Chemotype and genotype combined analysis applied to tomato (Lycopersicon esculentum Mill.) analytical traceability

A large number of fresh fruits and vegetables are primary sources of antioxidants; tomato (Lycopersicon esculentum Mill.) is accepted worldwide as a significant source of antioxidant functional compounds (vitamin C, lycopene, rutin). Many cultivars and hybrids of tomato, having different chemical and nutritional characteristics, are available on the market. Tomato cultivars for industrial processing are very different, not only in fruit characteristics (size, shape), but also in lycopene and antioxidant contents. The aim of this study was the chemotyping and genotyping of the tomato varieties Heinz 3402, Leader and Perfectpeel, (1) to evaluate the genetic traceability of these varieties, and (2) to determine whether their functional antioxidants compounds are useful markers of traceability. Principal component analysis (PCA) was first applied to the Random Polymorphic DNA (RAPD) fingerprints, confirming that this approach is a powerful identification method at intra-specific level. Heinz 3402 showed the highest antioxidant activity, followed by Perfectpeel and Leader varieties. Perfectpeel showed the lower lycopene content, while Leader and Heinz 3402 showed significantly higher values (13.68 and 15.78 mg/100 g, fresh weight, respectively). The highest rutin content was observed in Heinz 3402 (12.46 ± 0.69, mg/100 g, fresh weight), followed by Leader (7.87 ± 0.72) and Perfectpeel (2.70 ± 0.68). Antioxidant capacity was significantly correlated with the lycopene and rutin content. Finally, PCA was applied to chemotype data-sets, confirming both mineral content and functional antioxidant compounds as useful markers to unambiguously identify these high-lycopene content varieties.     

The antihypertensive effect of ethyl acetate extract of radish leaves in spontaneously hypertensive rats

Radish (Raphanus sativus L.) is a cruciferous vegetable, and its leaves have antioxidant and anticancer properties. This study was conducted to evaluate the effects of ethyl acetate extracts from radish leaves on hypertension in 11-week-old spontaneously hypertensive rats (SHRs). The SHRs were randomly divided into 3 groups of 6 rats each on the basis of initial systolic blood pressure (SBP) and were treated with oral administration of radish leaf extract (0, 30, or 90 mg/kg body weight [bw], respectively) for 5 weeks. Six Wistar rats were used as normotensive controls. The amount of the radish leaf extract had no effect on body weight. The SBP of the SHRs showed a decreasing trend with the consumption of the radish leaf extract. In the third week, the SBP of the group fed 90 mg extract/kg bw reduced from 214 mmHg to 166 mmHg and was significantly lower than that of the normotensive and hypertensive controls. The extract did not show a significant effect on the angiotensin-converting enzyme (ACE) activity in the serum, kidney, and lung. The extract increased the concentration of NO in serum and the activities of antioxidant enzymes such as glutathione peroxidase and catalase in red blood cells (RBCs). The serum concentrations of Na⁺ and K⁺ were not significantly different between all groups. However, the fecal concentrations of Na⁺ and K⁺ increased; the fecal concentrations of Na⁺ and K⁺ for the normotensive and hypertensive controls were not different. Urinary excretion of Na⁺ was higher in the normotensive Wistar rats than in the SHRs, while that of K⁺ was not significantly different. These findings indicate that consumption of radish leaves might have had antihypertensive effects in SHRs by increasing the serum concentration of NO and fecal concentration of Na⁺ and enhancing antioxidant activities.     

Iron plaque formation on roots of different rice cultivars and the relation with lead uptake

The relationships between lead (Pb) uptake and iron/manganese plaque formation on rice roots were investigated with three cultivars. The results showed that the rice cultivars with indica consanguinity were more sensitive to soil Pb stress than the cultivar with japonica consanguinity. Pb concentrations and distribution ratios in root tissues were in the order: Shanyou 63>Yangdao 6>Wuyunjing 7, but Pb and Fe concentrations and distribution ratios in the plaques showed a reverse order. Mn concentrations and distribution ratios in the plaques of Wuyunjing 7 were significantly higher (P<0.01 or 0.05) than those of Shanyou 63 and Yangdao 6. The results indicate that iron/manganese plaque on rice root can provide a barrier to soil Pb stress. The plaque will increase sequestration of Pb on rice root surface and in the rhizosphere, providing a means of external exclusion of soil Pb to some extent.     

Nutritional composition of Rainbow papaya, the first commercialized transgenic fruit crop

Rainbow papaya (Carica papaya L.) is a genetically engineered (GE) cultivar with resistance to papaya ringspot virus (PRSV). This cultivar currently accounts for about 70% of Hawaii's papaya acreage. The nutritional composition of Rainbow papaya and a non-transgenic control were analyzed to address GE food safety concerns regarding the potential for altered nutritional composition and altered expression of inherent allergens and toxic proteins. Rainbow papaya fruit were analyzed at three ripening stages and the data compared to that of a non-transgenic papaya which shares a similar pedigree. No differences were observed between GE and non-GE papaya for 36 nutrients at any of the tested fruit ripeness stages. However, vitamin A was higher and calcium levels were lower in the GE fruit. The GE fruit showed higher levels of protein and papain at the earliest stage of ripening (color break), but in ripened fruit these differences were insignificant. Benzyl isothiocyanate (BITC) levels were very low and similar for both Rainbow and the non-transgenic control fruit at all ripeness stages. Our data show that the contents of nutrients, BITC and papain of GE Rainbow papaya are within the range of those of non-GE papaya and that the Rainbow cultivar is substantially similar to the non-GE cultivar.     

Minerals and trace elements in a collection of wheat landraces from the Canary Islands

Phosphorous, Na, K, Ca, Mg, Fe, Cu, Zn, Mn and Se were determined in eighteen wheat landraces from Canary Islands and a commercial cultivar. The mean values obtained were: P (2370 ± 333 mg/kg); Na (102 ± 52 mg/kg); K (4363 ± 386 mg/kg); Ca (351 ± 62 mg/kg); Mg (1163 ± 155 mg/kg); Fe (40.0 ± 5.5 mg/kg); Cu (2.68 ± 0.93 mg/kg); Zn (32.1 ± 2.9 mg/kg); Mn (22.1 ± 3.5 mg/kg); Se (67.7 ± 40.4 μg/kg). There were differences (P < 0.05) in the mineral and trace element between the analyzed cultivars, and between the species and subspecies of wheats. Therefore, the Triticum aestivum species had higher mean P, Mg, Fe, Cu, Zn and Mn concentrations, and a lower mean Se concentration, than Triticum turgidum species. The environment and agronomic practices could affect the genetic information of the seeds determining changes in the mineral and trace element composition. The wheat represents an excellent source of Mn (over 100% of recommended dietary intake, RDI), as well as P, Mg, Fe, Cu, Zn and Se (20–50% of RDI). Many correlations were found between the parameters analyzed emphasizing the Cu–Mg (r = 0.691) and Cu–Mn (r = 0.619) correlations. Linear discriminant analysis allowed a good differentiation of the wheat samples according to the cultivar (89.7% of wheat samples classified correctly).     

Characterization of chia seed oils obtained by pressing and solvent extraction

The oil yield, fatty acid composition and the physicochemical and quality characteristics of chia crude seed oils obtained by pressing and solvent extraction were determined. The extraction methods assayed influenced significantly the oil yield, obtaining about 30% more oil by solvent than by pressing. The main fatty acids ranked in the following order of abundance: α-linolenic acid (α Ln) > linoleic acid (L) > oleic acid (O) ≈ palmitic acid (P) > stearic acid (S) for both extraction systems. The n-3/n-6 FA ratio of chia oils ranged from 3.18 to 4.18, being markedly higher than that reported for other vegetable oils. The main triacylglycerols were: αLnαLnαLn > αLnαLnL > αLnLL > αLnαLnP > αLnLO ∼ αLnLP, which represent about 87-95% of the total content of these compounds. The quality and composition of some minor constituents of chia seed oils were influenced by the extraction process. Oils presented a moderate content of bioactive components, such as tocopherols, polyphenols, carotenoids and phospholipids; the high unsaturation level determined their low oxidative stability.     

Oxalate content of purslane leaves and the effect of combining them with yoghurt or coconut products

The total oxalate of purslane (Portulaca oleracea L.) leaves grown in a green house was 1072.7 ± 23.2 mg/100 g dry matter (DM) while the level was 1234.1 ± 37.5 for leaves grown in shaded light in the same greenhouse. Lightly cooking the leaves grown in full and shaded light had no effect on soluble or insoluble content of the leaf tissue. Overall, leaves grown in shaded light contained higher levels of insoluble oxalates leading to an increased amount of total calcium within the leaves being bound to oxalate compared to the levels in the leaves grown in full light in the greenhouse. Addition of yoghurt, coconut milk or coconut cream to the raw leaves had the effect of reducing the overall oxalate content of the mixture by simple dilution. However, the addition of yoghurt to raw purslane leaves significantly reduced the soluble oxalate content of the mixture. The soluble oxalate content of the raw leaves was 53.0% which reduced to 10.7% when yoghurt was added to the leaves. Addition of coconut milk or coconut cream to fresh purslane leaves, while they both gave the mixture an acceptable taste, had no effect on reducing the percentage soluble oxalate content of the mixture.     

Pinot Noir grape colour related phenolics as affected by leaf removal treatments in the Vipava Valley

Wine colour depends directly on the quantitative and qualitative profiles of anthocyanins, flavonols and hydroxycinnamic acids, as well as on their involvement in polymerization and co-pigmentation reactions. Pinot Noir is a grape variety with a low natural colour potential, often combined with problems of low colour stability for its wines during aging. The influence of leaf removal timing on grape colour related polyphenols was observed during maturation and at harvest time for the 2009 vintage “real scenario” conditions in two different vineyards in the Vipava Valley (Slovenia), known for its warm but windy climate. With different timing of leaf removal, a different microclimate can be achieved, with different light exposure for the specific plant tissue and different temperatures on berry surfaces. Phenolic compounds were characterised and quantified using HPLC/Vis. Three classes of polyphenols were affected by leaf removal timing, but with differing intensities and with some differences between locations. Hydroxycinnamic acids were affected only slightly, mostly at the beginning of the maturation period. Anthocyanins were significantly affected, while flavonols were the most affected by treatments in both vineyards and their concentration was clearly related to modification of the light within the canopy.     

Analysis of flavor volatiles of glutinous rice during cooking by combined gas chromatography–mass spectrometry with modified headspace solid-phase microextraction method

A combined gas chromatography–mass spectrometry (GC–MS) with modified headspace solid-phase microextraction (SPME) method, with the headspace inside a designed sampling apparatus instead of in a vial, has been employed for the analysis of the flavor volatiles of glutinous rice during cooking. All of the free flavor volatiles, the bound flavor components and the compounds formed by the thermal decomposition of the non-volatile constituents existing in rice could be liberated during the cooking process. Therefore, a broad range of the flavor volatiles of glutinous rice during cooking could be extracted, concentrated and identified in a single headspace SPME/GC–MS run. Altogether, 96 volatile compounds were identified, of which 27 components have not been previously reported in rice. A total of 86, 90 and 94 peaks, respectively, were assigned to Tatsukomochi, Kinunohada and Miyakoganemochi during cooking. The volatile components identified in the three glutinous rice cultivars during cooking belong to the chemical classes of aldehydes, ketones, alcohols and heterocyclic compounds, as well as fatty acids and esters, phenolic compounds, hydrocarbons, etc.     

Attenuating effect of Lactobacillus brevis G101 on the MSG symptom complex in a double-blind,placebo-controlled study

BACKGROUND/OBJECTIVESLactobacillus brevis G101 suppresses the absorption of monosodium glutamate (MSG) from the intestine into the blood in mice. Therefore, the attenuating effect of orally administered G101 on monosodium glutamate (MSG) symptom complex was investigated in humans.MATERIALS/METHODSCapsules (300 mg) containing Lactobacillus brevis G101 (1×10¹⁰ CFU/individual) or maltodextrin (placebo) was orally administered in 30 respondents with self-recognized monosodium glutamate (MSG) symptom complex for 5 days and the rice with black soybean sauce containing 6 g MSG (RBSM) was ingested 30 min after the final administration. Thereafter, the MSG symptom complex (rated on a 5-point scale: 1, none; 5, strong) was investigated in a double blind placebo controlled study. The intensity of the MSG symptom complex was significantly reduced in respondents of the G101 intake group (2.87 ± 0.73) compared to that in those treated with the placebo (3.63 ± 1.03) (P = 0.0016). Respondents in the placebo group exhibited more of the various major conditions of the MSG symptom complex than in the G101 intake group. Although there was no significant difference in the appearance time of the MSG symptom complex between subjects orally administered G101 and those administered the placebo, its disappearance in < 3 h was observed in 69.9% of subjects in the G101 treatment group and in 38.0% of subjects in the placebo group (P = 0.0841).CONCLUSIONSOral administration of Lactobacillus brevis G101 may be able to reduce the intensity of the MSG symptom complex.     

Protective effect of a DNA vaccine containing an open reading frame with homology to an ABC-type transporter present in the genomic island 3 of Brucella abortus in BALB/c mice

The immunogenicity of a DNA vaccine containing an open reading frame (ORF) of genomic island 3 (GI-3), specific for Brucella abortus and Brucella melitensis, has been examined. Intramuscular injection of plasmid DNA carrying the open reading frame with homology to an ABC-type transporter (pV278a) into BALB/c mice elicited both humoral and cellular immune responses. Mice injected with pV278a had a dominant immunoglobulin G2a (IgG2a) response. This DNA vaccine elicited a T-cell-proliferative response and induced significant levels of interferon gamma (INF-γ) upon restimulation with recombinant 278a protein. Upon stimulation with an appropriate recombinant protein or crude Brucella protein, the vaccine did not induce IL-4, suggesting a typical T-helper (TH1) response. Furthermore, the vaccine induced protection in BALB/c mice when challenged with the virulent strain Brucella abortus 2308. Taken together, these data suggest that DNA vaccination offers an improved delivery of the homologous of an ABC-type transporter antigen, and provides the first evidence of a protective effect of this antigen in the construction of vaccines against B. abortus.     

The effect on the bones of condensed phosphate when used as food additives: Its Importance in Relation to Preventive Medicine

Based on the fact that chemical products such as binding agents are produced by mixing three kinds of phosphates with different ratios, we mixed metaphosphate, polyphosphate and pyrophosphate. Each was made to Na-phosphate, K-phosphate, and Ca-phosphate and each was mixed with commercial feeds so that the content of P would be approximately 0.1, 0.15, 0.3, 0.4, 0.6 and 1.0%. The prepared pellets were given to ICR, CF # 1 and AKR strains of mice at 29 days of age for 680 days and observations were made through this experimental period at different stages. The observations were also carried out on the mice administered with the experimental feeds for 1.5 months from 9 to 10.5 months of age. The observations were compared with those of the control group at all times. As a result, plasma 1 α, 25 (OH)₂ D₃ and P levels were always significantly higher in the phosphate administered groups relative to the control. Urine P and Fe increased while urine Ca decreased in the phosphate-treated groups. The effect of phosphates on the bones was studied taking soft X-ray pictures of hind legs and applying microdensitometry to them. Through these observations we recognized thinning of the cortex of bones, reduction of marrow trabecules and development of osteophyte. Histological observations disclosed that changes in knee joint tissues were apparent; that is, a decrease in or an irregular loss of the number of cells in superficial, intermediate, and radial strata of the joint cartilage, proliferation of subchondral bone, and the development of osteophytes were noted. As for muscles, diameters of musclar fibers became smaller; in particular, type II fibers showed greater shrinkage. Regarding kidneys, swelling and atrophy of glomerular capillaries, proliferation of mesangial cells, nephroselerosis, swelling, thinning, and loss of tubular epithelium, interstitial tissue inflammation, development of cylindruria, and deposition of calcium were observed. All these changes seem to be a particularly advanced aspect of the changes which are more pronounced with increasing dose and age. These changes were found even in the group administered with the feed containing 0.1% phosphorus, and, these changes were dependent on the concentration level of P. It was observed that administration to older subjects for a short term (1.5 months) produced effects stronger than those to younger subjects administered for a long term (10.5 months). The effects of condensed Ca-phosphate on bones were similar to those of condensed Na- and K-phosphates, and, hence, it was supposed that these effects were caused by phosphate radicals. An erratum to this article is available at .     

Randomized Controlled Trial Examining the Effects of Fish Oil and Multivitamin Supplementation on the Incorporation of n-3 and n-6 Fatty Acids into Red Blood Cells

The present randomized, placebo-controlled, double-blind, parallel-groups clinical trial examined the effects of fish oil and multivitamin supplementation on the incorporation of n-3 and n-6 fatty acids into red blood cells. Healthy adult humans (n = 160) were randomized to receive 6 g of fish oil, 6 g of fish oil plus a multivitamin, 3 g of fish oil plus a multivitamin or a placebo daily for 16 weeks. Treatment with 6 g of fish oil, with or without a daily multivitamin, led to higher eicosapentaenoic acid (EPA) composition at endpoint. Docosahexaenoic acid (DHA) composition was unchanged following treatment. The long chain LC n-3 PUFA index was only higher, compared to placebo, in the group receiving the combination of 6 g of fish oil and the multivitamin. Analysis by gender revealed that all treatments increased EPA incorporation in females while, in males, EPA was only significantly increased by the 6 g fish oil multivitamin combination. There was considerable individual variability in the red blood cell incorporation of EPA and DHA at endpoint. Gender contributed to a large proportion of this variability with females generally showing higher LC n-3 PUFA composition at endpoint. In conclusion, the incorporation of LC n-3 PUFA into red blood cells was influenced by dosage, the concurrent intake of vitamin/minerals and gender.     

Reducing effect of the Chinese medicinal herb, Salvia miltiorrhiza, on alcohol self-administration in Sardinian alcohol-preferring rats

The dried roots of Salvia miltiorrhiza are highly valued in Chinese folk medicine for use in the prevention and treatment of a series of ailments. Previous studies have demonstrated that administration of standardized extracts of S. miltiorrhiza selectively reduced excessive alcohol drinking and relapse-like drinking in selectively bred Sardinian alcohol-preferring (sP) rats. The present study was designed to extend these findings on the “anti-alcohol” properties of S. miltiorrhiza extracts to operant procedures of oral alcohol self-administration. Two independent groups of sP rats were trained to lever-respond on an FR4 schedule of reinforcement for alcohol (15%, v/v) or sucrose (1–3%, w/v) in daily 30 min sessions. Once responding had stabilized, rats were tested under the fixed ratio 4 (FR4) schedule of reinforcement (index of alcohol reinforcing properties) and the progressive ratio (PR) schedule of reinforcement (index of alcohol motivational properties). Treatment with S. miltiorrhiza extract (0, 50, 100, and 200 mg/kg, intragastrically [i.g.]) markedly reduced lever responding for alcohol, amount of self-administered alcohol, and breakpoint for alcohol (defined as the lowest response requirement not achieved in the PR experiment). No dose of S. miltiorrhiza extract altered any parameter of sucrose self-administration. These results a) demonstrate that treatment with S. miltiorrhiza extract selectively reduced the reinforcing and motivational properties of alcohol in sP rats and b) extend to operant procedures of alcohol self-administration previous data on the “anti-alcohol” effects of S. miltiorrhiza extracts. These data strengthen the notion that novel pharmacological approaches for treatment of alcohol use disorders may stem from natural substances.     

Proximate, unsaponifiable lipid and fatty acid composition of bogue (Boops boops) and horse mackerel (Trachurus trachurus) from the Italian trawl fishery

This study was aimed at evaluating the nutritional properties of bogue (Boops boops) and horse mackerel (Trachurus trachurus), fish species present throughout the Mediterranean and highly represented in the catch of the Italian trawl fishery. In Italy such fish species are scarcely known by consumers and are not sought-after on the market, especially when the fish are small. Proximates, unsaponifiables and fatty acid profile of bogue and horse mackerel caught by trawlers in different seasons of the year along the southern Adriatic coast of Italy were evaluated. Results showed that both species were characterised by good protein contents (18–20 g/100 g) and low lipid (1–2 g/100 g) and cholesterol (50–70 mg/100 g) levels at any season considered. Along with α-tocopherol (0.45–0.70 mg/100 g), δ-tocopherol was found at a lower level (0.04–0.09 mg/100 g) while no detectable amounts of the γ-isomer were observed in either species. The fatty acid profiles of horse mackerel and bogue exhibited a seasonal fluctuation. They were characterised by high proportions of polyunsaturated fatty acids (PUFA), ranging from about 30% of total fatty acids in summer to more than 40% of total fatty acids in spring and winter, and by high n-3/n-6 PUFA ratio values (4.4–6.8 for bogue, 7.4–8.2 for horse mackerel). On a quantitative basis, in bogue total n-3 PUFA amounted to 0.29–0.43 g out of 0.33–0.54 g of total PUFA per 100 g wet fillet. In horse mackerel total n-3 PUFA ranged between 0.37 g and 0.43 g/100 g and total PUFA between 0.41 and 0.49 g/100 g wet fillet. The atherogenic (0.46–0.63) and thrombogenic (0.22–0.38) indexes calculated on bogue and horse mackerel were comparable to those of fish species of higher commercial value. The nutritional properties of these underutilised species may be considered comparable to those of other low-fat fish species from the wild and also from aquaculture present on the Italian market and meeting consumers’ preferences.     

Differential effect of CD4Foxp3 T-regulatory cells on the B and T helper cell responses to influenza virus vaccination

The T-regulatory (T-reg) cells restrict the T-cell functions in various viral infections including influenza infection. However little is known about the effect of T-regs in influenza vaccination. Herein, we found that immunization of BALB/c mice with a prototype of UV-inactivated influenza PR8/A/34 virus vaccine expanded the CD4⁺Foxp3⁺ T-reg pool and fostered the development of virus-specific CD4⁺Foxp3⁺ T-reg cells. Increasing the size of Foxp3⁺ T-reg pool did not alter the primary PR8-specific B-cell response, but it did suppress the primary and memory PR8-specific T helper responses induced by vaccination. In contrast, the vaccination-induced T helper cell response was augmented in the absence of CD4⁺Foxp3⁺ T-reg cells. Since CD4 T helper cells contribute to anti-influenza protection, therapeutic “quenching” of T-reg function prior to vaccination may enhance the efficacy of influenza vaccination.     

Effects of varying antigens and adjuvant systems on the immunogenicity and safety of investigational tetravalent human oncogenic papillomavirus vaccines: Results from two randomized trials

Background

A prophylactic human papillomavirus (HPV) vaccine targeting oncogenic HPV types in addition to HPV-16 and -18 may broaden protection against cervical cancer. Two Phase I/II, randomized, controlled studies were conducted to compare the immunogenicity and safety of investigational tetravalent HPV L1 virus-like particle (VLP) vaccines, containing VLPs from two additional oncogenic genotypes, with the licensed HPV-16/18 AS04-adjuvanted vaccine (control) in healthy 18–25 year-old women.

Methods

In one trial (NCT00231413), subjects received control or one of 6 tetravalent HPV-16/18/31/45 AS04 vaccine formulations at months (M) 0,1,6. In a second trial (NCT00478621), subjects received control or one of 5 tetravalent HPV-16/18/33/58 vaccines formulated with different adjuvant systems (AS04, AS01 or AS02), administered on different schedules (M0,1,6 or M0,3 or M0,6).

Results

One month after the third injection (Month 7), there was a consistent trend for lower anti-HPV-16 and -18 geometric mean antibody titers (GMTs) for tetravalent AS04-adjuvanted vaccines compared with control. GMTs were statistically significantly lower for an HPV-16/18/31/45 AS04 vaccine containing 20/20/10/10 μg VLPs for both anti-HPV-16 and anti-HPV-18 antibodies, and for an HPV-16/18/33/58 AS04 vaccine containing 20/20/20/20 μg VLPs for anti-HPV-16 antibodies. There was also a trend for lower HPV-16 and -18-specific memory B-cell responses for tetravalent AS04 vaccines versus control. No such trends were observed for CD4⁺ T-cell responses. Immune interference could not always be overcome by increasing the dose of HPV-16/18 L1 VLPs or by using a different adjuvant system. All formulations had acceptable reactogenicity and safety profiles. Reactogenicity in the 7-day post-vaccination period tended to increase with the introduction of additional VLPs, especially for formulations containing AS01.

Conclusions

HPV-16 and -18 antibody responses were lower when additional HPV L1 VLPs were added to the HPV-16/18 AS04-adjuvanted vaccine. Immune interference is a complex phenomenon that cannot always be overcome by changing the antigen dose or adjuvant system.