Predicting the Oxidative Degradation of Raw Beef Meat during Cold Storage Using Numerical Simulations and Sensors-Prospects for Meat and Fish Foods.

Preventing animal-source food waste is an important pathway to reducing malnutrition and improving food system sustainability. Uncontrolled color variation due to oxidation is a source of waste as it prompts food rejection by consumers. Evaluation of oxidation-reduction potential (ORP) can help to predict and prevent oxidation and undesirable color changes. A new sensor and two modeling approaches-a phenomenological model and a reaction-diffusion model-were successfully used to predict the oxidative browning of beef ribeye steaks stored under different temperature and oxygen concentration conditions. Both models predicted similar storage durations for acceptable color, although deviating for higher and lower redness levels, which are of no interest for meat acceptance. Simulations under higher oxygen concentrations lead to a few days of delay in the redness change, as observed in practice, under modified atmosphere packaging. In meat juice, variation in ORP measured by the sensor correlated with the redness variation. However, in meat, sensors promote oxidation in the adjacent area, which is unacceptable for industrial use. This paper discusses the potential, limits, and prospects of the mathematical models and sensors, developed for beef. A strategy is proposed to couple these approaches and include the effect of microorganisms.

Digging Deeper into Breast Cancer Epigenetics: Insights from Chemical Inhibition of Histone Acetyltransferase TIP60 In Vitro.

Breast cancer is often sporadic due to several factors. Among them, the deregulation of epigenetic proteins may be involved. TIP60 or KAT5 is an acetyltransferase that regulates gene transcription through the chromatin structure. This pleiotropic protein acts in several cellular pathways by acetylating proteins. RNA and protein expressions of TIP60 were shown to decrease in some breast cancer subtypes, particularly in triple-negative breast cancer (TNBC), where a low expression of TIP60 was exhibited compared with luminal subtypes. In this study, the inhibition of the residual activity of TIP60 in breast cancer cell lines was investigated by using two chemical inhibitors, TH1834 and NU9056, first on the acetylation of the specific target, lysine 4 of histone 3 (H3K4) by immunoblotting, and second, by chromatin immunoprecipitation (ChIP)-qPCR (-quantitative Polymerase Chain Reaction). Subsequently, significant decreases or a trend toward decrease of H3K4ac in the different chromatin compartments were observed. In addition, the expression of 48 human nuclear receptors was studied with TaqMan Low-Density Array in these breast cancer cell lines treated with TIP60 inhibitors. The statistical analysis allowed us to comprehensively characterize the androgen receptor and NR3C2 receptors in TNBC cell lines after TH1834 or NU9056 treatment. The understanding of the residual activity of TIP60 in the evolution of breast cancer might be a major asset in the fight against this disease, and could allow TIP60 to be used as a biomarker or therapeutic target for breast cancer progression in the future.

Polyethyleneimine as a tool for compounds fractionation by flocculation in a microalgae biorefinery context.

In the context of emerging biorefinery for microalgae, polyethyleneimine (PEI), has been tested in order to achieve separation of fat-soluble and water-soluble compounds from Haematococcus pluvialis. Several parameters were taken into account (ratio between sample and PEI, pH, and ionic strength) and 2 conditions (0.075% PEI pH 7.4, and 0.100% PEI pH8.5) were studied for up-scalability, with a recovery of flocculated compounds (lipids and pigments), and a complete characterization of both phases. Using 0.075% PEI, pH7.4, 100% sugars and 89.8% proteins were retained in the supernatant, but some trace of beta-carotene were also detected. For 0.100% PEI, pH 8.5, a loss in proteins content was highlighted (61.2% proteins retained), but no residual lipids or pigments were detected. PEI could therefore be considered as an efficient method to fractionate fat-soluble and water-soluble compounds from microalgae.

Prediction of pH and aw of pork meat by a thermodynamic model: New developments.

To ensure continuous innovations, food industries need tools which enable to predict physical-properties of food during a change of process or recipe. In this work, a thermodynamic model is suggested to predict pH and water activity of pork meat in presence of different additives such as salts or organic acids used in food industry. The predictions of pH and aw are satisfactory in a wide prediction domain, with a good accuracy. In add, a neural network mimetic of thermodynamic model is developed in order to facilitate the use of thermodynamic model and reduce calculation time.

Impact of reducing nitrate/nitrite levels on the behavior of Salmonella Typhimurium and Listeria monocytogenes in French dry fermented sausages.

Salmonella and Listeria monocytogenes are two pathogenic bacteria that most frequently contaminate pork meat. In dry fermented sausages, several hurdles are used for controlling bacterial growth such as nitrite and salt addition. In Europe, practices consist of adding potassium nitrate (250ppm expressed as NaNO3) or a combination of nitrate/nitrite (150/150ppm expressed as NaNO3/NaNO2 respectively). However, involvement of these additives in nitrosamine formation is a matter of concern. Consequently, a decrease in nitrite/nitrate amounts is proposed. The aim of this study was to evaluate the impact of reducing levels of these additives on Listeria and Salmonella behavior. Using challenge-tests, five trials were carried out by varying the concentration of nitrate and nitrate/nitrite. Results shown that nitrite is a relevant hurdle for control Salmonella and Listeria. At the end of drying, the most significant reductions of pathogens are obtained in sausages with nitrite added at the both tested concentrations (120 or 80ppm NaNO2).

The Epigenetic Landscape of Promoter Genome-wide Analysis in Breast Cancer.

Breast cancer is a heterogeneous disease due to its clinico-pathological features and response to therapy. The classification of breast tumors based on their hormone receptor status and pathologic features. Post-translational histone modifications come into prominence for regulation of gene expression in cancer pathogenesis. Here, we analyzed dysregulation of H3K9ac and H3K27me3-enriched subtype-specific genes using ChIP-on-chip assay in breast cancer tumors and matched normal tissue samples. Breast cancer tumors were classified according to St Gallen Consensus 2013. Our results indicated that the promoter regions of genes modified by H3K9ac epi-mark are commonly associated with tumors with HER2-positive and TNBC subtype. H3K27me3-enriched genes were comprised of Luminal A and B1 subtypes. We constructed a network structure to elicit epigenetically regulated genes related with breast cancer progression. The central genes of the network (RUNX1, PAX3, GATA4 and DLX5) were subjected for epigenetically dysregulation in association with different breast cancer subtypes. Our study submits epigenetic mechanisms are crucial to elicit subtype-specific regulation in breast cancer and ChIP-on-chip assay provides a better understanding for breast tumorigenesis and new approaches for prevention and treatment.

Global analysis of H3K27me3 as an epigenetic marker in prostate cancer progression.

BACKGROUND: H3K27me3 histone marks shape the inhibition of gene transcription. In prostate cancer, the deregulation of H3K27me3 marks might play a role in prostate tumor progression. METHODS: We investigated genome-wide H3K27me3 histone methylation profile using chromatin immunoprecipitation (ChIP) and 2X400K promoter microarrays to identify differentially-enriched regions in biopsy samples from prostate cancer patients. H3K27me3 marks were assessed in 34 prostate tumors: 11 with Gleason score > 7 (GS > 7), 10 with Gleason score ≤ 7 (GS ≤ 7), and 13 morphologically normal prostate samples. RESULTS: Here, H3K27me3 profiling identified an average of 386 enriched-genes on promoter regions in healthy control group versus 545 genes in GS ≤ 7 and 748 genes in GS > 7 group. We then ran a factorial discriminant analysis (FDA) and compared the enriched genes in prostate-tumor biopsies and normal biopsies using ANOVA to identify significantly differentially-enriched genes. The analysis identified ALG5, EXOSC8, CBX1, GRID2, GRIN3B, ING3, MYO1D, NPHP3-AS1, MSH6, FBXO11, SND1, SPATS2, TENM4 and TRA2A genes. These genes are possibly associated with prostate cancer. Notably, the H3K27me3 histone mark emerged as a novel regulatory mechanism in poor-prognosis prostate cancer. CONCLUSIONS: Our findings point to epigenetic mark H3K27me3 as an important event in prostate carcinogenesis and progression. The results reported here provide new molecular insights into the pathogenesis of prostate cancer.

H3K4 acetylation, H3K9 acetylation and H3K27 methylation in breast tumor molecular subtypes.

AIM: Here, we investigated how the St Gallen breast molecular subtypes displayed distinct histone H3 profiles. PATIENTS & METHODS: 192 breast tumors divided into five St Gallen molecular subtypes (luminal A, luminal B HER2-, luminal B HER2+, HER2+ and basal-like) were evaluated for their histone H3 modifications on gene promoters. RESULTS: ANOVA analysis allowed to identify specific H3 signatures according to three groups of genes: hormonal receptor genes (ERS1, ERS2, PGR), genes modifying histones (EZH2, P300, SRC3) and tumor suppressor gene (BRCA1). A similar profile inside high-risk cancers (luminal B [HER2+], HER2+ and basal-like) compared with low-risk cancers including luminal A and luminal B (HER2-) were demonstrated. CONCLUSION: The H3 modifications might contribute to clarify the differences between breast cancer subtypes.

Epigenetic Modifications with DZNep, NaBu and SAHA in Luminal and Mesenchymal-like Breast Cancer Subtype Cells.

BACKGROUND/AIM: Numerous studies have shown that breast cancer and epigenetic mechanisms have a very powerful interactive relation. The MCF7 cell line, representative of luminal subtype and the MDA-MB 231 cell line representative of mesenchymal-like subtype were treated respectively with a Histone Methyl Transferase Inhibitors (HMTi), 3-Deazaneplanocin hydrochloride (DZNep), two histone deacetylase inhibitors (HDACi), sodium butyrate (NaBu), and suberoylanilide hydroxamic acid (SAHA) for 48 h. MATERIALS AND METHODS: Chromatin immunoprecipitation (ChIP) was used to observe HDACis (SAHA and NaBu) and HMTi (DZNep) impact on histones and more specifically on H3K27me3, H3K9ac and H3K4ac marks with Q-PCR analysis of BRCA1, SRC3 and P300 genes. Furthermore, the HDACi and HMTi effects on mRNA and protein expression of BRCA1, SRC3 and P300 genes were checked. In addition, statistical analyses were used. RESULTS: In the MCF7 luminal subtype with positive ER, H3k4ac was significantly increased on BRCA1 with SAHA. On the contrary, in the MDA-MB 231 breast cancer cell line, representative of mesenchymal-like subtype with negative estrogen receptor, HDACis had no effect. Also, DZNEP decreased significantly H3K27me3 on BRCA1 in MDA-MB 231. Besides, on SRC3, a significant increase for H3K4ac was obtained in MCF7 treated with SAHA. And DZNEP had no effect in MCF7. Also, in MDA-MB 231 treated with DZNEP, H3K27me3 significantly decreased on SRC3 while H3K4ac was significantly increased in MDA-MB-231 treated with SAHA or NaBu for P300. CONCLUSION: Luminal and mesenchymal-like breast cancer subtype cell lines seemed to act differently to HDACis (SAHA and NaBu) or HMTi (DZNEP) treatments.

Improvement and modeling of culture parameters to enhance biomass and lipid production by the oleaginous yeast Cryptococcus curvatus grown on acetate.

The improvement of culture parameters for lipid production from acetate as carbon source was investigated using the oleaginous yeast Cryptococcus curvatus. A new pH regulation system dispensing acetate was developed for fed-batch culture and allowed obtaining nearly 80 g/L biomass within 60 h with a maximal growth rate of 0.28 h(-1). A biological model was developed from experimental data. The influence of three C/N ratios of 300, 500 and 900 were tested during a multi-phases process on lipid accumulation. The C/N ratio of 300 was reported to be the most suitable for lipid storage. No significant increase of lipids content was obtained with higher value. A maximal content of 60% DCW of lipid was obtained. The determination of fatty acids profiles of the microbial oils has confirmed that the valorization of acetate by microbial oils production was a promising perspective.

Genome-wide DNA methylation modified by soy phytoestrogens: role for epigenetic therapeutics in prostate cancer?

In prostate cancer, DNA methylation is significantly associated with tumor initiation, progression, and metastasis. Previous studies have suggested that soy phytoestrogens might regulate DNA methylation at individual candidate gene loci and that they play a crucial role as potential therapeutic agents for prostate cancer. The purpose of our study was to examine the modulation effects of phytoestrogens on a genome-wide scale in regards to DNA methylation in prostate cancer. Prostate cancer cell lines DU-145 and LNCaP were treated with 40 µM of genistein and 110 µM of daidzein. DNMT inhibitor 5-azacytidine (2 µM) and the methylating agent budesonide (2 µM) were used to compare their demethylation/methylation effects with phytoestrogens. The regulatory effects of phytoestrogens on DNA methylation were analyzed by using a methyl-DNA immunoprecipitation method coupled with Human DNA Methylation Microarrays (MeDIP-chip). We observed that the methylation profiles of 58 genes were altered by genistein and daidzein treatments in DU-145 and LNCaP prostate cancer cells. In addition, the methylation frequencies of the MAD1L1, TRAF7, KDM4B, and hTERT genes were remarkably modified by genistein treatment. Our results suggest that the modulation effects of phytoestrogens on DNA methylation essentially lead to inhibition of cell growth and induction of apoptosis. Genome-wide methylation profiling reported here suggests that epigenetic regulation mechanisms and, by extension, epigenetics-driven novel therapeutic candidates warrant further consideration in future "omics" studies of prostate cancer.

Quantitative study of the relationships among proteolysis, lipid oxidation, structure and texture throughout the dry-cured ham process.

Temperature, salt and water contents are key processing factors in dry-cured ham production. They affect how proteolysis, lipid oxidation, structure and texture evolve, and thus determine the sensory properties and final quality of dry-cured ham. The aim of this study was to quantify the interrelationships and the time course of (i) proteolysis, (ii) lipid oxidation, (iii) five textural parameters: hardness, fragility, cohesiveness, springiness and adhesiveness and (iv) four structural parameters: fibre numbers, extracellular spaces, cross section area, and connective tissue area, during the dry-cured ham process. Applying multiple polynomial regression enabled us to build phenomenological models relating proteolysis, salt and water contents to certain textural and structural parameters investigated. A linear relationship between lipid oxidation and proteolysis was also established. All of these models and relationships, once combined with salt penetration, water migration and heat transfer models, can be used to dynamically simulate all of these phenomena throughout dry-cured ham manufacturing.

The association between histone 3 lysine 27 trimethylation (H3K27me3) and prostate cancer: relationship with clinicopathological parameters.

BACKGROUND: It is well established that genetic and epigenetic alterations are common events in prostate cancer, which may lead to aberrant expression of critical genes. The importance of epigenetic mechanisms in prostate cancer carcinogenesis is increasingly evident. In this study, the focus will be on histone modifications and the primary objectives are to map H3K27me3 marks and quantify RAR beta 2, ER alpha, SRC3, RGMA, PGR, and EZH2 gene expressions in prostate cancer tissues compared to normal tissues. In addition, a data analysis was made in connection with the clinicopathological parameters. METHODS: 71 normal specimens and 66 cancer prostate tissues were randomly selected in order to assess the proportion of the repressive H3K27me3 mark and gene expression. H3K27me3 level was evaluated by ChIP-qPCR and mRNA expression using RT-qPCR between prostate cancer and normal tissues. Subsequently, western-blotting was performed for protein detection. The analysis of variance (ANOVA) was performed, and Tukey's test was used to correct for multiple comparisons (p-value threshold of 0.05). The principal component analysis (PCA) and discriminant factorial analysis (DFA) were used to explore the association between H3K27me3 level and clinicopathological parameters. RESULTS: The study demonstrated that H3K27me3 level was significantly enriched at the RAR beta 2, ER alpha, PGR, and RGMA promoter regions in prostate cancer tissues compared to normal tissues. After stratification by clinicopathological parameters, the H3K27me3 level was positively correlated with Gleason score, PSA levels and clinical stages for RAR beta 2, ER alpha, PGR, and RGMA. High H3K27me3 mark was significantly associated with decreased RAR beta 2, ER alpha, PGR and RGMA gene expressions in prostate cancer sample compared to the normal one. Moreover, the results showed that mRNA level of EZH2, AR and SRC3 are upregulated in prostate cancer compared to normal prostate tissues and this correlates positively with Gleason score, PSA levels and clinical stages. Obviously, these observations were confirmed by protein level using western-blot. CONCLUSIONS: This data clearly demonstrated that H3K27me3 level correlated with aggressive tumor features. Also this study revealed that reverse correlation of RAR beta 2, ER alpha, PGR, and RGMA expressions with EZH2, SRC3, and AR expressions in prostate cancer tissues suggests that these genes are the target of EZH2. Therefore, all therapeutic strategies leading to histone demethylation with epigenetic drugs such as histone methyltransferase inhibitor may be relevant treatments against prostate cancer.

Comparative effects of soy phytoestrogens and 17ß-estradiol on DNA methylation of a panel of 24 genes in prostate cancer cell lines.

Major phytoestrogens genistein and daidzein have been reported to have the ability to reverse DNA methylation in cancer cell lines. The mechanism by which genistein and daidzein have an inhibiting action on DNA methylation is not well understood. The aim of this study was to investigate the effects of soy phytoestrogens and the natural estrogen 17ß-estradiol (E2) to determine whether one of the estrogen receptors is mobilized for the action of these compounds on DNA methylation. We also made a comparative study with a DNA methylation inhibitor (5-azacytidine) and a DNA methylation activator (budesonide). Three prostate cell lines, PC-3, DU-145, and LNCaP, were treated with 40 µM genistein, 110 µM daidzein, 2 µM budesonide, 2 µM 5-azacytidine, and 10 µM E2. In these 3 human prostate cancer cell lines, we performed methylation quantification using methyl-profiler-DNA-methylation analysis. Soy phytoestrogens and E2 induced a demethylation of all the promoter regions studied except for those that were unmethylated in control cells. Our results showed that E2 induces, like soy phytoestrogen, a decrease in DNA methylation in prostate cancer cell lines. This action may be mediated through ERß.

Building phenomenological models that relate proteolysis in pork muscles to temperature, water and salt content.

Throughout dry-cured ham production, salt and water content, pH and temperature are key factors affecting proteolysis, one of the main biochemical processes influencing sensory properties and final quality of the product. The aim of this study was to quantify the effect of these variables (except pH) on the time course of proteolysis in laboratory-prepared pork meat samples. Based on a Doehlert design, samples of five different types of pork muscle were prepared, salted, dried and placed at different temperatures, and sampled at different times for quantification of proteolysis. Statistical analysis of the experimental results showed that the proteolysis index (PI) was correlated positively with temperature and water content, but negatively with salt content. Applying response surface methodology and multiple linear regressions enabled us to build phenomenological models relating PI to water and salt content, and to temperature. These models could then be integrated into a 3D numerical ham model, coupling salt and water transfers to proteolysis.

Modelling the distribution of a(w), pH and ions in marinated beef meat.

New beef products from low value cuts could be developed using marinating since this process has been shown to improve meat sensorial properties and shelf life. However, to optimise the process mathematical models are needed to predict evolution of the physicochemical properties that determine biochemical and structural changes. Two major works have been carried out to elaborate comprehensive models: (1) Thermodynamic models were adapted to predict water sorption isotherms and pH of beef meat tissue in presence of salts (NaCl, KCl) and organic acids (acetic, lactic, citric and ascorbic acid), (2) Fickian numerical models were set up to predict the migration of ions within meat cuts using apparent diffusivities previously estimated from 1D experiments. Simulation calculations showed reasonable agreement with measurements and can be used to investigate the effect of marinating conditions, product heterogeneity, dimension and shape.

Development of an enzymatic method to quantify methyl ketones from bacterial origin.

Methyl ketones are detected in dry fermented sausages in which they contribute to the cured aroma. They have been associated with the inoculation of Staphylococcus carnosus used as starter culture. To evaluate the ability of bacterial starters to produce methyl ketones it was necessary to develop a rapid method. The method consists of a reaction catalyzed by a commercial NADPH-dependent alcohol dehydrogenase that reduces the 2-pentanone to its secondary alcohol. The linearity, the specificity, and the robustness were studied. Its accuracy was confirmed by comparison with the gas chromatography technique. Finally, the method was validated on biological samples such as the 2-pentanone produced by Staphylococcus carnosus. The enzymatic method offers some advantages over the gas chromatography, as it is faster, simpler, and inexpensive, guaranteeing an effective way to assess bacterial ketone production.