Grape pomace enzymatic extract restores vascular dysfunction evoked by endothelin-1 and DETCA via NADPH oxidase downregulation and SOD activation

Grape pomace enzymatic extract (GP-EE) exhibits a potent antioxidant and vasodilator activity, although the mechanisms underlying this action are not completely understood. Our aim was to evaluate its vasoactive mechanisms, focusing on vascular NADPH oxidase and superoxide dismutase (SOD)-dependent pathways. GP-EE effectively restored vascular impairment induced by the NADPH oxidase activator endothelin-1 through downregulation of its Nox-1 and p47^phox subunits. GP-EE partially prevented endothelial dysfunction elicited by the SOD inhibitor, DETCA, preserving EC-SOD and Mn-SOD isoenzymes protein expression. Although many of these vasoactive effects could be attributed to the presence of catechin, GP-EE was able to exert additional protection against the vascular deleterious effects evoked by NADPHox activation and SOD inhibition. This additional vasoprotection could be related to the enzymatic extraction process, providing polyphenols from winemaking residue with a potent activity against vascular dysfunction by preventing NADPHox activation and maintaining SOD defense, making GP-EE as an ideal nutritional supplement.     

Factors effecting dental composite wear in vitro

The dependence on pH of the absorption and circular dichroic spectra of iron(III), cobalt(III) and copper(II) transferrins has been (re)investigated. In the alkaline region, the CD profiles of iron(III) and cobalt(III) transferrin are essentially pH independent up to pH 11; only for very high pH values (pH > 11) is breakdown of the cobalt(III) and iron(III) transferrin derivatives observed, without evidence of conformational rearrangements. By contrast, the CD profiles of copper transferrin show drastic changes in shape around pH 10; these spectral changes, which are fitted to a pKa of approximately 10.4, are interpreted in terms of a substantial rearrangement of the local environment of the copper ions at high pH. Although the CD spectra of copper transferrin at alkaline pH strictly resemble those observed upon addition of modifier anions, the mechanism of site destabilization in the two cases is different; at variance with the case of modifier anions, our results suggest that the high pH form of copper transferrin still contains the synergistic anion. A 13C NMR experiment has confirmed this view. In the acidic region, iron(III) and cobalt(III) transferrins are stable down to pH approximately 6. For lower pH values progressive metal detachment is observed without evidence of conformational changes; around pH 4.5 most bound metals are released. In the case of the less stable copper-transferrin, metal removal from the specific binding sites is already complete around pH 6.0; in concomitance with release from the primary sites, binding of copper ions to secondary sites is observed. Additional information has been gained from CD experiments in the far UV. The pH dependent properties of iron(III), cobalt(III) and copper(II) transferrin are discussed in the frame of the present knowledge of transferrin chemistry, particular emphasis being attributed to the comparison between tripositive and bipositive metal derivatives.     

Physicochemical,nutritional and functional characteristics of two underutilised fruit cactus species (Myrtillocactus) produced in central Mexico

The physicochemical, nutritional, and functional variations of Myrtillocactus geometrizans and Myrtillocactus schenckii, were assessed. Variations of characteristics among species and geographic origin were lower compared to prickly pear. Titratable acidity, and total soluble solids varied by 7.7%, and 11%, respectively. Variations of protein (18.5%) and fibre (5.2%) contents were lower when compared to prickly pear. Iron and copper contents make Myrtillocactus a good source of minerals contributing 16% and 20% of daily requirements. Myrtillocactus also could be an outstanding source of fibre (up to 36.9%). Gallic and caffeic acids, and vanillin were identified by HPLC. Betalains, soluble phenolics, and vitamin C contribute to the high antioxidant activity (TEAC) which, if compared with commonly consumed fruits, was higher. The potential for the production of high-quality industrialised products derived from Myrtillocactus is feasible, as long as compositional variations from batch to batch are kept in check.     

Management of Acute Lung Injury: Palmitoylethanolamide as a New Approach

Acute lung injury (ALI) and acute respiratory distress syndrome (ARDS) are common and devastating clinical disorders with high mortality and no specific therapy. Lipopolysaccharide (LPS) is usually used intratracheally to induce ALI in mice. The aim of this study was to examine the effects of an ultramicronized preparation of palmitoylethanolamide (um-PEA) in mice subjected to LPS-induced ALI. Histopathological analysis reveals that um-PEA reduced alteration in lung after LPS intratracheal administration. Besides, um-PEA decreased wet/dry weight ratio and myeloperoxidase, a marker of neutrophils infiltration, macrophages and total immune cells number and mast cells degranulation in lung. Moreover, um-PEA could also decrease cytokines release of interleukin (IL)-6, interleukin (IL)-1β, tumor necrosis factor (TNF)-α and interleukin (IL)-18. Furthermore, um-PEA significantly inhibited the phosphorylation of nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor, alpha (IκBα) and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) activation in ALI, and at the same time decreased extracellular signal-regulated kinase 1/2 (ERK1/2), c-Jun N-terminal kinase (JNK) and p38 mitogen-activated protein kinase (p38/MAPK) expression, that was increased after LPS administration. Our study suggested that um-PEA contrasted LPS-induced ALI, exerting its potential role as an adjuvant anti-inflammatory therapeutic for treating lung injury, maybe also by p38/NF-κB pathway.     

Multitoxin extraction and detection of trichothecenes in cereals: an improved LC‐MS/MS approach

BACKGROUND: Many multiresidual methods to evaluate natural occurrence of Fusarium toxins are already reported in the scientific literature but a new rapid, reliable, cost‐efficient and high‐sensitivity method for the simultaneous determination of several fusariotoxins is always welcome. Nivalenol (NIV), deoxynivalenol, fusarenon‐X (FUS‐X), 3‐acetyldeoxynivalenol, diacetoxyscirpenol (DAS), HT‐2 toxin, T‐2 toxin, neosolaniol (NEO), zearalanone and zearalenone (ZON) belong to the most common mycotoxins in food matrix grains, e.g., wheat and maize. The proposed method is a multitoxin analytical method that combines high‐performance liquid chromatography (HPLC), atmospheric pressure chemical ionization (APCI), triple‐quadrupole tandem mass spectrometry (LC‐MS/MS) under the selected reaction monitoring (SRM) mode, and it is focused on the optimization of the sample preparation without the need for any cleanup. RESULTS: Three different methods for sample preparation and for the simultaneous extractions of the above‐mentioned fusariotoxins were tested: two of these were followed by a different cleanup step for comparison, while the extraction method proposed in this work, which uses an 84% (v/v) acetonitrile aqueous solution, sample homogenization and subsequent filtration, was validated without any further cleanup step. CONCLUSION: Calibration curves for all analytes are linear, except DAS, HT‐2 and ZON, over the working range of 10–1000 µg kg^?1. The calibration curve of DAS was linear between 10 and 500 µg kg^?1, although the curves of HT‐2 and ZON were linear in the range 10–250 µg kg^?1. Squared correlation coefficients (R²) were in the range 0.995–0.998 for the all point calibration curves. The lowest limits of detection (LOD) were found for DON and ZAN with 0.5 and 0.2 µg kg^?1, respectively, while the highest LODs were obtained for NIV, FUS‐X and NEO, with 3.3 µg kg^?1 for each toxin. Copyright © 2009 Society of Chemical Industry     

Bioprotection of Golden Delicious apples and Iceberg lettuce against foodborne bacterial pathogens by lactic acid bacteria

Lactic acid bacteria were isolated from fresh vegetables and fruit and its ability to inhibit the growth of foodborne human pathogens (Escherichia coli, Listeria monocytogenes, Pseudomonas aeruginosa, Salmonella typhimurium, and Staphylococcus aureus) was tested using the agar spot assay. Eighteen isolates showed a strong antagonistic capacity and were further characterised and identified using 16S rDNA sequencing and API 50CH. Most of them pertained to Leuconostoc spp. and Lactobacillus plantarum, and a few corresponded to Weissella spp. and Lactococcus lactis. Growth and efficacy of control of foodborne pathogen test bacteria by selected strains were tested in wounded Golden Delicious apples and Iceberg lettuce leaf cuts. The strains grew on the substrates and did not cause negative effects on the general aspect of tissues of apple or lettuce. Treatment of apple wounds and lettuce cuts with the antagonistic strains reduced the cell count of S. typhimurium and E .coli by 1 to 2 log cfu/wound or g, whereas the growth of L. monocytogenes was completely inhibited. Results support the potential use of lactic acid bacteria as bioprotective agents against foodborne human pathogens in ready-to-eat fresh fruit and vegetable products.     

Occurrence of mycotoxin in Farro samples from southern Italy

The occurrence of nine mycotoxins and of contamination by pre- and postharvest fungal pathogens of cereals was investigated in samples of stored Triticum monococcum L., Triticum dicoccon Schrank (emmer), and Triticum spelta L. (spelt). In Italy, all three species are collectively referred to as farro. The samples examined were harvested in summer 2000 from eight different sites in southern Italy. Conventional fluorimetric and diode array-based high-performance liquid chromatography (HPLC) analyses and HPLC-mass spectrometry analyses were used to identify fumonisin B1 in five samples (up to 70.00 microg/ kg), ochratoxin A in seven samples (up to 4.07 microg/kg), and beauvericin in three samples (up to 4.44 mg/kg). Enniatin B was detected in one sample (30.00 microg/kg), but no zearalenone or fusaproliferin was found. Deoxynivalenol and aflatoxins were not evaluated. The potentially mycotoxigenic fungal species detected were Alternaria alternata, Fusarium proliferatum, Fusarium tricinctum, Penicillium verrucosum, and Penicillium chrysogenum. This is the first report of the natural occurrence of mycotoxins in farro samples.     

Prevalence of Bacillus spp. in different food products collected in Argentina

The prevalence of Bacillus spp. in 279 samples of different food products collected in Argentina was studied. Bacillus spp. was confirmed in 28 out of 70 honey samples, 29 out of 29 flour samples, 15 out of 50 cheese samples, and 30 out of 30 spice samples, while Bacillus spp. was not found in fresh anchovy. Among the 70 honeys studied, Bacillus cereus, Bacillus pumilus, Bacillus laterosporus and Paenibacillus larvae subspp. larvae showed an incidence of 23%, 4%, 8% and 38%, respectively. More diversity of Bacillus species was found in rye flours than in white flours, Bacillus subtilis being the predominant species isolated from rye flour. B. cereus had an incidence of 50% in Port Salut Argentino cheeses. Meanwhile, B. pumilus was identified in both Port Salut and Quartirolo cheeses with an incidence of 50% and 25%, respectively. All the spices analysed showed Bacillus mycoides as the sole aerobic spore-forming bacilli isolated. The association of the presence of B. cereus, B. subtilis and Bacillus licheniformis with both the potential spoilage of foods and foodborne outbreaks is well known. In this study, Bacillus spp. had an incidence of 38% among all the samples analysed, therefore the monitoring of those species should be routinely done in microbiological food analyses.     

Epigallocatechin-3-Gallate Modulates Postoperative Pain by Regulating Biochemical and Molecular Pathways

Treating postoperative (PO) pain is a clinical challenge. Inadequate PO pain management can lead to worse outcomes, for example chronic post-surgical pain. Therefore, acquiring new information on the PO pain mechanism would increase the therapeutic options available. In this paper, we evaluated the role of a natural substance, epigallocatechin-3-gallate (EGCG), on pain and neuroinflammation induced by a surgical procedure in an animal model of PO pain. We performed an incision of the hind paw and EGCG was administered for five days. Mechanical allodynia, thermal hyperalgesia, and motor dysfunction were assessed 24 h, and three and five days after surgery. At the same time points, animals were sacrificed, and sera and lumbar spinal cord tissues were harvested for molecular analysis. EGCG administration significantly alleviated hyperalgesia and allodynia, and reduced motor disfunction. From the molecular point of view, EGCG reduced the activation of the WNT pathway, reducing WNT3a, cysteine-rich domain frizzled (FZ)1 and FZ8 expressions, and both cytosolic and nuclear β-catenin expression, and the noncanonical β-catenin–independent signaling pathways, reducing the activation of the NMDA receptor subtype NR2B (pNR2B), pPKC and cAMP response element-binding protein (pCREB) expressions at all time points. Additionally, EGCG reduced spinal astrocytes and microglia activation, cytokines overexpression and nuclear factor kappa-light-chain-enhancer of activated B cells (NFkB) pathway, downregulating inducible nitric oxide synthase (iNOS) activation, cyclooxygenase 2 (COX-2) expression, and prostaglandin E2 (PGE2) levels. Thus, EGCG administration managing the WNT/β-catenin signaling pathways modulates PO pain related neurochemical and inflammatory alterations.     

In vitro fermentability and antioxidant capacity of the indigestible fraction of cooked black beans (Phaseolus vulgaris L.), lentils (Lens culinaris L.) and chickpeas (Cicer arietinum L.)

BACKGROUND: Pulses represent an important source of protein, as well as digestible and indigestible carbohydrates. Little information is available on the indigestible carbohydrates and antioxidant capacity of legume seeds. The cooked seeds of three pulses (black bean, chickpea and lentil) were evaluated for their indigestible fraction (IF), polyphenols content, antioxidant capacity and in vitro fermentability, including short‐chain fatty acid production. RESULTS: The insoluble indigestible fraction (IIF) was higher than the soluble counterpart (soluble indigestible fraction, SIF). The SIF value was highest in black beans, while no difference was observed between chickpeas and lentils. Black beans and lentils had higher polyphenols content than chickpeas. The IF of black beans exhibited the lowest and chickpeas the highest associated polyphenols content. Condensed tannins were retained to some extent in the IF that exhibited significant antioxidant capacity. The total IF of the three pulses produced short chain fatty acids (SCFA) after 24 h of in vitro fermentation by human colonic microflora. IF from black bean and lentil were best substrates for the fermentative production of butyric acid. CONCLUSIONS: It is concluded that the IF of pulses might be an important source of bioactive compounds. Copyright © 2010 Society of Chemical Industry