Comparison the functional properties of protein hydrolysates from poultry by-products and rainbow trout (Onchorhynchus mykiss) viscera

Poultry by-products and rainbow trout (Onchorhynchus mykiss) viscera are abundant and underutilized resources that can be used as a unique protein source to make protein hydrolysates. In this study protein hydrolysate were made from these two different sources with Alcalase 2.4L. The functional properties of Fish viscera protein hydrolysate (FPH) compared to poultry by-products protein hydrolysate (PPH) were studied: Solubility, water holding capacity (WHC), oil absorption capacity (OAC), colour, emulsifying and foaming properties. Furthermore, the products were characterized by analyzing their amino acid composition. WHC, emulsifying activity, emulsifying stability and foaming properties and color of the FPH was significantly (P 0.05) different. Methionine and histidine in both protein hydrolysates were the limiting amino acids and FPH had more hydrophobic residue. The differences in the amino acid composition between PPH and FPH may also be responsible for their different behaviours at various pH

On-line tracking of the human gut microbial metabolism: high-throughput screening during colonic in-vitro fermentation

The human gut encloses a large community of bacteria producing a wide range of volatile organic compounds (VOCs) when fermenting undigestible substrates. This study aims to provide a high throughput method to study in real-time the gut microbial volatilome when the microbiota process undigestible dietary substrates. Background: Small metabolites from the human gut microbiota are recognized as the intermediates of the microbiome-host cross-talk [1]. The research on the human gut metabolome is mainly based on discrete sampling representing discontinuous ‘snapshot’ of these complex biological systems [2]. The aim of this research work is to enhance the current understanding of the dynamics of the gut microbiota by integrating non-invasive and continuous analytical methods with in-vitro gut simulators, to monitor in real-time, the progression of small molecules released into the headspace [2,3] Methodology: Automated Head space-Solid Phase Micro Extraction coupled with Gas Chromatography-Mass Spectrometry (HS-SPME-GC-MS) and Static Headspace- Proton Transfer Reaction-Time of Flight-Mass Spectrometry (SHS-PTR-ToF-MS) are used for the purpose of this investigation. The objective is to screen and monitor a specific set of masses of interest, to gain system level mechanistic insights on primary metabolism of the gut microbial consortia. Results: This methodology enabled the continuous monitoring of multiple metabolites in time, including short-chain fatty acids (SCFAs) and medium-chain fatty acids (MCFAs) derived from 24h oat bran fermentation. A mixture of -odd and -even chain acids were co-released into the culture headspace after 4 hours of fermentation and their relative abundance increased in time over 24 hours. The production of multiple MCFAs from the substrate is most likely a community optimization strategy to maximize ATP production from oat degradation by means of reverse beta-oxidation which involves the utilization of fermentation intermediates, such as propanol and acetate. Furthermore, the untargeted screening allowed the detection of low abundant sulfur metabolites, thiophenes, which, to our knowledge, were never investigated before as gut microbial metabolites (GMMs). Conclusion: By integrating non-invasive and continuous analytical methods with an in-vitro gut simulator, it was possible to monitor in real-time the progression of two important class of small molecules released by the microbial consortia into the headspace. The collected information can be jointly integrated to shed light on the dynamics of bacterial foraging of complex undigestible substrates (e.g. bran from cereals). Overall, these results confirm the idea to consider the bacterial headspace as a highly dynamic chemical system that contains information on microbial community behavio

Specific Polyunsaturated Fatty Acids Can Modulate in vitro Human moDC2s and Subsequent Th2 Cytokine Release

Allergy is becoming a rapidly increasing problem worldwide, and in vitro models are frequently used to study the mechanisms behind the different types of allergic response. The dendritic cell (DC)\u2013T-cell model can be used to study sensitization. However, lipopolysaccharide (LPS) is often used to maturate the DCs, but it gives rise to a DC1 phenotype, whereas Th2-driven inflammatory diseases such as allergy are characterized by the involvement of the DC2 phenotype. Our aim was to create a DC2\u2013T-cell human model (human moDC2s) to study in vitro sensitization and validate the model using polyunsaturated fatty acids (PUFAs) that were previously shown to have immunomodulatory properties. We found that the generated DC2s expressed OX40L and drove naive T-cells into IL-13 production of CD4+ effector T-cells. In line with in vivo findings, n 123 long-chain (LC)PUFA docosahexaenoic acid (DHA) effectively decreased the DC2's surface expression of OX40L, as well as the IL-12p40 and IL-23 cytokine production by DC2s and subsequently lowered IL-13 production by DC2-induced effector T-cells. Similar cytokine production effects were found with eicosapentaenoic acid (EPA) and arachidonic acid (AA), whereas linoleic acid (LA) increased OX40L surface expression and subsequent T-cell-derived IL-13/IFN\u3b3 ratios, suggesting an increased risk of allergy development. Altogether, these data show that human moDC2s are able to induce Th2-type IL-13 secretion by T-cell differentiated in the presence of these DC2s and that this model can be differentially modulated by PUFAs. These results are in line with previous in vivo studies using PUFAs, indicating that this model may be of use to predict in vivo outcomes

A viral chitinase enhances oral activity of TMOF

In this study we investigate the combined effect on Heliothis virescens (Lepidoptera, Noctuidae) larvae of Aedes aegypti-Trypsin Modulating Oostatic Factor (. Aea-TMOF), a peptide that inhibits trypsin synthesis by the gut, impairing insect digestive function, and Autographa californica nucleopolyhedrovirus Chitinase A (AcMNPV ChiA), an enzyme that is able to alter the permeability of the peritrophic membrane (PM). Aea-TMOF and AcMNPV ChiA were provided to the larvae by administering transgenic tobacco plants, co-expressing both molecules. Experimental larvae feeding on these plants, compared to those alimented on plants expressing only one of the two molecules considered, showed significantly stronger negative effects on growth rate, developmental time and mortality. The impact of AcMNPV ChiA on the PM of H. virescens larvae, measured as increased permeability to molecules, was evident after five days of feeding on transgenic plants expressing ChiA. This result was confirmed by in vitro treatment of PM with recombinant ChiA, extracted from the transgenic plants used for the feeding experiments. Collectively, these data indicate the occurrence of a positive interaction between the two transgenes concurrently expressed in the same plant. The hydrolytic activity of ChiA on the PM of tobacco budworm larvae enhances the permeation of TMOF molecules to the ectoperitrophic space, and its subsequent absorption. The permeation through the paracellular route of Aea-TMOF resulted in a spotted accumulation on the basolateral domain of enterocytes, which suggests the occurrence of a receptor on the gut side facing the haemocoel. The binding of the peptide, permeating at increased rates due to the ChiA activity, is considered responsible for the enhanced insecticide activity of the transgenic plants expressing both molecules. These data corroborate the idea that ChiA can be effectively used as gut permeation enhancer in oral delivery strategies of bioinsecticides targeting haemocoelic receptors

Sugar and cocoa: sweet synergy or bitter antagonisms. Formulating cocoa and chocolate products for health: a narrative review

The potential health effects of cocoa flavanols are well described. Ranging from reducing risk of developing type 2 diabetes and cardiovascular disease at population levels, moderating disease risk factors including endothelial function and lipid metabolism in clinical trials and mechanistic studies in laboratory studies highlighting target tissues and pathways. However, translating these benefits into public health messages is problematic, due to the high energy and sugar content of many cocoa products, including chocolate. This review considered the role of sugar in cocoa products, what are its physiological effects on bioavailability and bioactivity? Considering, then how cocoa products can be reformulated to reduce sugar intake, and the likely effects on beneficial effects of cocoa flavanols and consumer preferences. Ultimately, although interesting physiological effects are seen with cocoa flavanols, their use as a disease‐modifying commodities may be limited the effect such products may have within an individual's and populations overall dietary patterns

tabAnti-HER2 (erbB-2) oncogene effects of phenolic compounds directly isolated from commercial Extra-Virgin Olive Oil (EVOO)

<p>Abstract</p> <p>Background</p> <p>The effects of the olive oil-rich Mediterranean diet on breast cancer risk might be underestimated when HER2 (<it>ERB</it>B2) oncogene-positive and HER2-negative breast carcinomas are considered together. We here investigated the anti-HER2 effects of phenolic fractions directly extracted from Extra Virgin Olive Oil (EVOO) in cultured human breast cancer cell lines.</p> <p>Methods</p> <p>Solid phase extraction followed by semi-preparative high-performance liquid chromatography (HPLC) was used to isolate phenolic fractions from commercial EVOO. Analytical capillary electrophoresis coupled to mass spectrometry was performed to check for the composition and to confirm the identity of the isolated fractions. EVOO polyphenolic fractions were tested on their tumoricidal ability against HER2-negative and HER2-positive breast cancer <it>in vitro </it>models using MTT, crystal violet staining, and Cell Death ELISA assays. The effects of EVOO polyphenolic fractions on the expression and activation status of HER2 oncoprotein were evaluated using HER2-specific ELISAs and immunoblotting procedures, respectively.</p> <p>Results</p> <p>Among the fractions mainly containing the <it>single phenols </it>hydroxytyrosol and tyrosol, the <it>polyphenol acid </it>elenolic acid, the <it>lignans </it>(+)-pinoresinol and 1-(+)-acetoxypinoresinol, and the <it>secoiridoids </it>deacetoxy oleuropein aglycone, ligstroside aglycone, and oleuropein aglycone, all the major EVOO polyphenols (<it>i.e. </it>secoiridoids and lignans) were found to induce strong tumoricidal effects within a micromolar range by selectively triggering high levels of apoptotic cell death in HER2-overexpressors. Small interfering RNA-induced depletion of HER2 protein and lapatinib-induced blockade of HER2 tyrosine kinase activity both significantly prevented EVOO polyphenols-induced cytotoxicity. EVOO polyphenols drastically depleted HER2 protein and reduced HER2 tyrosine autophosphorylation in a dose- and time-dependent manner. EVOO polyphenols-induced HER2 downregulation occurred regardless the molecular mechanism contributing to HER2 overexpression (<it>i.e</it>. naturally by gene amplification and ectopically driven by a viral promoter). Pre-treatment with the proteasome inhibitor MG132 prevented EVOO polyphenols-induced HER2 depletion.</p> <p>Conclusion</p> <p>The ability of EVOO-derived polyphenols to inhibit HER2 activity by promoting the proteasomal degradation of the HER2 protein itself, together with the fact that humans have safely been ingesting secoiridoids and lignans as long as they have been consuming olives and OO, support the notion that the stereochemistry of these phytochemicals might provide an excellent and safe platform for the design of new HER2-targeting agents.</p

Olive oil's bitter principle reverses acquired autoresistance to trastuzumab (Herceptin™) in HER2-overexpressing breast cancer cells

[Background] A low incidence of breast cancer in the Mediterranean basin suggests that a high consumption of Extra Virgin Olive Oil (EVOO) might confer this benefit. While the anti-HER2 oncogene effects of the main ω-9 fatty acid present in EVOO triacylglycerols (i.e., oleic acid) have been recently described, the anti-breast cancer activities of EVOO non-glyceridic constituents -which consist of at least 30 phenolic compounds-, remained to be evaluated. [Methods] Semi-preparative HPLC was used to isolate EVOO polyphenols (i.e., tyrosol, hydroxytyrosol, oleuropein). Both the anti-proliferative and the pro-apoptotic effects of EVOO phenolics were evaluated by using MTT-based quantification of metabolically viable cells and ELISA-based detection of histone-associated DNA fragments, respectively. The nature of the interaction between oleuropein aglycone and the anti-HER2 monoclonal antibody trastuzumab (Herceptin™) was mathematically evaluated by the dose-oriented isobologram technique. HER2-specific ELISAs were employed to quantitatively assess both the basal cleavage of the HER2 extracellular domain (ECD) and the expression level of total HER2. The activation status of HER2 was evaluated by immunoblotting procedures using a monoclonal antibody specifically recognizing the tyrosine phosphorylated (Phosphor-Tyr1248) form of HER2. [Results] Among EVOO polyphenols tested, oleuropein aglycone was the most potent EVOO phenolic in decreasing breast cancer cell viability. HER2 gene-amplified SKBR3 cells were ~5-times more sensitive to oleuropein aglycone than HER2-negative MCF-7 cells. Retroviral infection of the HER2 oncogene in MCF-7 cells resulted in a "SKBR3-assimilated" phenotype of hypersensitivity to oleuropein aglycone. An up to 50-fold increase in the efficacy of trastuzumab occurred in the presence of oleuropein aglycone. A preclinical model of acquired autoresistance to trastuzumab (SKBR3/Tzb100 cells) completely recovered trastuzumab sensitivity (> 1,000-fold sensitization) when co-cultured in the presence of oleuropein aglycone. Indeed, the nature of the interaction between oleuropein aglycone and trastuzumab was found to be strongly synergistic in Tzb-resistant SKBR3/Tzb100 cells. Mechanistically, oleuropein aglycone treatment significantly reduced HER2 ECD cleavage and subsequent HER2 auto-phosphorylation, while it dramatically enhanced Tzb-induced down-regulation of HER2 expression. [Conclusion] Olive oil's bitter principle (i.e., oleuropein aglycone) is among the first examples of how selected nutrients from an EVOO-rich "Mediterranean diet" directly regulate HER2-driven breast cancer disease.JAM is the recipient of a Basic, Clinical and Translational Research Award (BCTR0600894) from the Susan G. Komen Breast Cancer Foundation (Texas, USA). This work was also supported by the Instituto de Salud Carlos III (Ministerio de Sanidad y Consumo, Fondo de Investigación Sanitaria -FIS-, Spain, Grants CP05-00090 and PI06-0778 to JAM, and Grant RD06-0020-0028 to JAM, RC and JB)