Induction of electrophile-responsive element (EpRE)-mediated gene expression by tomato extracts in vitro

The market for food products with additional health benefits is increasing rapidly and tools for identification of bio-functional characteristics of food items are essential. To facilitate the detection of beneficial effects of tomato on gene expression, methods to prepare tomato extracts suitable to test in the EpRE LUX assay and other cell-based reporter gene assays for health-related bioactivity mechanisms, were developed. An isoprenoid-containing chloroform extract of tomato fruit and most individual isoprenoids did not induce electrophile-responsive element (EpRE)-mediated gene expression. A semi-polar extract of tomato fruits, enzymatically hydrolysed to remove the glycosyl residues from the phenolic ingredients was able to induce EpRE-mediated luciferase expression at both mRNA and protein level, which might be partly due to the presence of quercetin, kaempferol, naringenin and naringenin chalcone. It was concluded that induction of EpRE-regulated genes, such as detoxifying phase II and antioxidant enzymes, may contribute to the beneficial health effects of tomato.     

New or lesser known cultivar selection as a tool for sensory and nutritional value enhancement of osmo-convectively dried sour cherries

Taking into consideration the economic importance of sour cherry growing in Europe as well as the arising perspective of novel sour cherry product development, an investigation outlining the processing usefulness of some promising sour cherry cultivars that could lead to a better final product quality was undertaken. Nine new or lesser popular cultivars of sour cherries were compared to ‘English Morello’ with particular consideration given to processing suitability for osmo-convective drying. The quality of raw material was characterized taking into account fruit size, soluble solids content, acidity and pro-health properties. To assess the quality trait stability of individual cultivars, standard deviations were calculated. As a measure of ascertaining the fruits' suitability for drying, the sensory properties of osmo-dried products produced by the examined cultivars were considered. The gathered data demonstrates high seasonal variations in quality traits of the most investigated cultivars, which could well restrict their processing usefulness, especially for osmo-drying, as this product was found highly sensitive to fluctuation of raw material uniformity. Among the tested cultivars ‘Nefris’ emerged the most suitable for processing purposes. Fruits of this cultivar gave high quality dried product of repeatable sensory attributes characterized by significantly better pro-health properties than that of ‘English Morello’.     

Evaluation of Fourier transform infrared (FT-IR) spectroscopy and chemometrics as a rapid approach for sub-typing Escherichia coli O157:H7 isolates

The importance of tracking outbreaks of foodborne illness and the emergence of new virulent subtypes of foodborne pathogens have created the need for rapid and reliable sub-typing methods for Escherichia coli O157:H7. Fourier transform infrared (FT-IR) spectroscopy coupled with multivariate statistical analyses was used for sub-typing 30 strains of E. coli O157:H7 that had previously been typed by multilocus variable number tandem repeat analysis (MLVA) and pulsed field gel electrophoresis (PFGE). Hierarchical cluster analysis (HCA) and canonical variate analysis (CVA) of the FT-IR spectra resulted in the clustering of the same or similar MLVA types and separation of different MLVA types of E. coli O157:H7. The developed FT-IR method showed better discriminatory power than PFGE in sub-typing E. coli O157:H7. Results also indicated the spectral relatedness between different outbreak strains. However, the grouping of some strains was not in complete agreement with the clustering based on PFGE and MLVA. Additionally, HCA of the spectra differentiated the strains into 30 sub-clusters, indicating the high specificity and suitability of the method for strain level identification. Strains were also classified (97% correct) based on the type of Shiga toxin present using CVA of the spectra. This study demonstrated that FT-IR spectroscopy is suitable for rapid (≤16 h) and economical sub-typing of E. coli O157:H7 with comparable accuracy to MLVA typing. This is the first report of using an FT-IR-based method for sub-typing E. coli O157:H7.     

Papaya: Nutritional and pharmacological characterization, and quality loss due to physiological disorders. An overview

The papaya (Carica papaya L.) is a tropical fruit that is widely cultivated and consumed, both for its agreeable flavor as well as its many pharmacological properties. This review will discuss the fruit's origin and principal growing regions in the world and will briefly explore its nutritional and pharmacological attributes. In addition, we will identify and comment on some of the most common physiological disorders that occur postharvest. Such disorders compromise the quality of the fruit, bringing financial losses to the productive sector, along with serious economic and social consequences to papaya-growing countries. Among these disorders, physiological bruising, also known as “skin freckles”, characterized by the appearance of blemishes on the fruit while still in its growth stage, is one of the main problems associated with the crop. Possible causes of and current information on bruising are dealt with in this article. Other physiological disorders of the papaya such as pulp flesh translucency, pulp softening, and hard lumps in papaya flesh are also discussed.     

Feasibility of NIR spectroscopy for non-destructive characterization of table olive traits

Near infrared spectroscopy (NIRS) was used to assess the properties of intact olives. Spectral data over a range of wavelengths from 400 to 2498 nm were recorded for 448 fruit samples collected in 2008 and 2009 from seedlings of different crosses in a table olive breeding program. Although it was confirmed that the predictive models were not transferable from 1 year to the next, accurate calibration models were obtained for chemical parameters, such as oil and moisture contents (RPD values between 4.83-4.91 and 2.70-3.01, respectively). By combining the 2008 and 2009 data sets, good calibration models were obtained for some interesting physical parameters, such as fruit weight, fruit equatorial diameter and fruit volume, with RPD values of 3.34, 3.36 and 3.25, respectively. Poorer accuracy was observed for the fruit longitudinal diameter, pulp-stone ratio, endocarp equatorial diameter and stone weight, with RPD values of 2.12, 2.09, 1.84 and 1.78, respectively. The results show that NIRS is a useful, rapid and non-destructive technique for the table olive dressing industry and genotype selection programs.     

Use of Fourier transform infrared spectroscopy for rapid comparative analysis of Bacillus and Micrococcus isolates

The identification of foodborne microorganisms and their endospores in food products are important for food safety. The present work compares Bacillus (Bacillus licheniformis, Bacillus circulans and Bacillus subtilis) and Micrococcus (Micrococcus luteus) species with Fourier transform infrared (FTIR) spectroscopy. Our results show that there are several characteristic peaks belonging to both the Micrococcus and Bacillus species which can be used for the identification of these foodborne bacteria and their endospores. For Micrococcus species, a new band was observed at 1338 cm⁻¹ which may be due to acetate oxidation via the carboxylic acid cycle. The bands at 1313 cm⁻¹ and 1256 cm⁻¹ can be explained by an exopolymer formation and the other bands at 1074 cm⁻¹ and 550 cm⁻¹, may be due to the glycogen-like storage material in Micrococcus spp. There are also characteristic peaks at 993 cm⁻¹ and 801 cm⁻¹ for these bacterial species. Different Bacillus species also showed characteristic peaks at 1000–500 cm⁻¹ region. Dipicolinic acid (DPA) bands at ∼728 cm⁻¹ and ∼703 cm⁻¹ seen only in B. circulans were the marker of an endospore formation.     

Kinetics of resuscitation and growth of L. monocytogenes as a tool to select appropriate enrichment conditions as a prior step to rapid detection methods

Rapid methods still rely on a prior (shortened) enrichment step before application. Quantitative information is a prerequisite for understanding the resuscitation kinetics of the growth during the enrichment step. In this study various basal and newly introduced selective enrichment broths were evaluated. First, growth parameters (λ, μ_max) of both healthy and sub-lethally injured cells were determined. Next, a selection of enrichment broths was compared for their capacity to support detection within 24 h of low numbers of Listeria monocytogenes in artificially and naturally contaminated food samples. Detection was performed either by phage protein-based capture (Listeria Capture kit, Profos, Regensburg, Germany) combined with plating on chromogenic medium or by fluorescence in situ hybridization (FISH) using the VIT-Listeria kit (Vermicon, Munich, Germany). Kinetics of resuscitation and growth of L. monocytogenes in various enrichment broths showed that for detection of low numbers of sub-lethally injured L. monocytogenes cells at least an overnight enrichment was needed. A selective enrichment broth was needed to enable proliferation of L. monocytogenes within the indigenous bacterial flora present in foods. However, combination of an appropriate enrichment condition with advanced detection techniques may enable a 24 h detection of L. monocytogenes.     

Quantification of aspartame in commercial sweeteners by FT-Raman spectroscopy

Efficient methods are proposed herein for the quantification of aspartame in commercial sweeteners. These methods are based on a treatment of Raman data with partial least squares (PLS), principal component regression (PCR) and counter-propagation artificial neural networks (CP-ANN) methods. For the three chemometric techniques used, the relative standard errors of prediction (RSEP) calculated for calibration and validation data sets were on the order of 1.8–2.2%. Four commercial preparations containing between 17% and 36% of aspartame by weight were evaluated by applying the developed models. Concentrations found from the Raman data analysis agree perfectly with the results of the UV–Vis reference analysis, with the recoveries in the 98.7–100.8%, 98.6–101.1% and 97.8–102.2% ranges for the PLS, PCR and CP-ANN models, respectively. The proposed procedures can be used for routine quality control during the production of commercial aspartame sweeteners.     

Ustilago maydis killer toxin as a new tool for the biocontrol of the wine spoilage yeast Brettanomyces bruxellensis

Brettanomyces bruxellensis is one of the most damaging species for wine quality, and tools for controlling its growth are limited. In this study, thirty-nine strains belonging to Saccharomyces cerevisiae and B. bruxellensis have been isolated from wineries, identified and then tested against a panel of thirty-nine killer yeasts. Here, for the first time, the killer activity of Ustilago maydis is proven to be effective against B. bruxellensis. Mixed cultures in winemaking conditions show that U. maydis CYC 1410 has the ability to inhibit B. bruxellensis, while S. cerevisiae is fully resistant to its killer activity, indicating that it could be used in wine fermentation to avoid the development of B. bruxellensis without undesirable effects on the fermentative yeast. The characterization of the dsRNAs isolated and purified from U. maydis CYC 1410 indicated that this strain produces a KP6-related toxin. Killer toxin extracts were active against B. bruxellensis at pH values between 3.0 and 4.5 and temperatures comprised between 15 °C and 25 °C, confirming their biocontrol activity in winemaking and wine aging conditions. Furthermore, small amounts (100 AU/ml) of killer toxin extracts from U. maydis significantly reduced the amount of 4-ethylphenol produced by B. bruxellensis, indicating that in addition to the growth inhibition observed for high killer toxin concentrations (ranging from 400 to 2000 AU/ml), small amounts of the toxin are able to reduce the production of volatile phenols responsible for the aroma defects in wines caused by B. bruxellensis.     

HPLC quantification of major active components from 11 different saffron (Crocus sativus L.) sources

Eleven certified saffron samples (Crocus sativus L.), one each from Azerbaijan, China, Greece, France, India, Iran, Italy, New Zealand, Spain, Turkey and the Sigma Chemical Company, were analyzed by using an HPLC photodiode array detection method. This analysis quantified the 10 major saffron compounds in each sample and their concentration was analyzed at three different wavelengths. Results indicated that the Greek, Indian, New Zealand, and Spanish saffron extracts possessed the highest concentrations of water-soluble glycosidic carotenoids (?8.0%) suggesting that they could be a good source of this type of metabolites for further biological evaluation.     

Identification of seven species of the Lactobacillus acidophilus group by FT-IR spectroscopy

Fourier transform infrared (FT-IR) spectra of 102 strains of the seven species of the Lactobacillus acidophilus group were collected and investigated for their potential use in classification and identification on species level. The database built contains more than 370 spectra. Various procedures of pre-processing and classification methods have been compared with respect to their predictive ability. The most encouraging results were achieved with linear discriminant analysis (LDA) of the absorbance values of normalized spectra at selected wavenumbers. The rate of correct species assignment in cross-validation (Jackknife procedure with one spectrum left out for model building) were 95%, 95%, 69%, 100%, 88%, 100%, and 91% for L. acidophilus, L. amylovorus, L. crispatus, L. gallinarum, L. gasseri, L. helveticus, and L. johnsonii, respectively. Very distinct grouping was found for L. gallinarum and L. helveticus, the most difficult differentiation in LDA was between the pairs L. crispatus/L. amylovorus and L. gasseri/L. johnsonii.     

Efficacy of silkworm (Bombyx mori L.) chrysalis oil as a lipid source in adult Wistar rats

The effects of silkworm chrysalis oil, rich in n-3 α-linolenic acid (ALA), on lipid metabolism in Wistar rats were investigated. The rats were fed diets containing 7% soybean oil (control), silkworm chrysalis oil (SWO), or fish oil (FO) for 8 weeks. Plasma triglyceride and glucose levels were significantly lower in the SWO group after 8 weeks compared to the control and FO groups. The total cholesterol and blood urea nitrogen levels were higher in the control group than in the SWO and FO groups at 8 weeks post-consumption. However, aspartate amino transferase and alanine amino transferase levels were not significantly different among all groups. A higher arachidonic acid (AA) content was detected in the control group, while lower AA levels were observed with the increase in EPA and DHA in the SWO and FO groups. These results suggest that n-3 α-linolenic acid-rich silkworm chrysalis oil can improve hyperlipidaemia and hyperglycaemia.     

Purification, antitumour and immunomodulatory activity of water-extractable and alkali-extractable polysaccharides from Solanum nigrum L.

The water-extractable and the alkali-extractable polysaccharides from Solanum nigrum L. (named SNLWP and SNLAP, respectively) and their four polysaccharide sub-fractions (SNLWP-1, SNLWP-2, SNLAP-1 and SNLAP-2), were isolated and purified by ethanol precipitation, dialysis, anion-exchange and gel filtration chromatography. Antitumour and immunomodulatory activity of the polysaccharides was evaluated by determining the survival time, the ascites volume, the weight of immune organ and the level of cytokine (IL-2, IL-10 and IFN-γ) of H₂₂-bearing mice, respectively. The results showed that SNLWP-1, SNLAP-1 and SNLAP-2 had significant antitumour and immunomodulatory activity, whereas SNLWP-2 hardly demonstrated the activity. SNLWP-1 contained galactose and arabinose as main sugar components, and both SNLAP-1 and SNLAP-2 were rich in xylose, galactose and arabinose. SNLWP-2 was rich in glucose. In conclusion, SNLWP-1, SNLAP-1 and SNLAP-2 have potent antitumour activity which may be associated with their potent immunostimulating effect and monosaccharide composition.     

Comparison of the cell wall composition for flesh and skin from five different plums

Cell walls were isolated from flesh and skin of five plum varieties corresponding to three species (Prunus domestica L., Prunus salicina Lindl. and Prunus insititia Lindl.) using the alcohol-insoluble solids (AIS) procedure. Yields varied from 83 to 114 g kg⁻¹ dry weight in the flesh and from 192 to 361 g kg⁻¹ dry weight in the skins. Their main sugars were uronic acid (224–322 mg g⁻¹ AIS), cellulosic glucose (139–170 mg g⁻¹ AIS), galactose and arabinose. Galactose and arabinose ratio were variable between the varieties. The degrees of methylation were high (62–84).     

A rapid microwave-assisted derivatization process for the determination of phenolic acids in brewer’s spent grains

A simple and fast method for the TMS derivatization of phenolic acids in a closed vial using microwave irradiation followed by GC/MS analysis is proposed. A full factorial design was used to investigate the effects of two independent variables, namely, time and power of microwave irradiation, on the yield of silylation reaction. The optimal conditions were tested against the classical heating derivatization procedure. No significant differences were found between the classical heating and microwave-assisted derivatization. Chromatographic separation of the nine phenolic acids examined was achieved in 16 min. The mass spectral fragmentation patterns of the derivatives obtained by the proposed method were identical to those from the classical heating. Four different batches of brewer’s spent grain were extracted and analyzed for the total phenolic acid content. Significant differences between the batches of spent grains were found for all analytes. The total phenolic acid content varied between 2688 and 4884 μg/g.     

A rapid screening for adulterants in olive oil using DNA barcodes

A distinctive methodology is developed to trace out the mixing into olive oil, which is marketed every year with 20% or more fraudulent oils. Such adulteration has been difficult to differentiate using fatty acid analysis and other available current techniques, as chemically fatty acids are same regardless of their source. The total genomic DNA isolated from olive oil, contaminated with canola and sunflower was analysed for single nucleotide polymorphism (SNP) variation in noncoding spacer region between psbA-trnH and partial coding region of matK of plastid genome. These DNA regions were amplified by PCR using specific primers and resulting DNA sequences were matched to the predetermined consensus DNA barcode sequences of canola and sunflower for discerning the contaminations in olive oil samples. The matching of an adulterant DNA sequence with their respective DNA barcode revealed the mixing of canola and sunflower oil into olive is simpler way and the combined approach of molecular biology and bioinformatics technology can be used as an inexpensive method for ensuring the purity of olive. This plastid based molecular DNA technology can be used for rapid detection of adulteration easily up to 5% in olive oil.     

Cooking quality of faba bean after storage at high temperature and the role of lignins and other phenolics in bean hardening

Selected physical and chemical characteristics of faba beans (Vicia faba L.) cv. Fiesta were studied after 12 months storage at 5, 15, 25, 37, 45 or 50 °C (±2 °C) in relation to the hard-to-cook phenomenon. In comparison with control (seeds stored at 5 °C), seeds stored at 15 and 25 °C demonstrated non-significant (p?0.05) changes in most of the physical and chemical characteristics including hydration and swelling coefficients, acid detergent fibre, lignin and tannin contents, whereas seeds stored at ?37 °C demonstrated significant changes (p?0.05). Solutes and electrolytes leaching after 18 h soaking substantially increased with increased temperature. Faba bean hardness tested by the hard-to-cook test also increased substantially with increased storage temperature. After 8 h soaking followed by 2 h cooking, the puncture force required for seeds stored at 5 °C was 3.3 N seed⁻¹ whereas seeds stored at 50 °C required a much higher puncture force of 15.2 N seed⁻¹. There was a high negative correlation (r²=0.98) between storage temperature and cooking ability of faba bean. Substantial increases in acid detergent fibre and lignin contents occurred with increased storage temperatures. There was a three-fold increase in lignin content of faba bean stored at 50 °C compared to those stored at 5 °C and it was correlated with bean hardness (r²=0.98). Storage at high temperatures for 12 months led to a substantial reduction in total free phenolics especially in the testa and there was a greater reduction with increasing storage temperature. Reduction in free phenolics was negatively correlated (r²=0.75) with bean hardness.     

Rapid primary structure determination of myoglobins by a complementary approach based on mass spectrometry and Edman degradation

The primary structure of two myoglobins (Mbs), isolated from the heart and muscle of Equus asinus L. and Struthio camelus L., respectively, was determined using a combined approach based on Edman degradation and mass spectrometry. The strategy allowed the determination of donkey Mb sequence, which was found to be identical to the horse Mb, as also confirmed by ESI/Q-TOF mass spectrometry. Indeed, donkey Mb accurate molecular mass (16951.50 Da) was in good agreement with the molecular mass of horse Mb (16951.48 Da). A similar strategy was also applied for revisiting the primary structure of ostrich Mb, revealing the presence of two amino acid substitutions (i.e. Asp53Glu and Asp60Glu), with respect to the previously reported sequence ( Enoki, Ohga, Ishidate, & Morimoto, 2008). The proposed approach represents a rapid and reliable tool for determining/revisiting the primary structures of the highly conserved myoglobins.     

HPLC profiling of phenolics in diverse potato genotypes

Potatoes from over fifty genotypes representing cultivars, breeding lines, primitive germplasm and wild species were analysed for phenolic content and hydrophilic antioxidant capacity. Genotypes with markedly higher amounts than the most commonly consumed potatoes were identified. Chlorogenic acid was the most abundant phenolic and ranged from 22 to 473 mg/100 g dry weight. Rutin and kaempferol-3-rutinose were the most abundant flavonols. Total phenolics ranged from 1.8 to 11 mg/g DW and antioxidant capacity ranged from 27 to 219 μmol TE/g DW. Total phenolics and antioxidants in these high-phytonutrient potatoes compared favourably to 15 other analysed vegetables. With the high per capita consumption of potatoes, widespread adoption of high-phytonutrient cultivars could significantly increase dietary intake of phytonutrients.