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BACKGROUND: In plastic surgery tissue transplantation, tissue ischemia limits transplanted tissue survival. Adipose-derived stem cells (ASCs) and stromal vascular fraction (SVF) show potential for promoting angiogenesis and rescuing ischemic conditions. However, when SVF and ASC suspensions are utilized without the protection of extracellular matrix, the retention rate of transplanted cells tends to be diminished, leading to an unsatisfactory therapeutic outcome. To overcome this, adipose tissue-derived microvascular fragments (ad-MVFs) have emerged as a promising solution. METHODS: We conducted enzymatic digestion on human adipose tissue to generate ad-MVFs. These fragments underwent a thorough characterization process, utilizing electron microscopy to assess their structural attributes and enabling a detailed analysis of their intricate morphology. Furthermore, our team investigated the cellular composition of these microvascular fragments, subsequently confirming their ability to enhance the viability of ischemic skin flaps. RESULTS: The resulting product primarily comprised fragments with sizes ranging from 20 to 50 µm, and some exhibited a sophisticated network-like structure. Electron microscopy examination revealed the presence of collagen components in the product. Additionally, flow cytometry analysis indicated a substantial abundance of adipose-derived stem cells and endothelial cells within these microvascular fragments. Significantly, when tested in treating an ischemic skin flap in a nude mouse model, the product exhibited superior therapeutic efficacy compared to SVF cell suspension. CONCLUSION: We have successfully generated human ad-MVFs and established standardized procedures. Compared with SVF, Ad-MVFs have a better effect in the treatment of ischemic diseases. LEVEL OF EVIDENCE II: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

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Quantifying manganese (Mn) content in solids is critical for understanding its roles in aquatic ecosystems, soils, water treatment plants and distribution systems. No studies have yet used standard Mn oxides to compare the performance of the numerous digestion methods found in the literature. Nine digestion methods (including USEPA 3050B) were compared using four Mn oxides with varying oxidation states. The HCl concentrate (12.4 M) heated to at least at 40 °C provided quantitative digestion of all Mn oxides tested with ≈ 100 % recovery. HCl concentration is important only for MnO2 digestion, while temperature influences both MnO and MnO2 recovery. Complete recovery of various Al, Cu and Fe standard oxides using a 12.4 M HCl digestion at 95 °C. Digestion of environmental samples for Al, Ca, Fe, Mg and Mn content yielded higher metal content using the HCl method (except for Al). HCl 12.4 M digestion provided better performance than other digestion methods found in the scientific literature because of its high reducing capacity. •Most digestion methods found in the literature do not digest all Mn oxidation states.•Hydrochloric acid is shown to be essential to dissolve all oxidation state of Mn oxides.

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To predict the apparent total tract digestibility (ATTD) of crude protein (CP) in dogs we developed an in vitro system using an in vitro digestion method and a statistical analysis. The experimental diets used chicken meat powder as the protein source, with CP levels of 20% (22.01%, analyzed CP value as dry-based), 30% (31.35%, analyzed CP value as dry-based), and 40% (41.34%, analyzed CP value as dry-based). To simulate in vivo digestive processes a static in vitro digestion was performed in two steps; stomach and small intestine. To analyze ATTD the total fecal samples were collected in eight neutered beagle dogs during the experimental period. CP digestibility was calculated by measuring CP levels in dog food, in vitro undigested fraction, and dog feces. In result, CP digestibility at both in vivo and in vitro was increased with increasing dietary CP levels. To estimate in vivo digestibility the co-relation of in vivo ATTD and in vitro digestibility was investigated statistically and a regression equation was developed to predict the CP ATTD (% = 2.5405 × in vitro CP digestibility (%) + 151.8). The regression equation was evaluated its feasibility by using a commercial diet. The predicted CP digestibility which was calculated by the regression equation showed high index of similarity (100.16%) with that of in vivo in dogs. With that, it would be a feasible non-animal method to predict in vivo CP digestibility by using in vitro digestion method and the proposed linear regression equation in adult dogs.

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A method for the in vitro isolation, purification, identification, and induced differentiation of satellite cells from adult tree shrew skeletal muscle was established. The mixed enzyme digestion method and differential adhesion method were used to obtain skeletal muscle satellite cells, which were identified and induced to differentiate to verify their pluripotency. The use of a mixture of collagenase II, hyaluronidase IV, and DNase I is an efficient method for isolating adult tree shrew skeletal muscle satellite cells. The P3 generation of cells had good morphology, rapid proliferation, high viability, and an "S"-shaped growth curve. Reverse transcription-polymerase chain reaction (RT-PCR) and immunofluorescence staining indicated that marker genes or proteins were expressed in skeletal muscle satellite cells. After myogenic differentiation was induced, multiple-nucleated myotubes were observed, and the MyHC protein was expressed. The expression of myogenic marker genes changed with the differentiation process. After the induction of adipogenic differentiation, orange-red lipid droplets were observed, and the expression of adipogenic marker genes increased gradually with the differentiation process. In summary, satellite cells from adult tree shrew skeletal muscle were successfully isolated using a mixed enzyme digestion method, and their potential for differentiation into myogenic and adipogenic cells was confirmed, laying a foundation for further in vitro study of tree shrew muscle damage.

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A novel protocol for the recovery of PHA from mixed-cultures proposed. In this experiment, activated sludge for PHA synthesis was investigated and a two-stage chemical digestion method was used for activated sludge to improve the yield of PHA. The highest PHA extraction combination that could be obtained in this experiment was sodium hypochlorite(NaClO) plus sodium dodecyl sulfate (SDS), and the optimal concentration of NaClO solution was 25 % (v/v), and the ratio of the dry weight of activated sludge to SDS was 1:2. The recovery and purity of PHA were 72.14 % and 54.47 %, respectively. The reaction time between NaClO and activated sludge affects the recovery of PHA, and the optimal reaction time of NaClO was experimentally obtained as 30 min. The purity of the PHA extract obtained after purification using methanol was improved.

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In total, two experiments were conducted to evaluate the effectiveness of an in vitro digestion method for predicting the metabolizable energy (ME) and metabolizability of gross energy (ME/GE) values using in vitro digestible energy (IVDE) and the digestibility of gross energy (IVDE/GE) content, respectively, of conventional feedstuffs for Muscovy ducks. In experiment 1, the apparent metabolizable energy (AME), true metabolizable energy (TME), AME/GE, and TME/GE of eight-grain feedstuff samples (two corn samples, three sorghum samples, and three barley samples) and eight protein feedstuff samples (two soybean meal samples, three cottonseed meal samples, and three rapeseed meal samples) were determined by the tube-feeding method with six different ducks for each sample. In experiment 2, a computer-controlled simulated digestion system (CCSDS) contain simulated digestive fluid was used to determine the enzymatic hydrolysis energy value of feedstuffs, which was defined as IVDE in our study. The simulated gastric fluid containing pepsin and simulated small intestinal fluid containing amylase, trypsin, and chymotrypsin for the in vitro gastric and intestinal digestion, respectively. The IVDE and in vitro digestibility of GE (IVDE/GE) of 16 feedstuff samples were determined using the CCSDS with five replicates per sample. The results showed that the IVDE and IVDE/GE were positively correlated with ME and ME/GE of feedstuffs, respectively. The coefficient of determination of eight regression models in predicting ME (grain feedstuffs: AME = 1.050 × IVDE- 0.9293, TME = 1.032 × IVDE + 0.6478; protein feedstuffs: AME = 1.331 × IVDE- 6.685, TME = 1.269 × IVDE-3.490) and ME/GE (grain feedstuffs: AME/GE = 1.069 × IVDE/GE- 6.516, TME/GE = 1.068 × IVDE/GE + 0.7764; protein feedstuffs: AME/GE = 1.093 × IVDE/GE -19.21, TME/GE = 1.196 × IVDE/GE - 13.25) of feedstuffs for Muscovy ducks ranged from 0.8610 to 0.9921. The accuracy of the regression model was acceptable as the difference between measured and predicted ME and ME/GE values was less than 0.45 MJ/kg (100 kcal/kg) and 2.62% for 14 of the 16 feed samples, respectively. In conclusion, the in vitro digestion method can be used to predict the ME and ME/GE of conventional feedstuffs for Muscovy ducks with acceptable accuracy.

Metabolizable energy (ME) is one of the major factors in formulating diets for ducks and most studies on the ME values of ingredients have been conducted on Peking ducks, with limited research on Muscovy ducks. Compared with the time-consuming in vivo digestion method, in vitro simulating digestion as a rapid and reliable method has been performed to predict ME and metabolizability of gross energy. Therefore, the precision of the in vitro digestion method was evaluated for Muscovy duck feed in our study.

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Microplastics (MPs) are a serious threat to the marine environment affecting ecosystem functioning and biodiversity. There is a vast literature about the uptake of MPs at different trophic levels, mainly focused on ecotoxicological effects in commercially relevant species. Little is still known about possible strategies to face MP pollution. Bioremediation is recently gaining attention in this framework. The clearance rate and particle retention of Sabella spallanzanii, Mytilus galloprovincialis, Phallusia mammillata, Paraleucilla magna at three MP concentrations (C1: 1.4 · 101 p/L; C2: 1.4 · 102 p/L; C3: 1.4 · 103 p/L) were investigated to test their potential as MP remover. Digestion protocol removed 98 % of tissues simplifying the MP quantification. P. magna clearance rate decreased with increasing concentration while P. mammillata showed no significant variations. S. spallanzanii and M. galloprovincialis instead exhibited the highest values of clearance rate. Yet, unlike mussels, S. spallanzanii can inhibit particle return to the surrounding water storing them in the tube, resulting to be the best candidate for bioremediation purposes.

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Plastic products are widely used in various fields, and the discarded plastics in the environment can be degraded into microplastics (MPs) or even nanoplastics (NPs), which significantly increases the risk of organism exposure. MPs/NPs have been found in aquatic organisms, birds of prey, and even humans. The detection of plastic particles in biological samples is more complicated than that in environmental samples. Biological samples are mainly composed of various organic substances such as proteins and lipids, which makes the pretreatment process particularly critical. Effective detection and accurate quantification of MPs/NPs are crucial to health risk assessment. In this paper, the exposure levels and composition of MPs/NPs in different tissues and organs of the human body, aquatic organisms, and birds of prey were reviewed. The digestion of biological samples (acids, bases, enzymes, and hydrogen peroxide digestion protocols) and MPs/NPs identification methods (spectroscopy and chromatography) were compared and their advantages and disadvantages were assessed, providing a methodological basis for plastic exposure risk assessment and pollution control.

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Trichinella spp. are foodborne parasites that can cause severe and potentially fatal disease in humans. Infections occur through consumption of meat containing the infectious stage (L1). In Germany the domestic cycle has been eradicated. In wild animals sporadic occurrence is observed in species such as wild boar, red foxes and raccoon dogs. The omnivore raccoon which is an invasive species in Europe is known as a potential host but has not been studied intensely regarding this parasite in Germany until now, thus resulting in a lack of knowledge about its role in the sylvatic cycle. Raccoons from the urban area of Leipzig were investigated for several pathogens including Trichinella spp. in a cooperative project. Muscle samples of 88 individuals were examined using the artificial digestion method (ADM). One animal was found positive, which is the first detection of this parasite in a raccoon in Germany.

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Trichinellosis is an important worldwide foodborne zoonosis. The gold standard test to detect Trichinella spp. larvae in muscle samples of animals intended for human consumption is the artificial digestion method. Handling and dispensing of conventional pepsin powder present significant safety risks for analysts. The use of pepsin powder that is resistant to aerosolization should alleviate these safety concerns. The aim of this study was to compare the efficacy of an aerosol-resistant pepsin powder to conventional pepsin powder in the artificial digestion method. Proficiency samples of pork diaphragm containing specific numbers of viable Trichinella spiralis larvae were tested in two laboratories. The results revealed that aerosol-resistant pepsin was simple, effective and convenient to use, and showed good solubility and larval recovery that met the requirements of the European Union regulation EU 2015/1375. Overall, the efficacy of the aerosol-resistant pepsin was comparable to the conventional pepsin and safer for analysts.

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Soy isoflavones, at adequate dosages, have estrogenic and anti-thyroidal effects in animals and humans, which can either be beneficial or adverse, depending on the consumer's physiological status. Hence, this study presents an assay of soy isoflavones in hair, aiming to give new information about a person's exposure to isoflavones, when health issues related to estrogenic or thyroidal effects are observed. Aqueous or organic extraction procedures following acidic, basic, or enzymatic digestions were tested on 60 hair samples (from volunteers) from a hairdresser, and a clinical trial 2017T2-29. The acidic digestion method was the most efficient regarding isoflavones. A specific inquiry was developed to assess the dietary habits of French consumers based on the analysis of 12,707 food labels from France. It was used to check for the reliability of the new assay method. A score for the consumer exposures to isoflavones was built considering, among other parameters, soy-based diets and foodstuff containing soy as an ingredient, i.e., "hidden-soy". The correlation between this score and isoflavone measurements in hair reached 0.947; p < 0.001. Therefore, providing that relevant data are considered to assess isoflavone exposure, hair that smoothens daily isoflavone intake variations, is a relevant tissue to assess human isoflavone exposure for subsequent health analyses.

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Complementary feeding starts at around six months of age because neither breast milk nor formula assure the proper nutrition of infants. Therefore, along with breast milk, solid foods are gradually introduced, particularly cereal-based foods, which will provide starch as a new source of energy and nutrients. As a result, the need of an adequate in vitro digestion method to study the influence of different aspects of weaning period is unquestionable. This critical review summarizes the in vitro digestion methods available for the analysis of starch hydrolysis under infant conditions considering different features, namely, starch digestion, infant digestive conditions and in vitro models suitable for the study of starch digestion (static, semi-dynamic and dynamic). Key factors such as enzyme concentrations, transit time, oral, gastric and intestinal conditions and differences with current adult models, have been addressed. The need for standardized infant digestion models adapted to the complementary feeding period was discussed. Existing literature data demonstrate that more effort has to be done to improve the research on this issue, in order to obtain comparable results that would address a better understanding of the digestibility of different food nutrients under infant conditions facilitating the development of appropriate formulations that may assure proper infant nutrition.

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Gluten proteins are the causative agents of celiac disease (CD), a lifelong and worldwide spread food intolerance, characterized by an autoimmune enteropathy. Gluten is a complex mixture of high homologous water-insoluble proteins, characterized by a high content of glutamine and proline amino acids that confers a marked resistance to degradation by gastrointestinal proteases. As a consequence of that, large peptides are released in the gut lumen with the potential to activate inflammatory T cells, in CD predisposed individuals. To date, several strategies aimed to detoxify gluten proteins or to develop immunomodulatory drugs to recover immune tolerance to gluten are under investigation. This review overviews the state of art of both analytical and functional methods currently used to assess the immunogenicity potential of gluten proteins from different cereal sources, including native raw seed flours and complex food products, as well as drug-treated samples. The analytical design to assess the content and profile of gluten immunogenic peptides, described herein, is based on the oral-gastro-intestinal digestion (INFOGEST model) followed by extensive characterization of residual gluten peptides by proteomic and immunochemical analyses. These approaches include liquid chromatography-high-resolution mass spectrometry (LC-MS/MS) and R5/G12 competitive ELISA. Functional studies to assess the immune stimulatory capabilities of digested gluten peptides are based on gut mucosa T cells or peripheral blood cells obtained from CD volunteers after a short oral gluten challenge.

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OBJECTIVE: To determine the efficacy of using sodium hypochlorite (NaClO, Purelox) as a simple and rapid alternative digestion method for the diatom test through a quantitative comparison with the conventional method that uses fuming nitric acid (HNO3). MATERIALS AND METHODS: In experiment 1, using 30 water samples, the NaClO and HNO3 methods were compared using paired t-test. In experiments 2 and 3, we included blank human lung plus 13 water samples and total of 32 drowned human lung samples, respectively, to compare the NaClO and HNO3 methods using paired t-test. The relationship between the concentration ratio and background factors was tested in experiment 3. Welch's t-test was used to determine differences in the ratio between the lung side and sex, whereas Pearson's correlation coefficient was used to determine the correlation between the ratio and either age or postmortem interval. The geometric mean of two counts was used for each specimen and all counts were logarithmically transformed to base 2 in the statistical analysis. RESULTS: The NaClO method was completed within 80 min for any sample. In experiment 1, there was no significant difference between the NaClO and HNO3 methods using water samples (the mean of the ratios: 0.99, 95% confidence interval (95%CI: 0.89-1.10, P = 0.80). In experiment 2, the count of the NaClO method was lower than that of the HNO3 method using lung plus water samples (the mean of the ratios: 0.47, 95% CI: 0.35-0.65, P = 0.0002). In experiment 3, the concentration of the NaClO method was lower than that of the HNO3 method using drowned lung samples (the mean of the ratios: 0.28, 95% CI: 0.20-0.38, P < 0.0001). A weak correlation between the postmortem interval and the ratio of the two methods was observed (r = -0.58, P = 0.012), although no difference between lung sides or sexes were detected (P = 0.87 and P = 0.50, respectively) and no correlation occurred between age and the ratio (r = 0.15, P = 0.43). CONCLUSION: Using NaClO as a simple and rapid digestion method for diatom testing of water samples would be an excellent alternative to conventional methods. Although the method's diatom detection rate for the lung samples was not optimal, it was still shown to be a feasible method.

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OBJECTIVES: The efficient acquisition and purification of fibroblasts as ideal seed cells are very important. For optimization of the isolation and culture of human foreskin fibroblasts (HFF), we compared the improved tissue culture method (ITCM) and the enzyme digestion method (EDM). METHODS: In ITCM, the skin tissue was digested with 0.1% Type II collagenase overnight at 4 ℃, the epidermis was separated from the dermis and digested again with 0.25% trypsin at room temperature for 15 min, and then the tissue block was attached to the culture dish. In EDM, the skin tissue was digested with 0.25% trypsin overnight at 4 ℃, the epidermis was separated from the dermis and digested with 0.1% Type II collagenase overnight at 4 ℃, the tissue block was filtered and squeezed together with the enzyme mixture, the filter was rinsed with medium containing fetal bovine serum, and the cell suspension was cultured. Both ITCM and EDM used 2 digestion enzymes, but the order, digestion time, and temperature of the 2 enzymes were different. The final inoculations of ITCM and EDM in the dishes for subsequent culture were tissue blocks and cell suspensions, respectively. In this study, HFF cells were isolated and cultured with ITCM and EDM, and the cell morphology was observed from Passage 0 to Passage 3 in the ITCM and EDM groups. The cell purity was identified by staining for vimentin, CD68, and Pan-keratin. The growth curves of Passage 3 were plotted to compare the proliferation ability of the 2 groups. Passage 3 HFF cells in the ITCM and EDM groups were irradiated with medium-wave ultraviolet (UVB) at an energy value of 120 mJ/cm2 to establish a light damage model. The experiments were grouped into an UVB group and a control group (Control) according to the presence or absence of UVB irradiation. Platelet-poor plasma (PPP) was extracted by secondary centrifugation, and the HFF cells of ITCM and EDM groups were cultured in groups using complete medium containing different concentrations (0, 2.5%, 5.0%, and 10.0%) of PPP, and the proliferation of damaged cells was detected by cell counting kit-8 after 24 h of PPP incubation. RESULTS: A large number of HFF could be observed in the ITCM group up to day 3, which was less affected by impurities; the observation of HFF morphology in the EDM group was affected by more impurities. By day 9, cells in both ITCM and EDM groups could be passaged; HFF isolated and cultured in vitro by the 2 methods showed long spindle-shaped, swirling growth. The positive rates of vimentin in the ITCM and EDM groups when HFF cells were cultured up to Passage 2 were significantly different [(97.36±0.76)% vs (99.4±0.56)%, P<0.01)]. The positive rates of CD68 were also significantly different [(70.8±0.46)% vs (78.37±0.75)%, P<0.01]. The expressions of pan-keratin in the ITCM group and the EDM group were positive and negative, respectively. There was no difference in vimentin and pan-keratin staining results between the ITCM group and the EDM group when HFF were cultured to Passage 3. The positive rates of CD68 between the ITCM group and the EDM group were significantly different [(74.73±1.37)% vs (85.27±2.63)%, P<0.001]. The proliferative capacity of HFF cells in Passage 3 was significantly higher in the EDM group than that in the ITCM group (P<0.05). After UVB (120 mJ/cm2) irradiation, HFFs procured by the 2 isolation methods showed damage. The damage repair test demonstrated that the 2.5% PPP+UVB irradiation group showed significantly higher repair competence than the other groups (all P<0.05). CONCLUSIONS: In contrast with HFFs isolated via ITCM, HFF cells isolated by EDM have a faster purification rate and a stronger proliferative capacity. Therapy with PPP can moderately repair UVB-induced damage to HFFs. The results provide a theoretical basis for clinical treatment studies in the future.

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This work studied the performance of the artificial digestion method in terms of recovery and viability of Anisakis simplex third-stage larvae (L3) when previous treatments given to the infected fish muscle may accidentally render viable larvae. For that: a) hake mince was spiked with 10 L3/75g mince, frozen at -10, -15, -20, and -30 °C and immediately thawed, or stored for 12 or 24 h, and subjected to pepsin digestion; b) the mince was spiked under the same conditions, frozen at the above temperatures and thawed immediately. After manual recovery, L3 were assessed for viability, used to spike again the minced fish and subjected to pepsin digestion; c) the mince was spiked with 10 L3 which were: i) living (i.e. chilled), ii) freeze-surviving (live L3 had been previously recovered after freezing at -10 °C), or iii) dead (frozen at -30 °C or - 80 °C), and then subjected to pepsin digestion. Results showed that the artificial digestion method kills a significant number of larvae that may have survived freezing and thus may underestimate the number of viable larvae in a given batch. The method may also underestimate the infection level of fish batches containing dead larvae. It is suggested to take these limitations into account when designing digestion protocols for specific applications, especially when there is a risk of insufficiently treated or cooked fish batches or ready-to-eat foods.

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Microplastics (MPs) are an emerging threat to marine ecosystems. One of the primary environmental risks is their bioavailability for aquatic organisms. Some fish and bivalves are of particular interest because their feeding strategies expose them to particles present in the water column. The aim of the study was to assess an extraction method in order to isolate and quantify MPs from fish gastrointestinal tract (n.8) and muscle (n.4), and bivalves (n.8) samples. The accuracy of the method was assessed through the calculation of the recovery percentage in samples spiked with a known number of MPs using microscopic observation. Successively, the extraction was preliminarily applied on n.20 mussels collected from mariculture plants of the Tyrrhenian and the Adriatic Sea. The results of the digestion protocol showed an average extraction yield of 80% in fish gastrointestinal tracts, 90% in fish muscle samples, and 95% in mussels. Preliminary analysis carried out on farmed mussels showed an average abundance of 3.8 items/individual, and 0.5 items/g of tissue, among those black, was the most represented color.

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Starch, dextran, pectin and modified citrus pectin were subjected to intestinal digestion following InfoGest protocol and a rat small intestine extract (RSIE) treatment. Gastric stage did not show any modification in the structure of the carbohydrates, except for modified pectin. Regarding intestinal phases, starch was hydrolyzed by different ways, resulting in a complementary behavior between InfoGest and RSIE. Contrarily, digestion of dextran was only observed using RSIE. Similar situation occurred in the case of pectins with RSIE, obtaining a partial hydrolysis, especially in the modified citrus pectin. However, citrus pectin was the less prone to hydrolysis by enzymes. The results demonstrated that InfoGest method underestimates the significance of the carbohydrates hydrolysis at the small intestine, thus indicating that RSIE is a very reliable and useful method for a more realistic study of polysaccharides digestion.

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OBJECTIVE：To establish the method for the content determination of inorganic elements in Cyperus rotundus ，and to compare the contents of 14 kinds of inorganic elements in C. rotundus from different producing areas ，and to provide theoretical basis for its quality control and high quality resources development . METHODS ：The samples were processed by microwave digestion，and ICP-MS method was used to determine the contents of Na ，Mg，K，Ca，Mn，Fe，Ni，Cu，Zn，As，Se，Sr，Cd and Pb. SPSS 23.0 software were used for principal component analysis （PCA）and cluster analysis. RESULTS ：The average contents of above 14 kinds of inorganic elements in C. rotundus from different producing areas were 168.62，753.71， 6 938.33，24.31，14.69，197.77，0.60，2.43，26.89，0.21，0.06，5.64，0.05，0.32 mg/kg，respectively. The results of PCA showed that the cumulative variance contribution rate of the first four principal components was 86.203％，which could reflect most of the information of the original data. C. rotundus from Shandong ，Jiangxi，Shanxi，Hubei and Yunnan ranked the top five places in terms of comprehensive score of inorganic element contents. The results of cluster analysis showed that the samples from 9 producing areas were clustered into 5 categories，showing the characteristics of clustering by producing area. From the perspective of inorganic elements ，the quality of C. rotundus from East China ，Central China ，North China and Southwest China was better than that from South China. CONCLUSIONS ：Essential trace elements like Na ，Mg，K，Ca，Mn，Fe，Cu，Zn，Sr are rich in C. rotundus，and there are small amounts of Ni ，As，Se，Cd，Pb elements in it. The contents difference of inorganic elements in C. rotundus from different origins may related to the geographical area it belongs to.

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Food contaminants are usually studied as isolated molecules, not considering the overall meal components. Notwithstanding, contaminants are not ingested individually, therefore their risks should be assessed in the context of the overall diet. In the present study the influence of three well known dietary patterns, Western (W), Mediterranean (M) and vegetarian (V), on the bioaccessibility and intestinal transport of polycyclic aromatic hydrocarbons (PAHs) (benzo[a]pyrene (BaP) and benzo[b]fluoranthene (BbF)), heterocyclic aromatic amines (HAAs) (2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) and 2-Amino-9H-pyrido[2,3-b]indole (AAC)) and mycotoxins (MY) (aflatoxin B2 (AB2) and ochratoxin A (OTA)) was evaluated. Whole meals representative of W, M and V patterns were spiked with 100 µg kg-1 of each contaminant and subjected to the Infogest in vitro digestion method. Intestinal transport was performed using Caco-2 cells in apical/basolateral inserts. Contaminants were quantified by QuEChERS/HPLC/Fluorescence analysis. The dietary pattern itself influenced significantly the bioaccessibility of some contaminants, since higher bioaccessibility of HAAs (PhIP and AAC) was observed for V diet, while higher bioaccessibility of PAHs (BBF and BAP) and the MY (OTA) was observed for W diet. Concerning intestinal transport, the effect of the diet matrices was less noticed. Notwithstanding, AAC transport increased with W diet, while AB2s transport increased with the V diet. Regarding PAHs the three patterns either blocked (BbF) or reduced (BaP) the transport. Besides the well known nutritional, protective or deleterious effects of the different dietary patterns, the increased bioaccessibility or intestinal transport of some food contaminants, can have an additional influence on the global health impact.

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