Recognizing Age at Slaughter of Cattle from Beef Samples Using GC/MS–SPME Chromatographic Method

Beef from cattle less than 30 months old at the point of slaughter are generally sold in meat markets. Though the age of the animal can generally be estimated by dentition, this method cannot be applied to a carcass or a piece of meat from which the teeth have been removed. Since gas chromatography of volatile organic compounds has been used to classify biological samples, this technique was used for classifying meat obtained from cattle aged over 30 months (OTM) at point of slaughter. Volatile organic compounds of fresh meat (M. longissimus dorsi) samples obtained from cattle of different age and dentition (two, four, and six teeth) at point of slaughter were analyzed using gas chromatography/mass spectrum (GC/MS). Six volatile organic compounds were selected (hexanoic acid, benzoic acid, octanoid acid, tridecane, tetradecane, and 4-formylbenzoic acid) and confirmed by using chemical standards and Kovat’s indexes. The chromatogram data were used to train two classifiers, one based on the partial least square (PLS) method and another based on support vector machine (SVM), but only SVM showed high accuracy. Data exploration based on the principal component analysis allowed for comprehension of the data distribution, proposing a new version of PLS in a parallel configuration. This modified version of the PLS algorithm showed better accuracy than the SVM classifier. In conclusion, volatile organic compounds analyzed by GC/MS–solid-phase microextraction can be used to recognize OTM beef.     

Dehydrogenase Enzymes Associated to Glycolysis in Beef Carcasses Stored at 0 °C

After the death of an animal, cell metabolism is controlled locally. The post-mortem oxygen depletion increases the glycolytic activity and lactate production. However, many mechanisms of post-mortem metabolic regulation have not been fully investigated in beef carcasses. In this work, we studied the post-mortem glycolytic behavior (including lactate dehydrogenase) and three dehydrogenase associated to glycolysis (glycerophosphate dehydrogenase, glucose 6-phosphate dehydrogenase, and glycerol dehydrogenase) by using cytochemistry techniques in three fast-twitch muscles (M. longissimus dorsi, M. semimembranosus, and M. cutaneus trunci) of carcasses stored at 0 °C. Our results indicate that glycolysis depends on the type of muscle. The post-mortem glycolytic flux and lactate dehydrogenase activity of M. cutaneus trunci was the lowest of the three muscles studied. Of the other dehydrogenases analyzed, only glycerophosphate and glycerol dehydrogenase showed clear cytochemical reaction. Glucose 6-phosphate dehydrogenase was not used by muscles very much. The glycerophosphate dehydrogenase was the strongest enzymatic activity correlated to the post-mortem glycolytic flux. In addition, a relationship between glycerophosphate dehydrogenase and glycerol dehydrogenase was detected by using a multiple regression model. This phenomenon was studied by using bioinformatics tools, suggesting that glycerophosphate dehydrogenase could oxidize the glycerol in bovine fast-twitch muscles.     

Angiotensin-I converting enzyme inhibitory and antioxidant activity of bioactive peptides produced by enzymatic hydrolysis of skin from grass carp (Ctenopharyngodon idella)

Grass carp skin pieces were homogenized in water and hydrolyzed by Alcalase®, collagenase, proteinase K, and/or trypsin at their optimum conditions. Samples were taken at various degrees of hydrolysis and were evaluated for antioxidant, antimicrobial, and angiotensin-converting enzyme inhibitory activities. Alcalase and collagenase completely hydrolyzed the skin with different rates, and released peptides with antioxidant and angiotensin-converting enzyme-inhibitory activity. These activities increased linearly with increasing degrees of hydrolysis. Subsequent incubation of the collagenase hydrolysates with trypsin slightly increased the antioxidant activity. Proteinase K, although only partially hydrolyzing the skin, also catalyzed the release of peptides with antioxidant and angiotensin-converting enzyme-inhibitory activities. These results show that skin by-products from grass carp can be a source of bioactive peptides produced by a one-step reaction. Such hydrolysates may be applied in food products to prolong shelf life and provide beneficial effects on blood pressure.     

Changes in lipid fractions and sensory properties of Idiazabal cheese induced by lipase addition

This work studied the addition of an adequate lipase to enhance lipolysis reactions and the development of piquant flavour and sharp odour in Idiazabal cheese, as an alternative to the use of lamb rennet paste. Cheeses were manufactured from bulk raw ewes' milk in 50 l vats with commercial bovine rennet and 80 lipase units of pregastric or 180 lipase units of fungal lipase and ripened for 180 days. A higher lipolytic activity was induced by lipase addition promoting strong changes in odour and flavour attributes. Both fungal and pregastric lipases increased the content of total free fatty acids (FFA), but the fungal lipase released mainly medium- and long-chain FFA. In contrast, the pregastric lipase preferably released short-chain FFA. Diglyceride (DG) content was considerably higher in cheeses made with added pregastric lipase compared with those made with fungal lipase or with no lipase. Monoglycerides (MG) were detected only in cheeses made with either lipase added, reaching comparable concentrations after ripening for 180 days. The cheeses made with pregastric lipase had the highest scores for odour and flavour intensity, and sharp and rennet odours, desirable attributes for the Idiazabal cheese made with lamb rennet paste. None of the texture attributes were significantly influenced by the concentrations of MG and DG in the cheeses made with either lipase. Thus, the pregastric lipase was more appropriate than the fungal lipase to develop a more traditionally-flavoured Idiazabal cheese.     

Determination of a representative volume element based on the variability of mechanical properties with sample size in bread

Quantitative analysis of food structure is commonly obtained by image analysis of a small portion of the material that may not be the representative of the whole sample. In order to quantify structural parameters (air cells) of 2 types of bread (bread and bagel) the concept of representative volume element (RVE) was employed. The RVE for bread, bagel, and gelatin-gel (used as control) was obtained from the relationship between sample size and the coefficient of variation, calculated from the apparent Young's modulus measured on 25 replicates. The RVE was obtained when the coefficient of variation for different sample sizes converged to a constant value. In the 2 types of bread tested, the tendency of the coefficient of variation was to decrease as the sample size increased, while in the homogeneous gelatin-gel, it remained always constant around 2.3% to 2.4%. The RVE resulted to be cubes with sides of 45 mm for bread, 20 mm for bagels, and 10 mm for gelatin-gel (smallest sample tested). The quantitative image analysis as well as visual observation demonstrated that bread presented the largest dispersion of air-cell sizes. Moreover, both the ratio of maximum air-cell area/image area and maximum air-cell height/image height were greater for bread (values of 0.05 and 0.30, respectively) than for bagels (0.03 and 0.20, respectively). Therefore, the size and the size variation of air cells present in the structure determined the size of the RVE. It was concluded that RVE is highly dependent on the heterogeneity of the structure of the types of baked products.     

Detection and identification of tyrDC + enterococcal strains from pasteurized commercial cheeses

The aim of this study was to isolate and identify strains of Enterococcus from commercial cheeses and then analyze their abilities to produce biogenic amines. The genotypic variability, studied by randomly amplified polymorphic DNA (RAPD)-PCR, showed a high degree of homogeneity among enterococcal strains. Afterward, genotypic analysis indicated that all strains contain genes encoding a tyrosine decarboxylase. Our results indicate that a potential health risk exists if the enterococcal strains are able to survive the pasteurization of milk or appear as post-pasteurization contaminants. These results highlight the importance of the hazard analysis and critical control point (HACCP) to minimize the risk of hazards associated with post-contamination during cheese elaboration     

Diminution of quinolizidine alkaloids,oligosaccharides and phenolic compounds from two species of Lupinus and soybean seeds by the effect of Rhizopus oligosporus

Lupinus, like Glycine max (soybean), is an ancient leguminous plant. It has been used as a food by people living around the Mediterranean Sea and in the Andean Highlands. This legume contains quinolizidine alkaloids (Qas), oligosaccharides (OGS) and phenolic compounds (PC). The objective of this study was to evaluate the effect of the growth of Rhizopus oligosporus on the levels of Qas, OGS and PC from L. mutabilis, L. campestris and G. max through tempeh elaboration. The results showed that the soaking and cooking processes of legume seeds diminished the Qas content of L. mutabilis and L. campestris by 50%, and after 48 h of fermentation these compounds decreased by more than 90% in total. OGS diminished by more than 90% in the lupin seeds. The PC content of the three seed species subjected to these processes increased their absorbance value, probably due to the enzymatic action of a fungal tannase. These results suggested that the L. mutabilis, L. campestris and G. max fermentation with R. oligosporus is an efficient method for diminishing antinutritional factors and for obtaining a product with optimal nutritional value. Copyright © 2007 Society of Chemical Industry     

Adjusting Some Properties of Poly(methacrylic acid) (Nano)Composite Hydrogels by Means of Silicon-Containing Inorganic Fillers

The present work aims to show how the main properties of poly(methacrylic acid) (PMAA) hydrogels can be engineered by means of several silicon-based fillers (Laponite XLS/XLG, montmorillonite (Mt), pyrogenic silica (PS)) employed at 10 wt% concentration based on MAA. Various techniques (FT-IR, XRD, TGA, SEM, TEM, DLS, rheological measurements, UV-VIS) were used to comparatively study the effect of these fillers, in correlation with their characteristics, upon the structure and swelling, viscoelastic, and water decontamination properties of (nano)composite hydrogels. The experiments demonstrated that the nanocomposite hydrogel morphology was dictated by the way the filler particles dispersed in water. The equilibrium swelling degree (SDe) depended on both the pH of the environment and the filler nature. At pH 1.2, a slight crosslinking effect of the fillers was evidenced, increasing in the order Mt < Laponite < PS. At pH > pKa_MAA (pH 5.4; 7.4; 9.5), the Laponite/Mt-containing hydrogels displayed a higher SDe as compared to the neat one, while at pH 7.4/9.5 the PS-filled hydrogels surprisingly displayed the highest SDe. Rheological measurements on as-prepared hydrogels showed that the filler addition improved the mechanical properties. After equilibrium swelling at pH 5.4, G’ and G” depended on the filler, the Laponite-reinforced hydrogels proving to be the strongest. The (nano)composite hydrogels synthesized displayed filler-dependent absorption properties of two cationic dyes used as model water pollutants, Laponite XLS-reinforced hydrogel demonstrating both the highest absorption rate and absorption capacity. Besides wastewater purification, the (nano)composite hydrogels described here may also find applications in the pharmaceutical field as devices for the controlled release of drugs.     

STEM imaging of single Pd atoms in activated carbon fibers considered for hydrogen storage

Aberration corrected scanning transmission electron microscopy was used to demonstrate the feasibility of imaging individual Pd atoms that are highly dispersed throughout the volume of activated carbon fibers. Simultaneous acquisition of high-angle annular dark-field and bright-field images allows correlation of the location of single Pd atoms with microstructural features of the carbon host material. Sub-Ångström imaging conditions revealed that 18 wt% of the total Pd content is dispersed as single Pd atoms in three re-occurring local structural arrangements. The identified structural configurations may represent effective storage sites for molecular hydrogen through Kubas complex formation as discussed in detail in the preceding article.     

A New Smoothened Antagonist Bearing the Purine Scaffold Shows Antitumour Activity In Vitro and In Vivo

The Smoothened (SMO) receptor is the most druggable target in the Hedgehog (HH) pathway for anticancer compounds. However, SMO antagonists such as vismodegib rapidly develop drug resistance. In this study, new SMO antagonists having the versatile purine ring as a scaffold were designed, synthesised, and biologically tested to provide an insight to their mechanism of action. Compound 4s was the most active and the best inhibitor of cell growth and selectively cytotoxic to cancer cells. 4s induced cell cycle arrest, apoptosis, a reduction in colony formation and downregulation of PTCH and GLI1 expression. BODIPY-cyclopamine displacement assays confirmed 4s is a SMO antagonist. In vivo, 4s strongly inhibited tumour relapse and metastasis of melanoma cells in mice. In vitro, 4s was more efficient than vismodegib to induce apoptosis in human cancer cells and that might be attributed to its dual ability to function as a SMO antagonist and apoptosis inducer.