Comparative Study of Chemical, Biochemical Characteristic and ATR-FTIR Analysis of Seeds, Oil and Flour of the Edible Fedora Cultivar Hemp (Cannabis sativa L.).

A series of chemical and biochemical parameters of edible hemp resources (seeds, oil, and flour) from the monoecious EU registered hemp genotype Fedora, was determined, including fatty acid profile, phytosterol composition, total phenolics, antioxidant activity, macro- and micro-elements. The fatty acid ω-3/ω-6 approached the nutritionally optimal 3/1 ratio. ß-sitosterol and other phytosterols sterols dominated the unsaponifiable fraction. Hemp seeds, flour, and oil contained 767 ± 41, 744 ± 29, and 21 ± 5 mg GAE kg-1 total polyphenols, respectively. The antioxidant potential of Fedora flour and seeds, evaluated through the DPPH (2,2-Diphenyl-1-picrylhydrazyl) assay, was higher than that of oil. K and Mg were the most abundant macro-elements, particularly in flour, while the concentration of trace elements was Fe > Cu > Ni > Mn. The presence of an array of bioactive compound candidate Fedora products as health-promoting food matrices. The ATR-FTIR spectra of hemp-derived products indicated the proximate composition of macro-nutrients.

In Vivo Release Kinetics and Antibacterial Activity of Novel Polyphenols-Enriched Chewing Gums.

Chewing gums may be particularly effective means for delivering and maintaining bioactive molecules, included in the gum formulation, able to have an anti-cariogenic effect. The purposes of this study were: to develop novel chewing gums containing quercetin (Qt); to evaluate their release using in vivo trial; finally, to test their in vivo antibacterial effect against oral Streptococcus mutans strains. A preliminary study was performed to produce new gums, enriched with the polyphenol quercetin. Then, a first in vivo experimental study was assessed to test the percentages of Qt released in the saliva of young volunteers. Moreover, a second clinical trial was performed to analyze the antibacterial capability of these enriched chewing gums against S. mutans strains after 14 days of daily consumption. The release analysis showed that a more effective release of Qt occurs in the first minutes of chewing, and it does not change saliva pH values. Moreover, Qt included in gums demonstrates an effective antibacterial activity, showing a reduction of the concentration of S. mutans strains in saliva samples, especially after 7 days. Qt included in experimental chewing gums could be efficiently released into the oral cavity and could promote an effective anti-caries concentration in volunteer's saliva, without changing salivary pH values.

Use of solid-phase microextraction coupled to gas chromatography-mass spectrometry for determination of urinary volatile organic compounds in autistic children compared with healthy controls.

Autism spectrum disorders (ASDs) are a group of neurodevelopmental disorders which have a severe life-long effect on behavior and social functioning, and which are associated with metabolic abnormalities. Their diagnosis is on the basis of behavioral and developmental signs usually detected before three years of age, and there is no reliable biological marker. The objective of this study was to establish the volatile urinary metabolomic profiles of 24 autistic children and 21 healthy children (control group) to investigate volatile organic compounds (VOCs) as potential biomarkers for ASDs. Solid-phase microextraction (SPME) using DVB/CAR/PDMS sorbent coupled with gas chromatography-mass spectrometry was used to obtain the metabolomic information patterns. Urine samples were analyzed under both acid and alkaline pH, to profile a range of urinary components with different physicochemical properties. Multivariate statistics techniques were applied to bioanalytical data to visualize clusters of cases and to detect the VOCs able to differentiate autistic patients from healthy children. In particular, orthogonal projections to latent structures discriminant analysis (OPLS-DA) achieved very good separation between autistic and control groups under both acidic and alkaline pH, identifying discriminating metabolites. Among these, 3-methyl-cyclopentanone, 3-methyl-butanal, 2-methyl-butanal, and hexane under acid conditions, and 2-methyl-pyrazine, 2,3-dimethyl-pyrazine, and isoxazolo under alkaline pH had statistically higher levels in urine samples from autistic children than from the control group. Further investigation with a higher number of patients should be performed to outline the metabolic origins of these variables, define a possible association with ASDs, and verify the usefulness of these variables for early-stage diagnosis.

[Unhealthy food choices of adolescent schoolchildren in the Campania region (Italy)]. / Scelte alimentari errate tra scolari adolescenti della regione Campania.

A questionnaire study to evaluate the eating habits of a sample of adolescents was conducted in 2009-2010 in Campania (Italy). The study involved 1,066 secondary school students aged 15 to 16 years. Fifty-two percent of participants were male and 96% lived in an urban area. Questionnaires were self-administered and anonymous. Results show an association between the consumption of different types of alcoholic beverages. Adolescents who drink carbonated beverages as a mid-morning snack tend to do the same during the mid-afternoon snack. Moreover, drinking carbonated beverages during morning and afternoon snacks was associated with the use of vending machines. Twenty-nine percent of participating students reported eating red meat more than four times per week.

Computerized sperm motility analysis in toxicity bioassays: a new approach to pore water quality assessment.

The aim of this study was to test the sensitivity of computerized sperm motility analysis in the sea urchin Paracentrotus lividus as the endpoint in toxicity bioassays. The tested matrices were pore water samples collected in an agriculture-impacted Mediterranean lagoon, Lake Varano (Italy). Two standardized bioassays were also conducted as controls, the P. lividus spermiotoxicity test and the Vibrio fischeri (Microtox®) test. VCL (curvilinear velocity), VSL (straight line velocity), VAP (average path velocity), and the percentage of rapid spermatozoa recorded by the Sperm Class Analyzer® system showed high sensitivity and discrimination ability, to a degree comparable with the larval development endpoint of the spermiotoxicity test. The test evaluated in this study requires small volumes of matrices, involves minimal sample manipulation, and can easily be extended to many other bioindicator species. It may therefore be considered a promising "quick response tool" following hazardous events that may adversely affect an aquatic ecosystem.

Active Edible Polysaccharide-Based Coating for Preservation of Fresh Figs (Ficus carica L.).

Fresh figs are very sensitive to microbial spoilage, even in cold storage conditions. Thus, fresh figs are high perishable products during postharvest with microbiological decay that induces an unpleasant taste and smell due to rot, and suitable conservation methods must therefore be applied. The fruit usually is consumed fresh locally, dried, or preserved longer term in other transformed forms. A sustainable approach to extend the shelf-life of figs can be constituted by application of an edible coating able to maintain the quality of the fruit during storage. A comparison between fresh figs in a commercial preservation system, with the figs preserved in an edible coating, and an active edible coating to preserve their quality characteristics was carried out. The coating efficacy was enhanced with the addition of pomegranate peel extract at two different concentrations. The inclusion of a component with high antioxidant activity in an edible coating proved to be an excellent method for preserving the quality of this highly perishable fruit. The application of natural products, obtained from renewable sources, represents a simple and economic strategy, but also a tool capable of preserving the quality of the fruit during the postharvest storage, which is often consumed in production areas due to shelf-life problems.

Rapid Evaluation Methods for Quality of Trout (Oncorhynchus mykiss) Fresh Fillet Preserved in an Active Edible Coating.

In this study different methods were used to evaluate the effectiveness of a carrageenan coating and carrageenan coating incorporating lemon essential oil (ELO) in preserving the physicochemical and olfactory characteristics of trout fillets stored at 4 °C up to 12 days. The fillet morphological structure was analyzed by histological and immunological methods; lipid peroxidation was performed with the peroxide and thiobarbituric acid reactive substances (TBARS) tests. At the same time, two less time-consuming methods, such as Attenuated Total Reflectance-Fourier Transformed Infrared (ATR-FTIR) spectroscopy and the electronic nose, were used. Uncoated trout fillets (UTF) showed a less compact tissue structure than carrageenan-coated threads (CTF) and coated fillets of carrageenan (active) ELO (ACTF), probably due to the degradation of collagen, as indicated by optical microscopy and ATR-FTIR. UTF showed greater lipid oxidation compared to CTF and ACTF, as indicated by the peroxide and TBARS tests and ATR-FTIR spectroscopy. The carrageenan coating containing ELO preserved the olfactory characteristics of the trout fillets better than the carrageenan coating alone, as indicated by the electronic nose analysis. This study confirms that both carrageenan and ELO containing carrageenan coatings slow down the decay of the physicochemical and olfactory characteristics of fresh trout fillets stored at 4 °C, although the latter is more effective.

Pectin-honey coating as novel dehydrating bioactive agent for cut fruit: Enhancement of the functional properties of coated dried fruits.

In this paper, a novel and sustainable process for the fruit dehydration was described. Specifically, edible coatings based on pectin and honey were prepared and used as dehydrating and antimicrobial agents of cut fruit samples, in this way promoting the fruit preservation from irreversible deteriorative processes. Pectin-honey coating was tested on apple, cantaloupe melon, mango and pineapple. The analysis were performed also on uncoated dehydrated fruits (control). The coated fruit evidenced enhanced dehydration percentage, enriched polyphenol and vitamin C contents, improved antioxidant activity and volatile molecules profile. Moreover, the antimicrobial activity against Pseudomonas and Escherichia coli was assessed. Finally, morphological analysis performed on fruit fractured surface, highlighted the formation of a non-sticky and homogeneous thin layer. These outcomes suggested that the novel fruit dehydration process, performed by using pectin-honey coating, was able to both preserve the safety and quality of dehydrated fruits, and enhance their authenticity and naturalness.

In Vitro Antibacterial Activity of Pomegranate Juice and Peel Extracts on Cariogenic Bacteria.

AIM: To evaluate the antimicrobial activity of hydroalcoholic extracts of pomegranate (Punica granatum L.) peel and juice, against the microorganisms considered the main etiologic agents of dental caries. METHODS: The values of the minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) were determined against Streptococcus mutans Clarke ATCC® 25175™ strain and Rothia dentocariosa clinical isolate. RESULTS: Peel extracts inhibit effectively the growth and survival of S. mutans ATCC 25175 strain and R. dentocariosa clinical isolate with MIC and MBC values of 10 µg/µl and 15 µg/µl, respectively. Furthermore, the pomegranate juice extract showed high inhibitory activity against S. mutans ATCC 25175 strain with a MIC value of 25 µg/µl and a MBC value of 40 µg/µl, whereas, against R. dentocariosa, it has displayed a moderate inhibitory activity, with MIC and MBC values of 20 µg/µl and 140 µg/µl, respectively. CONCLUSIONS: In vitro microbiological tests demonstrate that the hydroalcoholic extracts of pomegranate juice and peel are able to contrast the main cariogenic bacteria involved in tooth decay. Although being preliminary data, our results suggest that pomegranate polyphenolic compounds could represent a good adjuvant for the prevention and treatment of dental caries.

Isoflavone Extracts Enhance the Effect of Epidermal Growth Factor Receptor Inhibitors in NSCLC Cell Lines.

AIM: We investigated the effects of the pharmacological inhibition in vitro of epidermal growth factor receptor (EGFR) in combination with isoflavones. MATERIALS AND METHODS: Four anticancer drugs (erlotinib, gefitinib, afatinib and AZD9291) were combined with soy and red clover isoflavone extracts and used in cellular proliferation assays. The antitumor activity of inhibitors alone and in combination with isoflavone extracts was compared on three non-small cell lung cancer (NSCLC) cell lines with affiant EGFR genotype: A549 (EGFR wt); H1795 (EGFR T790M); HCC827 (EGFR del E746-A750). RESULTS: Combined treatment with extracts significantly enhanced the antiproliferative activity of all inhibitors against these cell lines. Bioactive compounds of extracts may synergize the antitumor efficacy of the inhibitors. CONCLUSION: To date, as far as we are aware, this is the first report of the combined effect of isoflavone extracts and EGFR inhibitors on human NCSLC cell growth. Sequential treatment with these drugs combined with isoflavones may represent the basis for a new therapeutic approach.

Human semen as an early, sensitive biomarker of highly polluted living environment in healthy men: A pilot biomonitoring study on trace elements in blood and semen and their relationship with sperm quality and RedOx status.

The Campania region in Italy is facing an environmental crisis due to the illegal disposal of toxic waste. Herein, a pilot study (EcoFoodFertility initiative) was conducted to investigate the use of human semen as an early biomarker of pollution on 110 healthy males living in various areas of Campania with either high or low environmental impact. The semen from the "high impact" group showed higher zinc, copper, chromium and reduced iron levels, as well as reduced sperm motility and higher sperm DNA Fragmentation Index (DFI). Redox biomarkers (total antioxidant capacity, TAC, and glutathione, GSH) and the activity of antioxidant enzymes in semen were lower in the "high impact" group. The percentage of immotile spermatozoa showed a significant inverse correlation with TAC and GSH. Overall, several semen parameters (reduced sperm quality and antioxidant defenses, altered chemical element pattern), which were associated with residence in a high polluted environment, could be used in a further larger scale study, as early biomarkers of environmental pollution.

Content of micronutrients, mineral and trace elements in some Mediterranean spontaneous edible herbs.

BACKGROUND: The analysis of mineral elements composition was determined in three wild edible herbs (Cichorium intybus L., Sonchus asper L. and Borago officinalis) collected in seven different sampling sites which were characterized by different pollution grade. The detection of mineral elements (Ca, K, Mg and Na), micronutrients (Cu, Fe, Li, Mn and Zn) and heavy metals (As, Cd, Hg, Ni and Pb) was performed. RESULTS: The results obtained show that in most cases a direct relationship appeared between the amount of elements and the sampling sites. The highest concentrations of heavy metals were found in samples grown in polluted soils. These evaluations showed that contaminants in plants may reflect the environmental state in which they develop. CONCLUSION: The examined species are a good source of mineral elements and micronutrients, making them particularly adapt to integrate a well-balanced diet. The accumulation of heavy metals showed that contaminants in plants may reflect the environmental state in which they develop. Results showed high concentrations of heavy metals in samples taken in locations characterized by high human activity and in some samples from the local market, of which no one knows the collection area.

Release kinetics of calcium and quercetin from chewing gum as a novel antiplaque and antimicrobial device.

There is a growing interest in the use of anticariogenic and antimicrobial agents to prevent demineralization and promote remineralization of dental enamel for the prevention of oral diseases. The purpose of this study was to develop a novel chewing-system containing as Casein-Phosphopeptides-Amorphous-Calcium-Phosphate (CPP-ACP) as Quercetin (Qt) and evaluate their release using in vitro and in vivo experiments. In vitro and in vivo experimental protocols were designed to test the percentages of Ca from CPP-ACP and Qt released with time and their delivery rate from a chewing gum. The in vitro experiments were performed using a specifically designed chewing apparatus to test the release of Ca and Qt in artificial saliva in function of chewed time, while the in vivo experiments required a chew-out method with volunteers and the residual CPP-ACP and Qt present in the chewed gums was analyzed. This study demonstrated that both CPP-ACP'Ca and Qt were released during chewing although CPP-ACP'Ca was released from the chewing gum in a slower and more controlled manner than Qt. They were also both effectively retained in the oral cavity for all the investigated chewing time and according to their monitored release concentrations they were able to serve as a novel antiplaque agent. This research concludes that both drugs were efficiently released during the mastication process to fully exploit their anti-carious activity.

Cytokinome profile of patients with type 2 diabetes and/or chronic hepatitis C infection.

Both type 2 diabetes (T2D) and chronic hepatitis C (CHC) infection are associated with increased risk of developing hepatocellular carcinoma (HCC). Cytokines are known to play an important role not only in the mechanisms of insulin resistance and glucose disposal defects but also in the pathological processes occurring in the liver during viral infection. We evaluated the serum levels of many cytokines, chemokines, adipokines and growth factors in patients with type 2 diabetes, CHC, CHC-related cirrhosis, CHC and type 2 diabetes and CHC-related cirrhosis and type 2 diabetes by BioPlex assay. The obtained data evidenced that the serum levels of some proteins are significantly up-regulated in all the patients or in those with only one disease and are often higher, even if in different amounts, when both diseases are associated. In particular, our results can be useful for the clinical monitoring of patients because they give specific information in regard to the progression from CHC to LC and CHD to LCD. Moreover, some molecules have shown significant correlations with clinical/biochemical data, suggesting the possibility to define mini-panels that can be used as specific markers for the different disease staging. However, our observations demonstrate that an integrated approach is much more powerful than isolated measurements to evaluate specific stages of these two complex pathologies (type 2 diabetes and chronic CHC hepatitis) alone or when they are concomitant in a patient. In fact it has emerged as an accurate, simple, specific, noninvasive, reproducible and less expensive method that, in future, could be included in routine clinical practice to monitor the association of type 2 diabetes and/or CHC to liver cirrhosis and, possibly, to cancer, and to improve the prognosis of these diseases.

A possible predictive marker of progression for hepatocellular carcinoma.

The correlation between decreased levels of selenium and increased DNA damage and oxidative stress shows the significance of this trace element. A number of studies have provided evidence for lower serum, plasma and tissue levels of selenium in patients with various diseases and types of cancer. In this study, liver selenium concentrations were measured in tissue samples of patients with hepatocellular carcinoma (HCC) by atomic absorption spectrometry. The results showed that the selenium concentrations decreased when the malignant grade increased. Furthermore, a significant correlation was found between selenium levels and human selenium binding protein-1 (SELENBP1) down-regulation in the liver. Therefore, we suggest that the evaluation of selenium and SELENBP1 concentrations can be used for improving the prognosis of HCC.

Matrix-assisted laser desorption ionization-time of flight mass spectrometry for the discrimination of food-borne microorganisms.

A methodology based on matrix-assisted laser desorption ionization-time of flight mass spectrometry of intact bacterial cells was used for rapid discrimination of 24 bacterial species, and detailed analyses to identify Escherichia coli O157:H7 were carried out. Highly specific mass spectrometric profiles of pathogenic and nonpathogenic bacteria that are well-known major food contaminants were obtained, uploaded in a specific database, and made available on the Web. In order to standardize the analytical protocol, several experimental, sample preparation, and mass spectrometry parameters that can affect the reproducibility and accuracy of data were evaluated. Our results confirm the conclusion that this strategy is a powerful tool for rapid and accurate identification of bacterial species and that mass spectrometric methodologies could play an essential role in polyphasic approaches to the identification of pathogenic bacteria.