Utilization of the genetic resources of wild species to create a nontransgenic high flavonoid tomato

Flavonoids represent a large and important group of plant natural products that are ubiquitous in the plant kingdom. Epidemiological studies have shown the health benefits of a diet high in flavonoids. However, the dietary intake of flavonoids in most western populations is limited, creating a need to find alternative food sources for these polyphenolic secondary metabolites. The domestication of many of our cultivated food crops has resulted in alterations in the biosynthetic pathways of many essential micronutrients and vitamins through inadvertent counterselection against nutritional traits in favor of agronomic ones. Flavonoids are nearly absent from fruits of cultivated tomato (Lycopersicon esculentum Mill.), a major vegetable in human diets. Previous attempts to restore the flavonoid pathway in tomato fruits have been limited to transgenic strategies, suggesting that the problem was intractable through traditional methods. Here, we describe for the first time a nontransgenic metabolic engineering approach to developing a high flavonoid tomato using a wild tomato species (Lycopersicon pennelliiv. puberulum) and demonstrate the opportunities for restoring functional pathways using the genetic resources of wild species, resulting in production of healthier foods.     

Derivative potentiometric stripping analysis (dPSA) used for the determination of cadmium,copper, lead,and zinc in Sicilian olive oils

Derivative potentiometric stripping analysis (dPSA) was utilized to evaluate the Cd(II), Cu(II), Pb(II), and Zn(II) content in olive oil samples produced in Sicily in the crop year 2000-2001. The repeatability of the method was attested at 86.36% for cadmium, at 94.94% for copper, at 99.00% for lead, and at 98.92% for zinc. Recovery tests were carried out, both on cleanup procedures and on extraction steps, on olive oil spiked at different levels; obtained recoveries were 84.52 +/- 9.86 for cadmium, 97.34 +/- 2.72 for copper, 100.68 +/- 0.67 for lead and 83.35 +/- 1.72 for zinc. Theoretic detection limits were 1.2 ng g(-1) for Cd, 3.6 ng g(-1) for Cu, 5.9 ng g(-1) for Pb, and 14.3 ng g(-1) for Zn. Found concentrations range were 15.94-58.51 ng g(-1) for Cu, 32.64-156.48 ng g(-1) for Pb, and 157.00-385.22 ng g(-1) for Zn. Copper, lead, and zinc were found in all samples. The main advantage of this determination consists of a not too aggressive metals extraction procedure using hydrochloric acid, which avoids losses of elements typical of sample calcinations methods.     

Chemical characterization and antioxidant properties of coffee melanoidins

Melanoidins, the brown polymers formed through Maillard reaction during coffee roasting, constitute up to 25% of the coffee beverages' dry matter. In this study chemical characterization of melanoidins obtained from light-, medium-, and dark-roasted coffee beans, manufactured from the same starting material, was performed. Melanoidins were separated by gel filtration chromatography and studied by MALDI-TOF mass spectrometry. Results showed that the amount of melanoidins present in the brews increased as the intensity of the thermal treatment increased, while their molecular weight decreased. The antioxidant activity of melanoidins isolated from the different brews was studied by using different methodologies. Melanoidins antiradical activity determined by ABTS(*)(+) and DMPD(*)(+) assays decreased as the intensity of roasting increased, but the ability to prevent linoleic acid peroxidation was higher in the dark-roasted samples. Data suggest that melanoidins must be carefully considered when the relevance of coffee intake in human health is studied.     

Model studies on the photosensitized isomerization of bixin

The photosensitized isomerization reaction of the natural cis carotenoid bixin (methyl hydrogen 9'-cis-6, 6'-diapocarotene-6, 6'-dioate) with rose bengal or methylene blue as the sensitizer in acetonitrile/methanol (1:1) solution was studied using UV-vis spectroscopy, high-performance liquid chromatography (HPLC), and time-resolved spectroscopic techniques, such as laser-flash photolysis and singlet oxygen phosphorescence detection. In both N(2)- and air-saturated solutions, the main product formed was all-trans-bixin. The observed isomerization rate constants, k(obs), decreased in the presence of air or with increase in the bixin concentration, suggesting the participation of the excited triplet state of bixin, (3)Bix, as precursor of the cis--> trans process. On the other hand, bixin solutions in the absence of sensitizer and/or light did not degrade, indicating that the ground state of bixin is stable to thermal isomerization at room temperature. Time-resolved spectroscopic experiments confirmed the formation of the excited triplet state of bixin and its deactivation by ground state bixin and molecular oxygen quenching processes. The primary isomerization products only degraded in the presence of air and under prolonged illumination conditions, probably due to the formation of oxidation products by reaction with singlet molecular oxygen. An energy-transfer mechanism was used to explain the observed results for the bixin transformations, and the consequences for food color are discussed.     

Classification of cereal flours by chemometric analysis of MIR spectra

Different kinds of cereal flours submitted to various technological treatments were classified on the basis of their mid-infrared spectra by pattern recognition techniques. Classification in the wavelet domain was achieved by using the wavelet packet transform for efficient pattern recognition (WPTER) algorithm, which allowed singling out the most discriminant spectral regions. Principal component analysis (PCA) on the selected features showed an effective clustering of the analyzed flours. Satisfactory classification models were obtained both on training and test samples. Furthermore, mixtures of varying composition of the studied flours were distributed in the PCA space according to their composition.     

Optothermistor as a breakthrough in the quantification of lycopene content of thermally processed tomato-based foods: verification versus absorption spectrophotometry and high-performance liquid chromatography

This study reports on the first use of the "optothermistor" as a novel, precise, fast, and low-cost detector of lycopene in a wide range of commercially available processed-tomato products. The quantitative performance of the new device was evaluated by comparing data obtained to that acquired by conventional methods, namely, absorption spectrophotometry and high-performance liquid chromatography (HPLC); the linear correlation was high (R = 0.98). The variation of data obtained with the optothermistor in a series of consecutive measurements performed with the same loading of the sample was better than 1%. However, the repeatability (RSD 0.5-9.0%, n = 3-5) achieved with the optothermistor by independent analyses (multiple loading) is comparable to that of HPLC and spectrophotometry. Results of the studies performed on the 19 products derived from tomatoes demonstrated that the optothermistor is suitable for selective, accurate, precise, and simple determination of lycopene (range = 7-75 mg/100 g of product weight) without the need for a sample pretreatment step. The estimated sensitivity of the present optothermistor is 2 mg of lycopene/100 g of product.     

The elusiveness of coffee aroma: new insights from a non-empirical approach

Aroma is central to a pleasurable eating/drinking experience but is one of the most labile components of food. Coffee is an outstanding example. Attempts to avoid or control aroma degradation are often frustrated by ignorance of the microscopic mechanisms that are responsible for it. One of the processes most frequently invoked is radical formation, yet the identity of the radicals and their involvement in aroma degradation are poorly understood at the molecular level. Here a step forward in the fundamental understanding of this complex problem is taken by identifying the most relevant radicals and their products using first-principles calculations. Over 100 radicals originating from key aroma compounds found in coffee and other foods have been studied and classified according to an unambiguous criterion: their thermodynamic stability relative to common radical sources. This classification scheme predicts that most aroma molecules are resistant to both peroxidation and attack from phenolic antioxidants but are unstable with respect to radicals such as .OH. Dimers--generated from radical reactions--were also considered, and the most volatile species, which may further contribute to coffee aroma degradation, were focused on. Those--which are very few indeed--that have this potential have been identified.     

Determination of selenium in nuts by cathodic stripping potentiometry (CSP)

The aim of this work was to determine the selenium content in nut samples by cathodic stripping potentiometry. Dry-powdered nuts were digested by HNO(3) and dissolved with concentrated hydrochloric acid. To avoid the interference of natural oxygen, the potentiometric determination of selenium was carried out in an electrolyte solution consisting of 2 M CaCl(2) and 4 M HCl. The analysis was executed applying an electrolysis potential of -150 mV for 60 s and a constant current of -30 microA. Under these conditions, detection limits lower than 1.0 ng g(-)(1) were obtained for selenium analysis in nuts. The relative standard deviation of these measurements (expressed as rsd %) ranged from 0.44 to 0.88% while recoveries ranged from 90.2 to 95.3%. The results obtained with the proposed method were compared with those obtained via hydride vapor generation atomic absorption spectroscopy, a common method for determining selenium. The results of the two methods agreed within 5% for almond, hazelnut, and pistachio samples. The mean concentrations of selenium determined in Sicilian samples of almond, hazelnut, and pistachio were 531 +/- 1, 865 +/- 1, and 893 +/- 4 microg/kg, respectively.     

Interactions of prion proteins with soil

Prions, are proteinaceous particles recognized as the agents of a class of neurodegenerative disorders, called transmissible spongiform encephalopathies (TSE), or prion diseases. Epidemiological data suggest that TSE-contaminated environments may serve as source of infectivity, but there is no information about adsorption of prions onto soil. We carried out experiments by mixing, healthy, or scrapie-infected hamster brains homogenates with three types of soil suspended in different buffers: (i) two saline buffers, i.e., phosphate buffer solution (PBS) and CaCl₂ solution; (ii) a mix of nondenaturing detergents, i.e., Triton X-100 and sodium deoxycholate (DOC) solution; (iii) a non-ionic detergent, i.e., lauryl maltoside; (iv) two anionic detergents, i.e., Sarkosyl or sodium dodecyl sulphate (SDS); and (v) a chaotropic agent, i.e., urea. The unbound prion proteins were detected in the supernatants (after centrifugation of soil suspension) by Western blotting. Results clearly demonstrate that both the no infectious (PrP^C) and infectious (PrP^Sc) forms are adsorbed by all soils. Only 1% sodium dodecylsulphate (SDS) partially impeded the association of PrP^C, but not that of PrP^Sc with the sandy loam soil. Agents with different interacting properties towards hydrophilic and/or hydrophobic domains failed to extract PrP^Sc from sediments of soil-brain homogenate mixtures. The strong interaction of PrP^Sc with soil favors the accumulation of prions in soils, especially if amended with prion-containing organic fertilizers and/or whenever TSE-affected animal carcasses, placenta, and excreta in general are buried or laid at the soil surface.     

Isolation and biochemical characterization of a basic myrosinase from ripe Crambe abyssinica seeds,highly specific for epi-progoitrin

On the basis of previous studies on the mechanism-based inhibition, activation, and active site structure of myrosinase(s) isolated from Sinapis alba and other cruciferous seeds, crambe myrosinase shows uncommon properties and behavior. For this reason homogeneous crambe myrosinase was isolated and investigated to establish the most important physicochemical features, including kinetic properties determined with the epimers progoitrin (R) and epi-progoitrin (S) as substrates, with and without ascorbate as an activator. The results of this study demonstrate that crambe myrosinase is highly specific for epi-progoitrin due to a better stabilization of the enzyme-substrate complex. This stabilization is caused by additional hydrogen bonding that only epi-progoitrin can set up between its hydroxyl group and a suitable residue in the hydrophobic pocket where the "docking" of the glucosinolates side chain takes place.