Identification of Novel Compounds Inhibiting the Kinase Activity of the CDK5/p25 Complex via Direct Binding to p25.

Tamoxifen, the gold standard drug for endocrine therapy for breast cancer, modulates the phosphorylation status of the TAU protein in Alzheimer's disease by inhibiting CDK5 kinase activity. Its binding to p25 prevents CDK5/p25 complexation and hence a decrease of CDK5 activity. In breast tumors, this complex is involved in the proliferation and survival of cancer cells, as well as in the disease's prognosis. Still, the molecular stability of the CDK5/p25 complex following tamoxifen exposure in this cancer type has not yet been clearly deciphered. Here, we report the functional characterization of CDK5 and its p25 regulatory subunit in the absence and presence of tamoxifen. In addition, two novel inhibitors of the kinase activity of the CDK5/p25 complex are identified, both of which would reduce the risk of recurrence of estrogen receptor-positive (ER+) breast cancers and prevent drawbacks induced by tamoxifen exposure. Accordingly, 6His-CDK5 and 6His-p25 have been expressed and purified. Fluorescence anisotropy measurements have been used to assess that the two proteins do form an active complex, and thermodynamic parameters of their interaction were measured. It was also confirmed that tamoxifen directly binds to p25 and inhibits CDK5 kinase activity. Similar observations were obtained using 4-hydroxytamoxifen, an active metabolized form of tamoxifen. Two novel compounds have been identified here that harbor a benzofuran moiety and were shown to target directly p25, and their bindings resulted in decreased CDK5 kinase activity. This encouraging alternative opens the way to the ensuing chemical optimization of this scaffold. It also promises a more specific therapeutic approach that may both tackle the pathological signaling in breast cancer and provide a potential new drug for Alzheimer's disease.

Curcumin and NCLX inhibitors share anti-tumoral mechanisms in microsatellite-instability-driven colorectal cancer.

BACKGROUND AND AIMS: Recent evidences highlight a role of the mitochondria calcium homeostasis in the development of colorectal cancer (CRC). To overcome treatment resistance, we aimed to evaluate the role of the mitochondrial sodium-calcium-lithium exchanger (NCLX) and its targeting in CRC. We also identified curcumin as a new inhibitor of NCLX. METHODS: We examined whether curcumin and pharmacological compounds induced the inhibition of NCLX-mediated mitochondrial calcium (mtCa2+) extrusion, the role of redox metabolism in this process. We evaluated their anti-tumorigenic activity in vitro and in a xenograft mouse model. We analyzed NCLX expression and associations with survival in The Cancer Genome Atlas (TCGA) dataset and in tissue microarrays from 381 patients with microsatellite instability (MSI)-driven CRC. RESULTS: In vitro, curcumin exerted strong anti-tumoral activity through its action on NCLX with mtCa2+ and reactive oxygen species overload associated with a mitochondrial membrane depolarization, leading to reduced ATP production and apoptosis. NCLX inhibition with pharmacological and molecular approaches reproduced the effects of curcumin. NCLX inhibitors decreased CRC tumor growth in vivo. Both transcriptomic analysis of TCGA dataset and immunohistochemical analysis of tissue microarrays demonstrated that higher NCLX expression was associated with MSI status, and for the first time, NCLX expression was significantly associated with recurrence-free survival. CONCLUSIONS: Our findings highlight a novel anti-tumoral mechanism of curcumin through its action on NCLX and mitochondria calcium overload that could benefit for therapeutic schedule of patients with MSI CRC.

Estrogenic hazards of short chain phthalates and bisphenols found in cosmetic products.

For several years, environmental exogenous agents, called endocrine disruptors, are suspected to interfere with the essential functions of reproduction and development in many living organisms. In this study, endocrine disruptors including five phthalates and two bisphenols contents in finished products were assayed and their estrogenic activity were measured by using the Yeast Estrogen Screen system with respect to human and trout estrogen receptors hERα and rtERS. Independently of the estrogen receptor, only short-chain phthalates (DBP and BBP) and the two bisphenols exhibited an estrogenic activity. Besides, the risk of three end-products (agro-food, cosmetics, and pharmaceutical) was evaluated before and after forced aging. Only two cosmetics the face cream and the perfume presented a hazard which increases with aging. These results are consistent with the compounds identified by Gas chromatography-mass spectrometry. These findings confirmed that the YES system can be routinely used to evaluate the estrogenic hazards within finished products.

Ruling out septic arthritis risk in a few minutes using mid-infrared spectroscopy in synovial fluids.

OBJECTIVES: The aim of this study was to show the usefulness of a mid-infrared fibre evanescent wave spectroscopy point of care device in the identification of septic arthritis patients in a multicentre cohort, and to apply this technology to clinical practice among physicians. METHODS: SF samples from 402 patients enrolled in a multicentre cohort were frozen for analysis by mid-infrared fibre evanescent wave spectroscopy. The calibration cohort was divided into two groups of patients (septic arthritis and non-septic arthritis) and relevant spectral variables were used for logistic regression model. Model performances were tested on an independent set of 86 freshly obtained SF samples from patients enrolled in a single-centre acute arthritis cohort and spectroscopic analyses performed at the patient's bedside. RESULTS: The model set-up, using frozen-thawed SFs, provided good performances, with area under the curve 0.95, sensitivity 0.90, specificity 0.90, positive predictive value 0.41 and negative predictive value 0.99. Performances obtained in the validation cohort were area under the curve 0.90, sensitivity 0.92, specificity 0.81, positive predictive value 0.46 and negative predictive value 0.98. The septic arthritis probability has been translated into a risk score from 0 to 4 according to septic risk. For a risk score of 0, the probability of identifying a septic patient is very low (negative predictive value of 1), whereas a risk score of 4 indicates very high risk of septic arthritis (positive predictive value of 1). CONCLUSION: Mid-infrared fibre evanescent wave spectroscopy could distinguish septic from non-septic synovial arthritis fluids with good performances, and showed particular usefulness in ruling out septic arthritis. Our data supports the possibility of technology transfer. TRIAL REGISTRATION: ClinicalTrials.gov, http://clinicaltrials.gov, NCT02860871.

Development of a new approach using mathematical modeling to predict cocktail effects of micropollutants of diverse origins.

A wide variety of micropollutants (MP) of diverse origins is present in waste and surface waters without knowing the effect of their combination on ecosystems and human. The impact of chemical mixtures is poorly documented and often limited to binary mixtures using MP of the same category. Knowing that it is not realistic to test every possible combination found in mixtures, we aimed to develop a new method helping to predict cocktail effects. Six chemicals of agriculture, industry or pharmaceutical origin were selected: cyproconazole, diuron, terbutryn, bisphenol A, diclofenac and tramadol. Individual MP were first used in vitro to determine the concentration at which 10% (Effective Concentration EC10) or 25% (EC25) of their maximal effect on human cytotoxicity was observed. Using an Orthogonal Array Composite Design (OACD), relevant complex mixtures were then tested. Multiple linear regression was applied for response surface modeling in order to evaluate and visualize the influence of the different MP in mixtures and their potential interactions. The comparison of the predicted values obtained using the response surface model with those obtained with the model of independent effects, evidenced that the hypothesis of independence was unjustified. The cocktail effect was further investigated by considering micropollutant response surfaces pairwise. It was deduced that there was a neutralizing effect between bisphenol A and tramadol. In conclusion, we propose a new method to predict within a complex mixture of MP the combinations likely involved in cocktail effects. The proposed methodology coupling experimental data acquisition and mathematical modeling can be applied to all kind of relevant bioassays using lower concentrations of MP. Situations at high ecological risk and potentially hazardous for humans will then be identified, which will allow to improve legislation and policies.

Designation of pathogenic resistant bacteria in the Sparusaurata sea collected in Tunisia coastlines: Correlation with high performance liquid chromatography-tandem mass spectrometry analysis of antibiotics.

Vibrio is characterized by a large number of species and some of them are human pathogens causing gastro intestinal and wound infections through the ingestion or manipulation of contaminated fishes including Vibrio parahaemolyticus and Vibrio alginolyticus. In this study, we reported the phenotypic and molecular characterization of Vibrio parahaemolyticus and Vibrio alginolyticus strains isolated from wild and farm sea bream (Sparus aurata L.) along the Tunisian coast from December 2015 to April 2016. Therefore, the antibiograms indicate a difference between farmed and wild fish. Resistance against amoxicillin antibiotic appears for the bacteria isolated from wild fish, while those from aquaculture farming presented sensitivity to amoxicillin and resistance to antibiotics colistin and fusidic acid. The chloramphenicol antibiotic exhibited a high sensitivity in all isolated bacteria. In fact, traces of amoxicillin in the organs of the fish from Hergla farm were detected by UPLC-MS/MS analysis during December 2016 to April 2016. In addition, antibiotics were detected in January 2014 with high concentration of norfloxacin 2262 ng/g in fish from Hergla coast. The results obtained in this work indicated that the use and presence of antibiotics in water impacts on the occurrence of resistant bacteria and the detection of antibiotic in fish.

Mid-infrared fibre evanescent wave spectroscopy of serum allows fingerprinting of the hepatic metabolic status in mice.

Non-alcoholic fatty liver disease is associated with obesity, diabetes, and metabolic syndrome. The detection of systemic metabolic changes associated with alterations in the liver status during non-alcoholic fatty liver disease could improve patient follow-up. The aim of the present study was to evaluate the potential of mid-infrared fibre evanescent wave spectroscopy as a minimum-invasive method for evaluating the liver status during non-alcoholic fatty liver disease. Seventy-five mice were subjected to a control, high-fat or high-fat-high carbohydrate diets. We analysed the serum biochemical parameters and mRNA levels of hepatic genes by quantitative RT-PCR. Steatosis was quantified by image analysis. The mid-infrared spectra were acquired from serum, and then analysed to develop a predictive model of the steatosis level. Animals subjected to enriched diets were obese. Hepatic steatosis was found in all animals. The relationship between the spectroscopy-predicted and observed levels of steatosis, expressed as percentages of the liver biopsy area, was not linear. A transition around 10% steatosis was observed, leading us to consider two distinct predictive models (<10% and >10%) based on two different sets of discriminative spectral variables. The model performance was evaluated using random cross-validation (10%). The hypothesis that additional metabolic changes occur beyond this transition was supported by the fact that it was associated with increased serum ALT levels, and Col1α1 chain mRNA levels. Our data suggest that mid-infrared spectroscopy combined with statistical analysis allows identifying serum mid-infrared signatures that reflect the liver status during non-alcoholic fatty liver disease.

Oyster's cells regulatory volume decrease: A new tool for evaluating the toxicity of low concentration hydrocarbons in marine waters.

Human activities require fossil fuels for transport and energy, a substantial part of which can accidentally or voluntarily (oil spillage) flow to the marine environment and cause adverse effects in human and ecosystems' health. This experiment was designed to estimate the suitability of an original cellular biomarker to early quantify the biological risk associated to hydrocarbons pollutants in seawater. Oocytes and hepatopancreas cells, isolated from oyster (Crassostrea gigas), were tested for their capacity to regulate their volume following a hypo-osmotic challenge. Cell volumes were estimated from cell images recorded at regular time intervals during a 90min-period. When exposed to diluted seawater (osmolalities from 895 to 712mosmkg(-1)), both cell types first swell and then undergo a shrinkage known as Regulatory Volume Decrease (RVD). This process is inversely proportional to the magnitude of the osmotic shock and is best fitted using a first-order exponential decay model. The Recovered Volume Factor (RVF) calculated from this model appears to be an accurate tool to compare cells responses. As shown by an about 50% decrease in RVF, the RVD process was significantly inhibited in cells sampled from oysters previously exposed to a low concentration of diesel oil (8.4mgL(-1) during 24h). This toxic effect was interpreted as a decreased permeability of the cell membranes resulting from an alteration of their lipidic structure by diesel oil compounds. In contrast, the previous contact of oysters with diesel did not induce any rise in the gills glutathione S-transferase specific activity. Therefore, this work demonstrates that the study of the RVD process of cells selected from sentinel animal species could be an alternative bioassay for the monitoring of hydrocarbons and probably, of various chemicals in the environment liable to alter the cellular regulations. Especially, given the high sensitivity of this biomarker compared with a proven one, it could become a relevant and accurate tool to estimate the biological hazards of micropollutants in the water.

Dynamic extreme values modeling and monitoring by means of sea shores water quality biomarkers and valvometry.

Water quality can be evaluated using biomarkers such as tissular enzymatic activities of endemic species. Measurement of molluscs bivalves activity at high frequency (e.g., valvometry) during a long time period is another way to record the animal behavior and to evaluate perturbations of the water quality in real time. As the pollution affects the activity of oysters, we consider the valves opening and closing velocities to monitor the water quality assessment. We propose to model the huge volume of velocity data collected in the framework of valvometry using a new nonparametric extreme values statistical model. The objective is to estimate the tail probabilities and the extreme quantiles of the distribution of valve closing velocity. The tail of the distribution function of valve closing velocity is modeled by a Pareto distribution with parameter 𝜃 t,τ , beyond a threshold τ according to the time t of the experiment. Our modeling approach reveals the dependence between the specific activity of two enzymatic biomarkers (Glutathione-S-transferase and acetylcholinesterase) and the continuous recording of oyster valve velocity, proving the suitability of this tool for water quality assessment. Thus, valvometry allows in real-time in situ analysis of the bivalves behavior and appears as an effective early warning tool in ecological risk assessment and marine environment monitoring.

Investigating the in Vitro Thermal Stability and Conformational Flexibility of Estrogen Receptors as Potential Key Factors of Their in Vivo Activity.

Among hormone-inducible transcription factors, estrogen receptors (ERs) play important roles in tissue growth and differentiation, via either direct or indirect binding, in the nucleus, to specific DNA targets called estrogen responsive elements (EREs), or through nongenomic pathways. In humans, two estrogen receptor isoforms (hERs), designated hERα and hERß, have been identified. These two hERs, encoded by genes located on distinct chromosomes, exhibit divergent tissue-specific functions and different subcellular distributions depending on their binding status, free or complexed to their cognate ligands. Because it is hypothesized that such distinct behaviors may arise from various conformational stabilities and flexibilities, the effect of salt concentration and temperature was studied on the free and estrogen-activated hERα and hERß. Our results show that the conformational stability of hERß is weakly modulated by salt concentration as opposed to hERα. In addition, we show that the estrogen-bound hERs exhibit a more constrained structure than the unliganded ones and that their conformational flexibility is more affected by diethylstilbestrol binding than that of estradiol, 4-hydroxytamoxifen, or raloxifen. In line with these results, conformational analysis and computational docking were performed on hERα and hERß, which confer molecular support of a diethylstilbestrol-induced restrained flexibility as compared to other ligands. We found that Trp383 in hERα and Trp335 in hERß can closely interact with the NR-box motif of the H12 helix and act as a gatekeeper of the agonist-bound versus antagonist-bound conformations. Altogether, our study contributes to an improved knowledge of the diverse physicochemical properties of full-length hERs, which will help in our understanding of their distinct cellular roles in various cellular contexts.

Effect of a new regeneration process by adsorption-coagulation and flocculation on the physicochemical properties and the detergent efficiency of regenerated cleaning solutions.

Reprocessing soiled cleaning-in-place (CIP) solutions has large economic and environmental costs, and it would be cheaper and greener to recycle them. In food industries, recycling of CIP solutions requires a suitable green process engineered to take into account the extreme physicochemical conditions of cleaning while not altering the process efficiency. To this end, an innovative treatment process combining adsorption-coagulation with flocculation was tested on multiple recycling of acid and basic cleaning solutions. In-depth analysis of time-course evolutions was carried out in the physicochemical properties (concentration, surface tension, viscosity, COD, total nitrogen) of these solutions over the course of successive regenerations. Cleaning and disinfection efficiencies were assessed based on both microbiological analyses and organic matter detachment and solubilization from fouled stainless steel surfaces. Microbiological analyses using a resistant bacterial strain (Bacillus subtilis spores) highlighted that solutions regenerated up to 20 times maintained the same bactericidal efficiency as de novo NaOH solutions. The cleanability of stainless steel surfaces showed that regenerated solutions allow better surface wettability, which goes to explain the improved detachment and solubilization found on different types of organic and inorganic fouling.

A multi-scale approach of the mechanisms underlying exopolysaccharide auto-organization in the Proteus mirabilis extracellular matrix.

For decades, the origin of the concentric ring pattern of bacterial swarming colonies has puzzled microbiologists. It was hypothesized that a periodic water activity variation originates a phase transition within the extracellular matrix water H bond network, which switches on and off the exopolysaccharide auto-organization. Both rheological and infrared spectroscopy measurements respectively performed at a molecular scale and on a currently migrating colony, have given a physical insight into the mechanisms which underlie the switch between swarming and consolidation phases. Thanks to in situ and real time infrared microspectroscopy, and thanks to the brilliance of the infrared beam at SOLEIL synchrotron, here we demonstrate that Proteus mirabilis swarming is triggered by a periodic variation of water activity at the colony edge. A dynamic behavior emerges from the global properties of the multicellular entity which relies on the ability of the bacterial cells to tune exoproduct synthesis in order to undergo sharp transitions at a given water activity threshold.

Phenol-soluble modulins form amyloids in contact with multiple surface chemistries.

Functional amyloids are commonly produced by many microorganisms and their biological functions are numerous. Staphylococcus aureus can secrete a group of peptides named phenol-soluble modulins (PSMs) in their biofilm extracellular matrix. PSMs have been found inside biofilms both in their soluble form and assembled into amyloid structures. Yet, the actual biological function of these amyloids has been highly debated. Here, we assessed the ability of PSMs to form amyloids in contact with different abiotic surfaces to unravel a potential unknown bioadhesive and/or biofilm stabilization function. We combined surface plasmon resonance imaging, fluorescence aggregation kinetics, and FTIR spectroscopy in order to evaluate the PSM adsorption as well as amyloid formation properties in the presence of various surface chemistries. Overall, PSMs adsorb even on low-binding surfaces, making them highly adaptable adsorbants in the context of bioadhesion. Moreover, the PSM aggregation potential to form amyloid aggregates is not impacted by the presence of the surface chemistries tested. This versatility regarding adsorption and amyloid formation may imply a possible role of PSMs in biofilm adhesion and/or structure integrity.

Mid-Infrared Spectroscopy as a New Tool for Ruling Out Spontaneous Bacterial Peritonitis: A Proof-of-Concept Study.

BACKGROUND AND AIMS: A highly sensitive and specific point-of-care method for diagnosing spontaneous bacterial peritonitis (SBP) is currently lacking. The objective of the present study is to evaluate the diagnostic value of a rapid, easy-to-use, mid-infrared fiber evanescent wave spectroscopy (MIR-FEWS) method for ruling out SBP. PATIENTS AND METHODS: Cirrhotic patients (n = 256) at five centers in France were included for suspected SBP or for the scheduled evacuation of ascites fluid. The mid-infrared spectrum of 7 µL of an ascites fluid sample was recorded using a MIR-FEWS system. To define a model for the diagnosis of SBP, the patients were divided into a calibration group (n = 170) and a validation group (n = 86). RESULTS: Most of the patients were male (71%). The mean age was 60.25 years. Alcohol-related liver disease was the most common cause of cirrhosis. SBP was observed in 18% of the patients. For the diagnosis of SBP in the calibration and validation groups, respectively, the model gave areas under the receiver operating characteristic curves of 0.87 and 0.89, sensitivities of 90% and 87%, specificities of 78% and 80%, positive predictive values of 48% and 50%, negative predictive values of 97% and 96%, positive likelihood ratio of 4.09 and 4.35, negative likelihood ratio of 0.13 and 0.16, Youden index of 0.68 and 0.67, and correct classification rates of 80% and 81%. CONCLUSION: The results of this proof-of-concept study show that MIR-FEWS is a highly sensitive diagnostic method for ruling out SBP. The method warrants further investigation.

Control of vitellogenin genes expression by sequences derived from transposable elements in rainbow trout.

In most of oviparous animals, vitellogenins (VTG) are the major egg yolk precursors. They are produced in the liver under the control of estrogens. In rainbow trout (Oncorhynchus mykiss), the vtg genes cluster contains an unusually large number of almost identical gene copies. In order to identify the regulatory elements in their promoters, we used a combination of reporter plasmids containing genomic sequences including putative estrogen response elements (EREs) and we performed transient transfection assays in MCF-7 and yeast cells. We found a functional ERE corresponding to the sequence GGGGCAnnnTAACCT (rtvtgERE), which differs from the consensus ERE (ERE(cs)) by three base pairs. This non-palindromic ERE is located in the env gene of a retrotransposon relic, 180 base pairs upstream of the transcriptional start site. Fluorescence anisotropy experiments confirmed that the purified human estrogen receptor alpha (hERalpha) can specifically bind to rtvtgERE. Furthermore, we observe that the stability of hERalpha-ERE(cs) and hERalpha-rtvtgERE complexes is similar with equilibrium dissociation constants of 3.0nM and 6.2nM respectively, under our experimental conditions. Additionally, this rtvtgERE sequence displays a high E2-responsiveness through ER activation in cellulo. In the rainbow trout, the functional ERE (rtvtgERE) lies within promoter sequences which are mostly composed of sequences derived from transposable elements (TEs), which therefore may have acted as an evolutionary buffer to secure the proper expression of these genes.

An explanatory model to validate the way water activity rules periodic terrace generation in Proteus mirabilis swarm.

This paper explains the biophysical principles which, according to us, govern the Proteus mirabilis swarm phenomenon. Then, this explanation is translated into a mathematical model, essentially based on partial differential equations. This model is then implemented using numerical methods of the finite volume type in order to make simulations. The simulations show most of the characteristics which are observed in situ and in particular the terrace generation.

The mid-infrared spectroscopy: A novel non-invasive diagnostic tool for NASH diagnosis in severe obesity.

There is an urgent medical need to develop non-invasive tests for non-alcoholic steatohepatitis (NASH). This study evaluates the diagnostic performance of an innovative model based on mid-infrared (MIR) spectroscopy for the diagnosis of NASH. METHODS: Severely obese patients who underwent a bariatric procedure at the University Hospital of Nice, France (n = 395) were prospectively recruited. The clinico-biological characteristics were measured prior to surgery. Liver biopsies were collected during the surgical procedure and assessed by a pathologist. A training group (316 patients, NASH: 16.8%) and a validation group (79 patients, NASH: 16.5%) were randomly defined. MIR spectra were acquired by fiber evanescent wave spectroscopy, using chalcogenide glass fiber optic sensors and a spectrometer. This absorption spectroscopic technique delivers a spectrum that identifies the molecular composition of a sample, defining a patient's metabolic fingerprint. RESULTS: The areas under the receiver operating curve (AUROC) for the diagnosis of NASH were 0.82 and 0.77 in the training and validation groups, respectively. The best threshold was 0.15, which was associated with a sensitivity of 0.75 and 0.69, and a specificity of 0.72 and 0.76. Negative predictive values of 0.94 and 0.93 and positive predictive values of 0.35 and 0.36, as well as correctly classified patient rates of 72% and 75% were obtained in the training and validation groups, respectively. A composite model using aspartate aminotransferase level, triglyceride level and waist circumference alongside the MIR spectra led to an increase in AUROC (0.88 and 0.84 for the training and validations groups, respectively). CONCLUSIONS: MIR spectroscopy provides good sensitivity and negative predictive values for NASH screening in patients with severe obesity. LAY SUMMARY: There is an urgent need for tools to non-invasively diagnose and monitor non-alcoholic steatohepatitis (NASH). This study evaluates the performance of a new tool for fast NASH diagnosis based on mid-infrared (MIR) spectroscopy. Using serum samples from severely obese patients who underwent a bariatric procedure, which enabled a concomitant liver biopsy to be performed, the MIR spectroscopy model performed well in screening patients for NASH compared to a traditional, histological diagnosis.

Microplastics elutriation system: Part B: Insight of the next generation.

Elutriation is an efficient process for extracting microplastics. The development of a numerical model has shown the need for optimizing aspects of the design of the actual elutriation protocol as well as the dimensioning of the column to increase its efficiency. The study aims to propose new dimensioning data and protocol elements for designing an efficient column. Using a numerical model, the filling velocity was calculated as a function of the size and the density of the particles to prevent sand suspension. The sieving protocol was adapted to increase the density limit for the extraction of plastic particles from 1460 to >1800 kg·m-3. The durations of the elutriation and the column height were calculated to improve the control of the particle suspension. These results contribute to the development of the next generation of elutriation system and will accelerate the study of plasticome in the context of sandy sediments.

Persistent plasticizers and bisphenol in the cheese of Tunisian markets induced biochemical and histopathological alterations in male BALB/c mice.

Clinical evidences, experimental models, and epidemiology of many studies suggest that phthalate-based plasticizers, aliphatic ester, and bisphenol A (BPA) have major risks for humans by targeting different organs and body systems. The current study has been designed firstly to analyze three categories of cheese with and without their exposure to the sun and packed in packages with an inner surface plastic-covered film in order to identify the dibutyl phthalate (DBP); benzyl butyl phthalate (BBP); bis(2-ethylhexyl) phthalate (DEHP); diisodecyl phthalate (DiDP); diisononyl phthalate (DiNP); and 1,2-cyclohexane dicarboxylic acid diisononyl ester (DINCH) by GC-MS/MS, except for the bisphenol A, which is by UPLC-MS/MS, and secondly to assess the toxicity of the identified chemical molecules and cheese samples on the liver and kidney of mice. Our results showed that the cheese contains high quantities of DBP and DEHP with the concentrations up to 0.46 and 2.339 mg/kg, respectively. Other types of cheese, such as rolled and triangular cheeses, contain little quantities of the all substances at concentrations below the standard limits. In vivo, the obtained data clearly demonstrated that the acute administration of DBP, DEHP, and the tested cheese significantly induced liver and kidney injuries in mice manifested by a rise in plasma alanine aminotransferase, aspartate aminotransferase, blood urea nitrogen, lactate dehydrogenase, urea, creatinine, and uric acid when compared with control animals. In addition, the histopathological study confirmed the perturbation of biochemical parameters and showed that the hepatic and renal structures were altered. Indeed, the hepatotoxic and nephrotoxic effects are more pronounced when cheese was exposed to the sun.

Threat of plastic ageing in marine environment. Adsorption/desorption of micropollutants.

Ageing of various plastics in marine environment was monitored after immersion of two synthetic (polyvinylchloride, PVC, and polyethylene terephthalate, PET) and one biodegradable (poly(butylene adipate co-terephtalate), PBAT) plastics for 502days in the bay of Lorient (Brittany, France). Data analysis indicates that aged PVC rapidly releases estrogenic compounds in seawater with a later adsorption of heavy metals; PET undergoes a low weakening of the surface whereas no estrogenic activity is detected; PBAT ages faster in marine environment than PVC. Aged PBAT exhibits heterogeneous surface with some cavities likely containing clay minerals from the chlorite group. Besides, this degraded material occasionally shows a high estrogenic activity. Overall, this study reports, for the first time, that some aged plastics, without being cytotoxic, can release estrogenic compounds in marine environment.