Comparison of Yield Characteristics, Chemical Composition, Lignans Content and Antioxidant Potential of Experimentally Grown Six Linseed (Linum usitatissimum L.) Cultivars.

Linseed represents a rich source of nutritional, functional and health-beneficial compounds. Nevertheless, the chemical composition and content of bioactive compounds may be quite variable and potentially affected by various factors, including genotype and the environment. In this study, the proximate chemical composition, lignans content and antioxidant potential of six experimentally grown linseed cultivars were assessed and compared. A diagonal cultivation trial in the University of South Bohemia Experimental Station in Ceské Budejovice, Czech Republic, was established in three subsequent growing seasons (2018, 2019 and 2020). The results showed that the cultivar and growing conditions influenced most studied parameters. The lack of precipitation in May and June 2019 negatively affected the seed yield and the level of secoisolariciresinol diglucoside but did not decrease the crude protein content, which was negatively related to the oil content. The newly developed method for lignans analysis allowed the identification and quantification of secoisolariciresinol diglucoside and matairesinol. Their content correlated positively with the total polyphenol content and antioxidant assays (DPPH and ABTS radical scavenging activity), indicating the significant contribution to the biofunctional properties of linseed. On the other hand, we did not detect minor linseed lignans, pinoresinol and lariciresinol. The results of this study showed the importance of cultivar and growing conditions factors on the linseed chemical composition and the lignans content, determining its nutritional and medicinal properties.

Formation of Previously Undescribed Δ7-Phytosterol Oxidation Products and Tocopherylquinone Adducts in Pumpkin Seed Oil during Roasting, Screw-Pressing, and Simulated Culinary Processing at Elevated Temperatures.

The influence of pumpkin seed roasting conditions (110-140 °C) and screw-pressing on the formation of previously undescribed Δ7-phytosterol oxidation products and tocopherylquinone adducts with nucleophilic phosphatidylethanolamine species was investigated. The roasting process of pumpkin seed paste at a temperature above 120 °C for 30 min considerably enhanced the formation of Δ7-oxysterols. Targeted analysis [electron impact mass spectrometry (MS), 1D-nuclear magnetic resonance] led to the identification of five novel markers of pumpkin paste roasting, among which (3ß,5α,22E,24S)-stigmasta-7,22-dien-6-one-3-ol (6-oxo-α-spinasterol), stereoisomers of (3ß,5α,22E)-7,8-epoxystigmast-22-en-3-ol (7,8-epoxy-α-spinasterol), and (3ß,5α)-22,23-epoxystigmast-7-en-3-ol (7,8-epoxy-α-spinasterol) were reported in edible oils for the first time. Simulated culinary processing provided novel stereoisomers of (3ß,5α,22E)-stigmasta-7,22-dien-3,6-diol, unusual (3ß,5α,22E)-stigmasta-7,22-dien-6,15-dione-3-ol, and (5α,22E)-stigmasta-7,22-dien-3-one accompanied by minor stereoisomers of (3ß,5α)-7,8;22,23-diepoxystigmastan-3-ol. Moreover, a clear relationship between the pumpkin seed oil stability index and synergistic effect of glycerophospholipids with present tocochromanols was found. High-resolution atmospheric pressure chemical ionization-MS experiments clearly demonstrated the formation of various γ-tocopherylquinone adducts with primary amines, namely, octylamine. The mitigation strategy of potentially detrimental oxysterols from pumpkin seed oil included optimization of processing parameters while maintaining the formation of desirable sensory-active compounds.

The Use of a Thermal Process to Produce Black Garlic: Differences in the Physicochemical and Sensory Characteristics Using Seven Varieties of Fresh Garlic.

Black garlic (BG) is a product originating from fresh garlic (FG) and substantially differs in many aspects from FG due to the process called ageing. During this thermal process, the health-promoting properties of FG are enhanced, and the sensory traits are altered. However, very little is known about how the physicochemical properties of different FG varieties affect these properties of BG. Thus, the aim of this study was to investigate the influence of seven FG varieties subjected to the thermal process on the physicochemical parameters of BG. To prepare the BG samples, a fifteen-day ageing process involving a temperature gradient ranging from 30 to 82 °C was used. It was found that the antioxidant activity, the total polyphenol content, and the total soluble solids increased during ageing, while the pH level, moisture content, and lightness decreased in all the garlic varieties. The varieties of garlic differed in the studied traits significantly, both before (FG) and after ageing (BG). In the sensory analysis, significant differences between the BG varieties were observed only in the pleasantness of texture, while the remaining sensory descriptors (pleasantness of color, odor, taste and intensity of the garlic aroma, and overall acceptability) were not affected by variety. The correlations suggest that most of the FG's studied parameters in this study do not correlate with the properties of BG and cannot be used for the prediction of the quality of BG. Additionally, HPLC-MS/MS analysis revealed substantial changes in the composition of low molecular compounds.

Oilseed Cake Flour Composition, Functional Properties and Antioxidant Potential as Effects of Sieving and Species Differences.

Oilseed cakes are produced as a by-product of oil pressing and are mostly used as feed. Their use for human consumption is due to the functional properties and benefits for human health. Herein, oilseed cake flours of eight species (flax, hemp, milk thistle, poppy, pumpkin, rapeseed, safflower, sunflower) were sieved into fractions above (A250) and below (B250) 250 µm. The chemical composition, SDS-PAGE profiles, colour, functional properties and antioxidant activities of these flours were evaluated. The B250 fractions were evaluated as being protein and ash rich, reaching crude protein and ash content ranging from 31.78% (milk thistle) to 57.47% (pumpkin) and from 5.0% (flax) to 11.19% (poppy), respectively. A high content of carbohydrates was found in the flours of hemp, milk thistle and safflower with a significant increase for the A250 fraction, with a subsequent relation to a high water holding capacity (WHC) for the A250 fraction (flax, poppy, pumpkin and sunflower). The A250 milk thistle flour was found to have the richest in polyphenols content (TPC) (40.89 mg GAE/g), with the highest antioxidant activity using an ABTS•+ assay (101.95 mg AAE/g). The A250 fraction for all the species exhibited lower lightness than the B250 fraction. The obtained results indicate that sieving oilseed flour with the aim to prepare flours with specific functional characteristics and composition is efficient only in combination with a particular species.

Antifungal and antimicrobial proteins and peptides of potato (Solanum tuberosum L.) tubers and their applications.

Potato proteins are well known for their nutritional, emulsifying, foaming, gel forming or antioxidant properties that all make from them valuable protein source for food industry. Antifungal, antimicrobial and also antiviral properties, described for potato proteins in the review, enrich the possibilities of potato protein usage. Potato proteins were divided into patatin, protease inhibitors and fraction of other proteins that also included, besides others, proteins involved in potato defence physiology. All these proteins groups provide proteins and peptides with antifungal and/or antimicrobial actions. Patatins, obtained from cultivars with resistance to Phytophthora infestans, were able to inhibit spore germination of this pathogen. Protease inhibitors represent the structurally heterogeneous group with broad range of antifungal and antimicrobial activities. Potato protease inhibitors I and II reduced the growth of Phytophthora infestans, Rhizoctonia solani and Botrytis cinerea or of the fungi of Fusarium genus. Members of Kunitz family (proteins Potide-G, AFP-J, Potamin-1 or PG-2) were able to inhibit serious pathogens such as Staphylococcus aureus, Listeria monocytogenes, Escherichia coli or Candida albicans. Potato snakins, defensins and pseudothionins are discussed for their ability to inhibit serious potato fungi as well as bacterial pathogens. Potato proteins with the ability to inhibit growth of pathogens were used for developing of pathogen-resistant transgenic plants for crop improvement. Incorporation of potato antifungal and antimicrobial proteins in feed and food products or food packages for elimination of hygienically risk pathogens brings new possibility of potato protein usage.

Development of Composite Poly(Lactide-co-Glycolide)- Nanodiamond Scaffolds for Bone Cell Growth.

There are relatively few nanotechnologies that can produce nanocomposite scaffolds for cell growth. Electrospinning has emerged as the foremost method of producing nanofibrous biomimetic scaffolds for tissue engineering applications. In this study diamond nanoparticles were integrated into a polymer solution to develop a nanocomposite scaffold containing poly(lactide-co-glycolide) (PLGA) loaded with diamond nanoparticles. To investigate the effect of adding diamond nanoparticles to PLGA scaffolds, primary human mesenchymal stem cells (hMSCs) were seeded on the scaffolds. The cytocompatibility results showed that addition of diamond nanoparticles did not impinge upon cell proliferation, nor was there a cytotoxic cellular response after 9 days in culture. Scanning electron microscopy, transmission electron microscopy, atomic force microscopy and confocal microscopy enabled qualitative characterization of the fibres and revealed cell morphology and number. Furthermore, surface roughness was measured to evaluate diamond nanoparticle modifications, and no significant difference was found between the diamond nanocomposite and pure polymer scaffolds. On the other hand, bright spots on phase images performed by atomic force microscopy suggested a higher hardness at certain points on fibers of the PLGA-nanodiamond composites, which was supported by nanoindentation measurements. This study shows that PLGA nanofibers can be reinforced with nanodiamond without adversely affecting cell behaviour, and thus it sets the foundation for future application of these scaffolds in bone tissue engineering.

Unexpected behavior of some nitric oxide modulators under cadmium excess in plant tissue.

Various nitric oxide modulators (NO donors--SNP, GSNO, DEA NONOate and scavengers--PTIO, cPTIO) were tested to highlight the role of NO under Cd excess in various ontogenetic stages of chamomile (Matricaria chamomilla). Surprisingly, compared to Cd alone, SNP and PTIO elevated Cd uptake (confirmed also by PhenGreen staining) but depleted glutathione (partially ascorbic acid) and phytochelatins PC2 and PC3 in both older plants (cultured hydroponically) and seedlings (cultured in deionised water). Despite these anomalous impacts, fluorescence staining of NO and ROS confirmed predictable assumptions and revealed reciprocal changes (decrease in NO but increase in ROS after PTIO addition and the opposite after SNP application). Subsequent tests using alternative modulators and seedlings confirmed changes to NO and ROS after application of GSNO and DEA NONOate as mentioned above for SNP while cPTIO altered only NO level (depletion). On the contrary to SNP and PTIO, GSNO, DEA NONOate and cPTIO did not elevate Cd content and phytochelatins (PC2, PC3) were rather elevated. These data provide evidence that various NO modulators are useful in terms of NO and ROS manipulation but interactions with intact plants affect metal uptake and must therefore be used with caution. In this view, cPTIO and DEA NONOate revealed the less pronounced side impacts and are recommended as suitable NO scavenger/donor in plant physiological studies under Cd excess.

Variation of antioxidants and secondary metabolites in nitrogen-deficient barley plants.

Barley (Hordeum vulgare cv. Bojos) plants cultured in low nitrogen (N) containing Hoagland solution (20 mg/l) were exposed to N deficiency (-N) over 15 days. Plants revealed relatively high tolerance to total N deficit because shoot length was not altered and dry biomass was depleted by ca. 30% while root length increased by ca. 50% and dry biomass remained unaffected. Soluble proteins and free amino acids decreased more pronouncedly in the roots. Antioxidants (glutathione and ascorbic acid) decreased in the shoots but increased or were not affected in the roots. Ascorbate peroxidase and glutathione reductase activities were depleted in shoots and/or roots while guaiacol peroxidase activity was stimulated in the shoots. In accordance, fluorescence signal of reactive oxygen species (ROS) and nitric oxide was elevated in shoots but no extensive changes were observed in roots if +N and -N treatments are compared. At the level of phenolic metabolites, slight increase in soluble phenols and some phenolic acids and strong elevation of flavonoid homoorientin was found in the shoots but not in the roots. Fluorescence microscopy in terms of detection of phenols is also discussed. We also briefly discussed accuracy of quantification of some parameters owing to discrepancies in the literature. It is concluded that N deficiency induces increase in shoot phenolics but also elevates symptoms of oxidative stress while increase in root antioxidants probably contributes to ROS homeostasis aimed to maintain root development.

Nanofibrous poly(lactide-co-glycolide) membranes loaded with diamond nanoparticles as promising substrates for bone tissue engineering.

BACKGROUND: Nanofibrous scaffolds loaded with bioactive nanoparticles are promising materials for bone tissue engineering. METHODS: In this study, composite nanofibrous membranes containing a copolymer of L-lactide and glycolide (PLGA) and diamond nanoparticles were fabricated by an electrospinning technique. PLGA was dissolved in a mixture of methylene chloride and dimethyl formamide (2:3) at a concentration of 2.3 wt%, and nanodiamond (ND) powder was added at a concentration of 0.7 wt% (about 23 wt% in dry PLGA). RESULTS: In the composite scaffolds, the ND particles were either arranged like beads in the central part of the fibers or formed clusters protruding from the fibers. In the PLGA-ND membranes, the fibers were thicker (diameter 270 ± 9 nm) than in pure PLGA meshes (diameter 218 ± 4 nm), but the areas of pores among these fibers were smaller than in pure PLGA samples (0.46 ± 0.02 µm(2) versus 1.28 ± 0.09 µm(2) in pure PLGA samples). The PLGA-ND membranes showed higher mechanical resistance, as demonstrated by rupture tests of load and deflection of rupture probe at failure. Both types of membranes enabled the attachment, spreading, and subsequent proliferation of human osteoblast-like MG-63 cells to a similar extent, although these values were usually lower than on polystyrene dishes. Nevertheless, the cells on both types of membranes were polygonal or spindle-like in shape, and were distributed homogeneously on the samples. From days 1-7 after seeding, their number rose continuously, and at the end of the experiment, these cells were able to create a confluent layer. At the same time, the cell viability, evaluated by a LIVE/DEAD viability/cytotoxicity kit, ranged from 92% to 97% on both types of membranes. In addition, on PLGA-ND membranes, the cells formed well developed talin-containing focal adhesion plaques. As estimated by the determination of tumor necrosis factor-alpha levels in the culture medium and concentration of intercellular adhesion molecule-1, MG-63 cells, and RAW 264.7 macrophages on these membranes did not show considerable inflammatory activity. CONCLUSION: This study shows that nanofibrous PLGA membranes loaded with diamond nanoparticles have interesting potential for use in bone tissue engineering.