stages, thereby causing direct exposure to skin. In the present study cutaneous toxicity of PAT was evaluated following topical application to Swiss Albino mice. Dermal exposure of PAT, to mice for 4 h resulted in a dose (40-160 μg/animal) and time (up to 6 h) dependent enhancement of ornithine decarboxylase (ODC), a marker enzyme of cell proliferation. The ODC activity was found to be normal after 12 and 24 h treatment of patulin. Topical application of PAT (160 μg/100 μl acetone) for 24-72 h caused (a) DNA damage in skin cells showing significant increase (34-63%) in olive tail moment, a parameter of Comet assay (b) significant G 1 and S-phase arrest along with induction of apoptosis (2.8-10 folds) as shown by annexin V and PI staining assay through flow cytometer. Moreover PAT leads to over expression of p^21/WAF1 (3.6-3.9 fold), pro apoptotic protein Bax (1.3-2.6) and tumor suppressor wild type p⁵³ (2.8-3.9 fold) protein. It was also shown that PAT induced apoptosis was mediated through mitochondrial intrinsic pathway as revealed through the release of cytochrome C protein in cytosol leading to enhancement of caspase-3 activity in skin cells of mice. These results suggest that PAT has a potential to induce DNA damage leading to p⁵³ mediated cell cycle arrest along with intrinsic pathway mediated apoptosis that may also be correlated with enhanced polyamine production as evident by induction of ODC activity, which may have dermal toxicological implications.     

Patulin-induced genotoxicity and modulation of glutathione in HepG2 cells

Patulin (PAT), a mycotoxin produced by certain species of Penicillium, Aspergillus and Byssochlamys, is mainly found in ripe apple and apple products. In our present study, a significant increase of the micronuclei frequency induced by PAT was found in human hepatoma HepG2 cells. To elucidate the role of glutathione (GSH) in the effect, the intracellular GSH level was modulated by pre-treatment with buthionine-(S, R)-sulfoximine (BSO), a specific GSH synthesis inhibitor, and by pre-treatment with N-acetylcysteine (NAC), a GSH precursor. It was found that depletion of GSH in HepG2 cells with BSO dramatically increased the PAT-induced micronuclei frequencies and that when the intracellular GSH content was elevated by NAC, the chromosome damage induced by PAT was significantly prevented in our test concentrations (0.19-0.75 μM). These results indicate that GSH play an important role in cellular defense against PAT-induced genotoxicity.     

Tissue oxidative stress induced by patulin and protective effect of crocin

Patulin (PAT) is a secondary toxic metabolite produced principally by Penicillium expansum. This mycotoxin is known to be teratogenic, mutagenic, immunotoxic and neurotoxic, and it has been shown to cause damage in several organs in laboratory animals. This study focuses on the prevention of experimental murine PAT-induced nephrotoxicity and hepatotoxicity. We investigate the ability of a natural product, crocin (CRO), to counteract the toxic effects of PAT. Pre-treatment of mice with CRO prevented PAT-induced oxidative damage in both liver and kidney. CRO reduced lipid peroxidation, protein oxidation and restored redox status by regulating the endogenous antioxidant enzymatic system. These data corroborate and extend findings in PAT-induced nephrotoxicity and hepatotoxicity, and further suggest that preventive effect of CRO towards other forms of PAT toxicity, including neurotoxicity, may be warranted.     

高效液相色谱法测定果汁中展青霉素的含量

本文介绍了用高效液相色谱法测定展青霉素的含量，用C18反相柱，流动相乙腈 水(20 80)，流速1ml／min，紫外检测器，检测波长210nm。线性范围0～10ng(r=0．999)最低检出量为0．2ng，本方法定量准确、快速、主峰与杂质峰分离度高。     

展青霉素检测方法及污染情况的研究进展

本文对展青霉素近年来的检测方法及污染情况进行了综述,分析各种检测方法的优缺点.目前最常用的检测方法是高效液相色谱法,建立快速廉价的免疫学检测方法是今后的发展方向.另外展青霉素的污染情况不仅存在于苹果和山楂制品中,还存在于一些中药材(红枣、杏仁和木瓜等)中,多存在于霉烂水果、果汁当中.     

Crocin and quercetin prevent PAT‐induced apoptosis in mammalian cells: Involvement of ROS‐mediated ER stress pathway

Patulin (PAT) is a secondary metabolite produced by several species of the genera of Penicillium, Aspergillus, and Byssochlamys that can be found in rotting fruits, especially in apples and apple‐based products. Exposure to this mycotoxin has been reported to induce intestinal and kidney injuries. The mechanism underlying such toxicity has been linked to the induction of apoptosis which occurred with reactive oxygen species production and endoplasmic reticulum (ER) stress induction. This study aimed to evaluate the effect of the two common dietary compounds Quercetin (QUER), a natural flavonoid, and Crocin (CRO), a natural carotenoid, on PAT‐induced toxicity in human colon carcinoma (HCT116) and embryonic kidney cells (HEK293). We showed that antioxidant properties of QUER and CRO help to prevent ER stress activation and lipid peroxidation as evidenced by the reduction in GRP78 and GADD34 expressions and the decrease in malondialdehyde production. Furthermore, we demonstrated their ability to re‐establish the loss of the mitochondrial membrane potential to inhibit caspase 3 activation and DNA fragmentation. © 2015 Wiley Periodicals, Inc. Environ Toxicol 31: 1851–1858, 2016.