Development and Validation of a Reversed-Phase Liquid Chromatography Method for the Simultaneous Determination of Indole-3-Acetic Acid, Indole-3-Pyruvic Acid, and Abscisic Acid in Barley (Hordeum vulgare L.)

A simple, sensitive, precise, and specific reverse HPLC method was developed and validated for the determination of plant hormones in barley (Hordeum vulgare L.). The method includes extraction in aqueous organic solvent followed by solid-phase extraction, sample evaporation, and reversed-phase HPLC analysis in a general purpose UV-visible (abscisic acid (ABA)) and fluorescence detection (indole-3-acetic acid (IAA) and indole-3-pyruvic acid (IPA)), high-performance liquid chromatography system. The separation was carried out on Zorbax Eclipse XDB C8 column (150  ×  4.6  mm I.D) with a mobile phase composed of methanol and 1% acetic acid (60 : 40 v/v) in isocratic mode at a flow rate of 1 ml min−1. The detection was monitored at 270 nm (ABA) and at 282 nm (Ex) and 360 nm (Em) (IAA, IPA). The developed method was validated in terms of accuracy, precision, linearity, limit of detection, limit of quantification, and robustness. The determined validation parameters are in the commonly acceptable ranges for that kind of analysis

Broad Prebiotic Potential of Non-starch Polysaccharides from Oats (Avena sativa L.): an in vitro Study

Prebiotics inducing the growth or activity of beneficial intestinal bacteria – probiotics producing short-chain fatty acids (SCFA) have lately received wide recognition for their beneficial influence on host intestinal microbiota and metabolic health. Some non-starch polysaccharides (NSP) are defined as prebiotics and oats being one of richest sources of NSP in grains are considered as potentially having prebiotic effect. However, information on fermentation of specific NSP of oats is limited. Moreover, bacterial cross-feeding interactions in which fermentation of prebiotics is involved is poorly characterized. Here, we report the exploration of new candidates for the syntrophic bacterial interactions and fermentability of oat non-starch polysaccharides (NSP). The results obtained by differentiating composition, viscosity and concentration of oats NSP in fermentation medium showed that Bacillus licheniformis pre-digests oat NSP, degrades high viscosity of oat β-glucan and makes hemicellulose easier to access for other bacteria. Because of fermentation, B. licheniformis produces lactic and succinic acids, which further can be used by other bacteria for cross-feeding and SCFA production

CHARACTERIZATION OF ANTIOXIDANT ACTIVITY AND TOTAL PHENOLIC COMPOUND CONTENT OF BIRCH OUTER BARK EXTRACTS USING MICRO PLATE ASSAY

In modern plants, 2.7 to 2.8 m3 of solid volume veneer blocks are consumed to produce 1 m3 of plywood. After the hydrothermal treatment and debarking of blocks, waste bark is obtained, which makes up 12.5% of the wood mass, while 16-20% of bark is composed of birch outer bark (BOB). Recalculating, BOB makes up 2.0-3.4% of the veneer log mass. Bark is currently burned in boiler houses that is not rational. BOB contains large amount of valuable extractives (up to 34% from o.d. BOB) consisting of various secondary metabolites such as terpenes, flavonoids, hydrocarbons, polyphenols, tannins etc. BOB extractives exhibit antioxidant properties as well as wound-healing and anti-inflammatory activity. The objective of this paper was to compare amount of total phenolic content (TPC) and antiradical activity (ARA) in ethanol extracts from silver birch (Betula pendula Roth.) outer bark using micro plate assay. Among 11 fractions, fraction of average BOB had the highest DPPH free radical scavenging activity with IC50 of 39.28 μg/mL and the highest TPC 7.42 ± 0.52 g GAE/100 g of dry extract

Differential disease resistance response in the barley necrotic mutant nec1

<p>Abstract</p> <p>Background</p> <p>Although ion fluxes are considered to be an integral part of signal transduction during responses to pathogens, only a few ion channels are known to participate in the plant response to infection. CNGC4 is a disease resistance-related cyclic nucleotide-gated ion channel. <it>Arabidopsis thaliana </it>CNGC4 mutants <it>hlm1 </it>and <it>dnd2 </it>display an impaired hypersensitive response (HR), retarded growth, a constitutively active salicylic acid (SA)-mediated pathogenesis-related response and elevated resistance against bacterial pathogens. Barley CNGC4 shares 67% aa identity with AtCNGC4. The barley mutant <it>nec1 </it>comprising of a frame-shift mutation of CNGC4 displays a necrotic phenotype and constitutively over-expresses <it>PR-1</it>, yet it is not known what effect the <it>nec1 </it>mutation has on barley resistance against different types of pathogens.</p> <p>Results</p> <p><it>nec1 </it>mutant accumulated high amount of SA and hydrogen peroxide compared to parental cv. Parkland. Experiments investigating <it>nec1 </it>disease resistance demonstrated positive effect of <it>nec1 </it>mutation on non-host resistance against <it>Pseudomonas syringae </it>pv. <it>tomato </it>(<it>Pst</it>) at high inoculum density, whereas at normal <it>Pst </it>inoculum concentration <it>nec1 </it>resistance did not differ from wt. In contrast to augmented <it>P. syringae </it>resistance, penetration resistance against biotrophic fungus <it>Blumeria graminis </it>f. sp. <it>hordei </it>(<it>Bgh</it>), the causal agent of powdery mildew, was not altered in <it>nec1</it>. The <it>nec1 </it>mutant significantly over-expressed race non-specific <it>Bgh </it>resistance-related genes <it>BI-1 </it>and <it>MLO</it>. Induction of <it>BI-1 </it>and <it>MLO </it>suggested putative involvement of <it>nec1 </it>in race non-specific <it>Bgh </it>resistance, therefore the effect of <it>nec1</it>on <it>mlo-5</it>-mediated <it>Bgh </it>resistance was assessed. The <it>nec1</it>/<it>mlo-5 </it>double mutant was as resistant to <it>Bgh </it>as <it>Nec1</it>/<it>mlo-5 </it>plants, suggesting that <it>nec1 </it>did not impair <it>mlo-5 </it>race non-specific <it>Bgh </it>resistance.</p> <p>Conclusions</p> <p>Together, the results suggest that <it>nec1 </it>mutation alters activation of systemic acquired resistance-related physiological markers and non-host resistance in barley, while not changing rapid localized response during compatible interaction with host pathogen. Increased resistance of <it>nec1 </it>against non-host pathogen <it>Pst </it>suggests that <it>nec1 </it>mutation may affect certain aspects of barley disease resistance, while it remains to be determined, if the effect on disease resistance is a direct response to changes in SA signaling.</p

Primary culture of avian embryonic heart forming region cells to study the regulation of vertebrate early heart morphogenesis by vitamin A

Copyright: Copyright 2014 Elsevier B.V., All rights reserved.Background: Important knowledge about the role of vitamin A in vertebrate heart development has been obtained using the vitamin A-deficient avian in ovo model which enables the in vivo examination of very early stages of vertebrate heart morphogenesis. These studies have revealed the critical role of the vitamin A-active form, retinoic acid (RA) in the regulation of several developmental genes, including the important growth regulatory factor, transforming growth factor-beta2 (TGFβ2), involved in early events of heart morphogenesis. However, this in ovo model is not readily available for elucidating details of molecular mechanisms determining RA activity, thus limiting further examination of RA-regulated early heart morphogenesis. In order to obtain insights into RA-regulated gene expression during these early events, a reliable in vitro model is needed. Here we describe a cell culture that closely reproduces the in ovo observed regulatory effects of RA on TGFβ2 and on several developmental genes linked to TGFβ signaling during heart morphogenesis. Results: We have developed an avian heart forming region (HFR) cell based in vitro model that displays the characteristics associated with vertebrate early heart morphogenesis, i.e. the expression of Nkx2.5 and GATA4, the cardiogenesis genes, of vascular endothelial growth factor (VEGF-A), the vasculogenesis gene and of fibronectin (FN1), an essential component in building the heart, and the expression of the multifunctional genes TGFβ2 and neogenin (NEO). Importantly, we established that the HFR cell culture is a valid model to study RA-regulated molecular events during heart morphogenesis and that the expression of TGFβ2 as well as the expression of several TGFβ2-linked developmental genes is regulated by RA. Conclusions: Our findings reported here offer a biologically relevant experimental in vitro system for the elucidation of RA-regulated expression of TGFβ2 and other genes involved in vertebrate early cardiovascular morphogenesis.publishersversionPeer reviewe

DOSAGE OF CISPLATIN AND RELATED TOXICITY OF CANCER PATIENTS BASED ON BODY FAT AND MUSCLE MASS

In oncology practice, the human body surface area (BSA) is used for the calculation of the dose of chemotherapy. The human BSA is determined by derived formulas, but it is not directly linked to the pharmacokinetics of the drugs. Obesity alters the disposition of drugs pharmacokinetics, which should be considered when prescribing medications in this patient population. Failure to adjust doses in obesity may result in increased toxicity. The aim of this study was to assess predictors of an increase in the level of trygliceride considered to be among the cardiovascular risk factors. Method. The study involved 118 oncology patients with various tumor localizations. The patients underwent anthropometric measurements (height, weight, circumference, fat fold thickness, the distance between the epicondyls above the joint), and the BSA (according to the Mosteller formula). Patients who received cisplatin chemotherapy were analyzed in serum triglyceride levels before and after the treatment. Results. A comparison of female and male subsamples revealed no significant in variance between sexes and no significant differences in initial triglyceride, triglyceride change, and in the age between groups. Body fat mass was relatively higher in females, while body muscle mass and bone mass were relatively higher in males. There was a difference in the cisplatin dose, with a higher dose for males than for females. The results demonstrate that the higher cisplatin dose and higher body fat mass are significant predictors of an increase in triglyceride for both sexes

DOSAGE OF CISPLATIN AND RELATED TOXICITY OF CANCER PATIENTS BASED ON BODY FAT AND MUSCLE MASS

In oncology practice, the human body surface area (BSA) is used for the calculation of the dose of chemotherapy. The human BSA is determined by derived formulas, but it is not directly linked to the pharmacokinetics of the drugs. Obesity alters the disposition of drugs pharmacokinetics, which should be considered when prescribing medications in this patient population. Failure to adjust doses in obesity may result in increased toxicity. The aim of this study was to assess predictors of an increase in the level of trygliceride considered to be among the cardiovascular risk factors. Method. The study involved 118 oncology patients with various tumor localizations. The patients underwent anthropometric measurements (height, weight, circumference, fat fold thickness, the distance between the epicondyls above the joint), and the BSA (according to the Mosteller formula). Patients who received cisplatin chemotherapy were analyzed in serum triglyceride levels before and after the treatment. Results. A comparison of female and male subsamples revealed no significant in variance between sexes and no significant differences in initial triglyceride, triglyceride change, and in the age between groups. Body fat mass was relatively higher in females, while body muscle mass and bone mass were relatively higher in males. There was a difference in the cisplatin dose, with a higher dose for males than for females. The results demonstrate that the higher cisplatin dose and higher body fat mass are significant predictors of an increase in triglyceride for both sexes

The Cultivation of Chelidonium majus L. Increased the Total Alkaloid Content and Cytotoxic Activity Compared with Those of Wild-Grown Plants

Funding Information: Funding: This research was funded by European Regional Development Fund project “Innovative solutions for growing technologies and applications of spring medicinal and aromatic plants” (Grant No. 1.1.1.1/18/A/ 043). Publisher Copyright: © 2021 by the authors. Licensee MDPI, Basel, Switzerland.The effect of cultivation practises on both the phytochemical profile and biological activity of aqueous ethanol extracts of Chelidonium majus L. was studied. Extracts were prepared from aerial parts of the same plant population collected in the wild and grown under organic farming conditions. Both qualitative and quantitative analyses of alkaloids and flavonoid derivatives were performed by LC/MS methods, and the cytotoxicity of lyophilised extracts was studied in B16-F10, HepG2, and CaCo-2 cells. Coptisine was the dominant alkaloid of extracts prepared from wild-grown plants, whereas after cultivation, chelidonine was the most abundant alkaloid. The total alkaloid content was significantly increased by cultivation. Ten flavonol glycoconjugates were identified in C. majus extracts, and quantitative analysis did not reveal significant differences between extracts prepared from wild-grown and cultivated specimens. Treatment with C. majus extracts resulted in a dose-dependent increase in cytotoxicity in all three cell lines. The extracts prepared from cultivated specimens showed higher cytotoxicity than the extracts prepared from wild-grown plants. The strongest cytotoxic effect of cultivated C. majus was observed in B16-F10 cells (IC50 = 174.98 ± 1.12 µg/mL). Cultivation-induced differences in the phytochemical composition of C. majus extracts resulted in significant increases in the cytotoxic activities of the preparations.publishersversionPeer reviewe

Exploring the Potential of Supercritical Fluid Extraction of Matricaria chamomilla White Ray Florets as a Source of Bioactive (Cosmetic) Ingredients

Funding Information: This research was funded by the European Regional Development Fund project “Biorefinery approach for the development of bioactive cosmetic ingredients from byproducts of medicinal plant processing and plant cell cultivation” (No. 1.1.1.1/19/A/075). Publisher Copyright: © 2023 by the authors.Aromatic and medicinal plants are a great source of useful bioactive compounds for use in cosmetics, drugs, and dietary supplements. This study investigated the potential of using supercritical fluid extracts obtained from Matricaria chamomilla white ray florets, a kind of industrial herbal byproduct, as a source of bioactive cosmetic ingredients. Response surface methodology to optimize the supercritical fluid extraction process by analyzing the impact of pressure and temperature on yield and the main bioactive compound groups were used. High-throughput 96-well plate spectrophotometric methods were used to analyze the extracts for total phenols, flavonoids, tannins, and sugars, as well as their antioxidant capacity. Gas chromatography and liquid chromatography-mass spectrometry was used to determine the phytochemical composition of the extracts. The extracts were also analyzed for antimicrobial activity, cytotoxicity, phototoxicity, and melanin content. Statistical analysis was performed to establish correlations between the extracts and develop models to predict the targeted phytochemical recovery and chemical and biological activities. The results show that the extracts contained a diverse range of phytochemical classes and had cytotoxic, proliferation-reducing, and antimicrobial activities, making them potentially useful in cosmetic formulations. This study provides valuable insights for further research on the uses and mechanisms of action of these extracts.publishersversionPeer reviewe

Wild-Grown and Cultivated Glechoma hederacea L. : Chemical Composition and Potential for Cultivation in Organic Farming Conditions

Funding Information: Funding: This work was supported by the European Regional Development Fund project “Innovative solutions for growing technologies and applications of spring medicinal and aromatic plants” (Nr. 1.1.1.1/18/A/043). Publisher Copyright: © 2022 by the authors. Licensee MDPI, Basel, Switzerland.Glechoma hederacea L. is a medicinal plant that is known in traditional medicine for its anti-inflammatory, antibacterial, antiviral, and anticancer properties. This study evaluated the potential for commercial production of G. hederacea and compared the chemical composition and activity of 70% ethanol extracts and steam-distilled essential oils from wild-grown and cultivated G. hederacea collected in different harvesting periods. The main compounds identified in the 70% ethanol extracts were phenolic acids (chlorogenic and rosmarinic acids) and flavonoid O-glycosides. The essential oil varied in the three accessions in the range of 0.32-2.98 mL/kg -1 of dry weight. The extracts possessed potent antioxidant and anti-inflammatory properties in LPS-treated bone-marrow-derived macrophages. The results of flow cytometry show that extracts from different vegetation periods reduced the conversion of macrophages to the proinflammatory phenotype M1. The chemical composition varied the most with the different harvesting periods, and the most suitable periods were the flowering and vegetative phases for the polyphenolic compounds and essential oils, respectively. G. hederacea can be successfully grown under organic farming conditions, and cultivation does not significantly affect the chemical composition and biological activity compared to wild-grown plants.publishersversionPeer reviewe