In Vitro and In Vivo Anti-Inflammatory Effects of Cannabidiol Isolated from Novel Hemp (Cannabis sativa L.) Cultivar Pink Pepper.

Cannabis sativa L. contains more than 80 cannabinoids, among which cannabidiol (CBD) is the main neuroactive component. We aimed to investigate the anti-inflammatory efficacy of CBD in vitro and in vivo isolated from "Pink pepper", a novel hemp cultivar, by repeating the method of selecting and cultivating individuals with the highest CBD content. We investigated the effects of CBD on inflammatory markers elevated by lipopolysaccharide (LPS) treatment in RAW 264.7 mouse macrophage cells through Western blot and RT-PCR. In addition, we confirmed these effects through the ELISA of inflamed paw tissue of a λ-carrageenan-induced mouse edema model that received an oral administration of CBD. CBD inhibited the LPS-induced phosphorylation of NF-κB and MAPK in RAW 264.7 and exhibited anti-inflammatory effects by participating in these pathways. In our in vivo study, we confirmed that CBD also inhibited the inflammatory mediators of proteins extracted from edematous mouse paw tissue. These results show that CBD isolated from "Pink pepper" exhibits potent anti-inflammatory effects. These anti-inflammatory effects of CBD have pharmacological and physiological significance, highlighting the industrial value of this novel cultivar.

An Enhanced Water Solubility and Stability of Anthocyanins in Mulberry Processed with Hot Melt Extrusion.

Mulberry fruits are rich sources of anthocyanins that exhibit beneficial biological activity. These anthocyanins become instable in an aqueous media, leading to their low bioavailability. In this study, a colloidal dispersion was produced by processing mulberry samples with hot-melt extrusion. In this process, hydrophilic polymer matrices were used to disperse the compound in an aqueous media. Mulberry samples were processed with hot-melt extrusion and in the presence of an ionization agent and sodium alginate to form mulberry-extrudate solid formulations. The particle size of mulberry-extrudate solid formulations decreased, while the total phenol content, the total anthocyanin content, and solubility increased. Fourier transform infrared spectroscopy (FT-IR) revealed that mulberry-extrudate solid formulations now contained new functional groups, such as -COOH group. We investigated whether mulberry-extrudate solid formulations had a positive impact on the stability of anthocyanins. The non-extrudate mulberry sample and mulberry-extrudate solid formulations were incubated with a simulated gastric fluid system and an intestinal fluid system. The number of released anthocyanins was determined with HPLC. We found that anthocyanins were released rapidly from non-extrudate mulberry extract. Mulberry-extrudate solid formulations contained a large number of available anthocyanins even after being incubated for 180 min in the intestinal fluid system. Thus, hot-melt extrusion enhanced water solubility and stability of anthocyanins with the prolonged release.

Conversion Characteristics of Some Major Cannabinoids from Hemp (Cannabis sativa L.) Raw Materials by New Rapid Simultaneous Analysis Method.

This study was carried out to develop a high-performance liquid chromatography method for short-time analysis of the main cannabinoids in the inflorescence of hemp (Cannabis sativa L.). We also performed decarboxylation of the raw material using our advanced analysis technique. In this study, the UV spectrum was considered to analyze each of the four common cannabinoids, solvents, and samples, where the uniform elution of acidic cannabinoids without peak tailing and acids was tested. Optimal results were obtained when readings were taken at a wavelength of 220 nm using water and methanol containing trifluoroacetic acid as mobile phases in a solvent gradient system. The established conditions were further validated by system suitability, linearity, precision, detection limit, and quantitation limit tests. The decarboxylation index (DT50) confirmed that Δ9-THCA decarboxylated faster than CBDA, and both maintained a linear relationship with time and temperature. In addition, the loss of cannabidiol was better prevented during the decarboxylation process in the natural state than in the extracted state.

Characterization of New Polyphenolic Glycosidic Constituents and Evaluation of Cytotoxicity on a Macrophage Cell Line and Allelopathic Activities of Oryza sativa.

Four new constituents, as 5, 7-dihydroxy-4'-methoxyflavonol-3-O-ß-d-arabinopyranosyl-(2''→1''')-O-ß-d-arabinopyrnosyl-2'''-O-3'''', 7''''-dimethylnonan-1''''-oate (1), 5-hydroxy-7, 4'-dimethoxyflavone-5-O-α-d-arabinopyranosyl-(2"→1''')-O-α-d-arabinopyranosyl-2'''-O-3'''', 7''''-dimethylnonan-1''''-oate (2), 5-hydroxy-7, 4'-dimethoxyflavone-5-O-ß-d-arabinofuranosyl-(2"→1''')-O-ß-d-arabinopyranosyl-2'''-O-lanost-5-ene (3) and 4',4''-diferuloxy feruloyl-O-α-d-arabinopyranosyl-(2a→1b)-O-α-d-arabinopyranosyl-(2b→1c)-O-α-d-arabinopyranosyl-(2c→1d)-O-α-d-arabinopyranosyl-(2d→1e)-O-α-d-arabinopyranosyl-2e-3''', 7'''-dimethylnonan-1'''-oate (4), along with three known compounds (5⁻7) were isolated from Oryza sativa leaves and straw. The structures of new and known compounds were elucidated by 1D (¹H and 13C NMR) and 2D NMR spectral methods, viz: COSY, HMBC, and HSQC aided by mass techniques and IR spectroscopy. The cytotoxicity of these constituents was assessed by using (RAW 264.7) mouse macrophage cell line, and allelopathic effects of compounds (1⁻7) on the germination and seedling growth characteristics such as seedling length and root length of barnyardgrass (Echinochloa oryzicola) were evaluated. Significant inhibitory activity was exhibited by compounds comprising flavone derivatives such as (1⁻3) on all of seed germination characteristics. The allelopathic effect of flavone derivatives were more pronounced on seedling length and root length than the germination characteristics. The higher concentration of flavone derivatives showed stronger inhibitory effects, whereas the lower concentrations showed stimulatory effects in some cases.

Fucoidan from Fucus vesiculosus protects against alcohol-induced liver damage by modulating inflammatory mediators in mice and HepG2 cells.

Fucoidan is an l-fucose-enriched sulfated polysaccharide isolated from brown algae and marine invertebrates. In this study, we investigated the protective effect of fucoidan from Fucus vesiculosus on alcohol-induced murine liver damage. Liver injury was induced by oral administration of 25% alcohol with or without fucoidan (30 mg/kg or 60 mg/kg) for seven days. Alcohol administration increased serum aspartate aminotransferase and alanine aminotransferase levels, but these increases were suppressed by the treatment of fucoidan. Transforming growth factor beta 1 (TGF-ß1), a liver fibrosis-inducing factor, was highly expressed in the alcohol-fed group and human hepatoma HepG2 cell; however, the increase in TGF-ß1 expression was reduced following fucoidan administration. Treatment with fucoidan was also found to significantly reduce the production of inflammation-promoting cyclooygenase-2 and nitric oxide, while markedly increasing the expression of the hepatoprotective enzyme, hemeoxygenase-1, on murine liver and HepG2 cells. Taken together, the antifibrotic and anti-inflammatory effects of fucoidan on alcohol-induced liver damage may provide valuable insights into developing new therapeutics or interventions.

Effect of Light Quality on Seed Potato (Solanum tuberose L.) Tuberization When Aeroponically Grown in a Controlled Greenhouse.

A plant factory equipped with artificial lights is a comparatively new concept when growing seed potatoes (Solanum tuberosum L.) for minituber production. The shortage of disease-free potato seed tubers is a key challenge to producing quality potatoes. Quality seed tuber production all year round in a controlled environment under an artificial light condition was the main purpose of this study. The present study was conducted in a plant factory to investigate the effects of distinct spectrum compositions of LEDs on potato tuberization when grown in an aeroponic system. The study was equipped with eight LED light combinations: L1 = red: blue: green (70 + 25 + 5), L2 = red: blue: green (70 + 20 + 10), L3 = red: blue: green (70 + 15 + 15), L4 = red: blue: green (70 + 10 + 20), L5 = red: blue: far-red (70 + 25 + 5), L6 = red: blue: far-red (70 + 20 + 10), L7 = red: blue: far-red (70 + 15 + 15), L8 = red: blue: far-red (70 + 10 + 20), and L9 = natural light with 300 µmol m-2 s-1 of irradiance, 16/8 h day/night, 65% relative humidity, while natural light was used as the control treatment. According to the findings, treatment L4 recorded a higher tuber number (31/plant), tuber size (>3 g); (9.26 ± 3.01), and GA3 content, along with better plant growth characteristics. Moreover, treatment L4 recorded a significantly increased trend in the stem diameter (11.08 ± 0.25), leaf number (25.32 ± 1.2), leaf width (19 ± 0.81), root length (49 ± 2.1), and stolon length (49.62 ± 2.05) compared to the control (L9). However, the L9 treatment showed the best performance in plant fresh weight (67.16 ± 4.06 g) and plant dry weight (4.46 ± 0.08 g). In addition, photosynthetic pigments (Chl a) (0.096 ± 0.00 mg g-1, 0.093 ± 0.00 mg g-1) were found to be the highest in the L1 and L2 treatments, respectively. However, Chl b and TCL recorded the best results in treatment L4. Finally, with consideration of the plant growth and tuber yield performance, treatment L4 was found to have the best spectral composition to grow quality seed potato tubers.

Antisense-overexpression of the MsCOMT gene induces changes in lignin and total phenol contents in transgenic tobacco plants.

Initially, we isolated the caffeic acid O-methyltransferase (COMT) gene from Miscanthus sinensis (accession number HM062766.1). Next, we produced transgenic tobacco plants with down-regulated COMT gene expression to study its control of total phenol and lignin content and to perform morphological analysis. These transgenic plants were found to have reduced PAL and ascorbate peroxidases expression, which are related to the phenylpropanoid pathway and antioxidant activity. The MsCOMT-down-regulated plants had decreased total lignin in the leaves and stem compared with control plants. Reduced flavonol concentrations were confirmed in MsCOMT-down-regulated transgenic plants. We also observed a morphological difference, with reduced plant cell number in transgenic plants harboring antisense MsCOMT. The transgenic tobacco plants with down-regulated COMT gene expression demonstrate that COMT plays a crucial role related to controlling lignin and phenol content in plants. Also, COMT activity may be related to flavonoid production in the plant lignin pathway.

Anti-Obesity Effects of the Larval Powder of Steamed and Lyophilized Mature Silkworms in a Newly Designed Adult Mouse Model.

Recently, "mature" silkworms (MS) of Bombix mori have been considered a potential nutraceutical, with a number of health benefits reported for steamed and lyophilized MS powder (SMSP). However, no obesity-related effects have been reported for SMSP. In the present study, anti-obesity effects of SMSP were investigated in adult mice in vivo, aged 12 weeks at the onset of SMSP treatment, fed a normal diet (ND) and a high-fat diet (HFD), respectively, for 12 weeks. SMSP significantly suppressed body weight gain, intra-abdominal adipose tissue, and food efficiency in both ND-fed and HFD-fed adult mice. In addition, SMSP significantly decreased food intake and liver weight in HFD-fed mice, indicating that SMSP suppressed appetite and simultaneously reduced the conversion of feed into body weight in HFD-fed mice. SMSP also significantly lowered the serum levels of glucose, triglyceride, total cholesterol, low-density lipoprotein cholesterol, asparagine transaminase, alanine transaminase, and alkaline phosphatase. However, SMSP had no significant effect on the weights of the kidney, spleen, or thymus or the serum levels of blood urea nitrogen and creatinine. Taken together, the above results suggest that SMSP has potent anti-obesity effects and is safe for long-term use as a potential therapeutic and/or nutraceutical in both obese patients and non-obese individuals.

Cellular Morphology and Transcriptome Comparative Analysis of Astragalus membranaceus Bunge Sprouts Cultured In Vitro under Different LED Light.

Astragalus membranaceus, the major components of which are saponins, flavonoids, and polysaccharides, has been established to have excellent pharmacological activity. After ginseng, it is the second most used medicinal plant. To examine the utility of A. membranaceus as a sprout crop for plant factory cultivation, we sought to establish a functional substance control model by comparing the transcriptomes of sprouts grown in sterile, in vitro culture using LED light sources. Having sown the seeds of A. membranaceus, these were exposed to white LED light (continuous spectrum), red LED light (632 nm, 1.58 µmol/m2/s), or blue LED light (465 nm, 1.44 µmol/m2/s) and grown for 6 weeks; after which, the samples were collected for transcriptome analysis. Scanning electron microscopy analysis of cell morphology in plants exposed to the three light sources revealed that leaf cell size was largest in those plants exposed to red light, where the thickest stem was observed in plants exposed to white light. The total number of genes in A. membranaceus spouts determined via de novo assembly was 45,667. Analysis of differentially expressed genes revealed that for the comparisons of blue LED vs. red LED, blue LED vs. white LED, and red LED vs. white LED, the numbers of upregulated genes were 132, 148, and 144, respectively. Binding, DNA integration, transport, phosphorylation, DNA biosynthetic process, membrane, and plant-type secondary cell wall biogenesis were the most enriched in the comparative analysis of blue LED vs. red LED, whereas Binding, RNA-templated DNA biosynthetic process, DNA metabolic process, and DNA integration were the most enriched in the comparative analysis of blue vs. white LED, and DNA integration and resolution of meiotic recombination intermediates were the most enrichment in the comparison between red LED vs. white LED. The GO term associated with flavonoid biosynthesis, implying the functionality of A. membranaceus, was the flavonoid biosynthetic process, which was enriched in the white LED vs. red LED comparison. The findings of this study thus indicate that different LED light sources can differentially influence the transcriptome expression pattern of A. membranaceus sprouts, which can provide a basis for establishing a flavonoid biosynthesis regulation model and thus, the cultivation of high-functional Astragalus sprouts.

Prokinetic Activity of Mulberry Fruit, Morus alba L.

The fruit of Morus alba L. (MAF) has been consumed as a food worldwide. MAF has also been widely used in traditional medicine for thousands of years in East Asia, and its diverse bioactivities have been reported in numerous publications. However, no prokinetic activity has been reported for MAF or its components. In the present study, therefore, we investigated the effects of MAF on gastrointestinal motor function by measuring the intestinal transit rate (ITR) of Evans blue in mice in vivo. The ITR values accelerated by MAF were significantly higher than those accelerated by cisapride or metoclopramide, suggesting that MAF has potential as a new prokinetic agent to replace cisapride and metoclopramide. We also investigated the effects of MAF on myogenic and neurogenic contractions in human intestinal smooth muscles by measuring spontaneous contractions of smooth muscle strips, smooth muscle contractions induced by neural stimulation, and migrating motor complexes from intestinal segments in the human ileum and sigmoid colon in situ. MAF increased both myogenic and neurogenic contractions to enhance ileal and colonic motility in the human intestine. Taken together, these results indicate that MAF enhanced intestinal motility by increasing both myogenic and neurogenic contractions, thereby accelerating the ITR.

In Vitro and In Vivo Anti-Inflammatory Potential of Cannabichromene Isolated from Hemp.

Cannabichromene (CBC), a non-psychoactive cannabinoid found in Cannabis sativa, has recently been shown to possess several medicinal properties. However, how CBC produces anti-inflammatory effects and the mechanisms of this remain poorly studied. Therefore, we extracted and purified the CBC from the Cannabis sativa cv. pink pepper (hemp cultivar). The efficacy of CBC in reducing inflammation in RAW 264.7 macrophages and a λ-carrageenan-induced mouse model was then evaluated. CBC had no cytotoxicity up to a concentration of 20 µM and inhibited nitric oxide production by approximately 50% at a concentration of 20 µM. In addition, CBC treatment significantly inhibited causes of inflammation such as inducible nitric oxide synthase (iNOS), interleukin-1ß (IL-1ß), IL-6, and tumor necrosis factor-α (TNF-α) at both the mRNA and protein levels. Moreover, CBC suppressed LPS-stimulated inflammation in RAW 264.7 cells by downregulating the nuclear factor kappa B (NF-kB) and mitogen-activated protein kinase pathways (MAPK). Furthermore, our in vivo experiments confirmed that the λ-carrageenan-induced increase in the levels of the cytokines iNOS, IL-1ß, and IL-6 was abrogated following treatment with CBC. Therefore, CBC has potential anti-inflammatory effects and may be useful for preventing or treating inflammation.

Hot-Melt Extrusion Enhances Antioxidant Effects of Mulberry on Probiotics and Pathogenic Microorganisms.

Mulberry is a rich source of anthocyanins (ACNs) known to possess biological activities. However, these ACNs are unstable in high pH, heat, and aqueous environments with a low bioavailability. In this study, a colloidal dispersion was prepared by hot melt extrusion with proper excipients. In this process, a hydrophilic polymer matrix was used to confirm the stability of the compound in high pH, high temperature, and aqueous media. It was confirmed that the particle size and the polydispersity index value were reduced, thereby improving the solubility. In vitro release studies revealed that the extrudate had a sustained release compared to a non-extruded product. As a result of measuring changes of intestinal microorganisms (Lacticaseibacillus rhamnosus, Pediococcus pentosaceus, Escherichia coli, Enterococcus faecalis, and Staphylococcus aureus), contents of probiotics were found to be increased whereas contents of pathogenic microorganisms were decreased. Thus, hot-melt extrusion could enhance the stability of ACN with prolonged release. The processed formulation exhibited probiotic properties and antimicrobial activities against pathogenic intestinal microflora.

Cannabinoid accumulation in hemp depends on ROS generation and interlinked with morpho-physiological acclimation and plasticity under indoor LED environment.

Manipulation of growth and development of cannabis (Cannabis sativa L.) has received considerable interest by the scientific community due to its high value in medicinal and recreational use worldwide. This study was conducted to investigate the effects of LED spectral changes on reactive oxygen species (ROS) and cannabinoid accumulation by provoking growth, pigmentation, photosynthesis, and secondary metabolites production of cannabis grown in an indoor environment. After three weeks of vegetative growth under greenhouse condition, plants were further grown for 90 days in a plant factory treated with 4 LED light compositions with a canopy-level photosynthetic photon flux density (PPFD) of 300 µmol m-2 s-1 for 16 h. Photosynthetic pigments and photosynthetic rate were linearly increased up to 60 days and then sharply decreased which was found most prominent in L3: MB 240 (Red 85% + Blue 15%) and L4: PF 240 (Red 70% + Blue 30%) LED light compositions. A high concentration of H2O2 was also observed in L3 and L4 treatments which provoked lipid peroxidation in later growth stage. In addition, higher accumulation of cannabinoid was observed under L4 treatment in most cases. It is also evident that higher ROS created a cellular stress in plant as indicated by higher osmolyte synthesis and enzyme activity which initiate quick maturation along with higher cannabinoids accumulation in cannabis plant. Therefore, it can be concluded that ROS metabolism has a crucial role in morpho-physiological acclimation and cannabinoid accumulation in hemp plants. The findings of this study provide further insight on the use of LED light to maximize the production of cannabinoid.

Immunotoxicity activity from various organic solvents extract of Allium genus from South Korea against Aedes aegypti L.

Species of the Allium genus, used in functional foods for the treatment of improving gastritis and heart failures, were collected and petroleum ether, chloroform, and ethyl acetate extracts were obtained by sonication. The Allium thumbergii chloroform extracts have a significant toxic effect against early fourth-stage larvae of Aedes aegypti L. with an LC(50) value of 2.4 µg/mL and an LC(90) value of 6.3 µg/mL. The Allium victorialis var. platyphyllum petroleum ether extracts have a significant toxic effect against early fourth-stage larvae of A. aegypti L. with an LC(50) value of 1.2 µg/mL and an LC(90) value of 2.1 µg/mL. Also, the Allium sacculiferum chloroform extracts have a significant toxic effect against early fourth-stage larvae of A. aegypti L. with an LC(50) value of 1.8 µg/mL and an LC(90) value of 4.7 µg/mL. The above data indicate that major compounds may play a more important role in the toxicity of A. victorialis var. platyphyllum petroleum ether extracts.

Exogenous Putrescine Enhances Salt Tolerance and Ginsenosides Content in Korean Ginseng (Panax ginseng Meyer) Sprouts.

The effect of exogenously applied putrescine (Put) on salt stress tolerance was investigated in Panax ginseng. Thirty-day-old ginseng sprouts were grown in salinized nutrient solution (150 mM NaCl) for five days, while the control sprouts were grown in nutrients solution. Putrescine (0.3, 0.6, and 0.9 mM) was sprayed on the plants once at the onset of salinity treatment, whereas control plants were sprayed with water only. Ginseng seedlings tested under salinity exhibited reduced plant growth and biomass production, which was directly interlinked with reduced chlorophyll and chlorophyll fluorescence due to higher reactive oxygen species (hydrogen peroxide; H2O2) and lipid peroxidation (malondialdehyde; MDA) production. Application of Put enhanced accumulation of proline, total soluble carbohydrate, total soluble sugar and total soluble protein. At the same time, activities of antioxidant enzymes like superoxide dismutase, catalase, ascorbate peroxidase, guaiacol peroxidase in leaves, stems, and roots of ginseng seedlings were increased. Such modulation of physio-biochemical processes reduced the level of H2O2 and MDA, which indicates a successful adaptation of ginseng seedlings to salinity stress. Moreover, protopanaxadiol (PPD) ginsenosides enhanced by both salinity stress and exogenous Put treatment. On the other hand, protopanaxatriol (PPT) ginsenosides enhanced in roots and reduced in leaves and stems under salinity stress condition. In contrast, they enhanced by exogenous Put application in all parts of the plants for most cases, also evidenced by principal component analysis. Collectively, our findings provide an important prospect for the use of Put in modulating salinity tolerance and ginsenosides content in ginseng sprouts.

Cannabinoids Accumulation in Hemp (Cannabis sativa L.) Plants under LED Light Spectra and Their Discrete Role as a Stress Marker.

Hemp adaptability through physiological and biochemical changes was studied under 10 LED light spectra and natural light in a controlled aeroponic system. Light treatments were imposed on 25 days aged seedlings for 16 h daily (300 µmol m-2 s-1) for 20 days. Plant accumulated highest Cannabidiol (CBD) in R7:B2:G1 light treatment, with relatively higher photosynthetic rate and lower reactive oxygen species, total phenol content, total flavonoid content, DPPH radical scavenging capacity, and antioxidant enzymatic activities. Tetrahydrocannabinol (THC) also accumulated at a higher level in white, R8:B2, and R7:B2:G1 light with less evidence of stress-modulated substances. These results indicated that CBD and THC have no or little relation with light-mediated abiotic stress in hemp plants. On the contrary, Tetrahydrocannabinolic acid (THCA) was accumulated higher in R6:B2:G1:FR1 and R5:B2:W2:FR1 light treatment along with lower photosynthetic rate and higher reactive oxygen species, total phenol content, total flavonoid content, DPPH radical scavenging capacity, and antioxidant enzymatic activities. However, Cannabidiolic acid (CBDA) was accumulated higher in R6:B2:G1:FR1 light treatment with higher stress-modulated substances and lower physiological traits. CBDA was also accumulated higher in R8:B2 and R7:B2:G1 light treatments with less evidence of stress-modulated substances. Besides, Greenlight influenced CBD and CBDA synthesis where FR and UV-A (along with green) play a positive and negative role in this process. Overall, the results indicated that the treatment R7:B2:G1 enhanced the medicinal cannabinoids most, and the role of THCA as a stress marker is more decisive in the hemp plant than in other cannabinoids under attributed light-mediated stress.

Production of lactosucrose from sucrose and lactose by a levansucrase from Zymomonas mobilis.

Lactosucrose (4(G)-beta-D-galactosylsucrose) is an oligosaccharide consisting of galactose, glucose, and fructose. In this study, we prepared lactosucrose from lactose and sucrose using a levansucrase derived from Zymomonas mobilis. Optimum conditions for lactosucrose formation were 23 degrees C, pH 7.0, 18.0% (w/v) lactose monohydrate, and 18% (w/v) sucrose as substrates, and 1 unit of enzyme/ml of reaction mixture. Under these conditions, the lactosucrose conversion efficiency was 28.5%. The product was purified and confirmed to be O-beta-D-galactopyranosyl-(1-->4)-O-beta-D-glucopyranosyl- (1-->2)-beta-D-fructofuranoside, or lactosucrose. A mixed-enzyme system containing a levansucrase and a glucose oxidase was applied in order to increase the efficiency of lactose and sucrose conversion to lactosucrose, which rose to 43.2% as s result.

Bio- Fortification of Angelica gigas Nakai Nano-Powder Using Bio-Polymer by Hot Melt Extrusion to Enhance the Bioaccessibility and Functionality of Nutraceutical Compounds.

Angelica gigas Nakai (AGN) is a popular traditional herbal medicine which has been used to alleviate various human diseases in Korea since ancient times. However, the low bioaccessibility of the nutraceutical compounds of AGN results in a poor water solubility, thereby limiting bioavailability. In this regard, a ternary AGN-biopolymer-plasticizer composite (AGNC) was developed to enhance the bioaccessibility of nutraceutical compounds from extrudate AGN formulations manufactured by hot melt extrusion (HME). The AGNC was prepared with extrudate AGN (EAGN) using different hydroxypropyl methylcellulose (HPMC) biopolymers (5% w/w) viz.: hypromellose phthalate (HP), hypromellose (AN), and hypromellose (CN) along with acetic acid (AA) (0.1 M, 20% w/v) as a plasticizer. The non-extrudate fresh AGN (FAGN) powder was used as a control. The physicochemical properties of the extrudate formulations and control were characterized by differential scanning calorimetry (DSC) and Fourier-transform infrared spectroscopy (FTIR). DSC analysis showed a lower enthalpy (ΔH) (12.22 J/g) and lower glass transition temperature (Tg) (41 °C) in HP-AA-EAGN compared to the control. FTIR confirmed the physical crosslinking between AGN and biopolymer in the extrudate composite and demonstrated that some functional groups formed viz., -OH and -CH2. The obtained result also shows that the particle size was reduced by 341 nm, and solubility was increased by 65.5% in HP-AA-EAGN compared to the control (1499 nm, 29.4%, respectively). The bioaccessibility of the total phenolic content and the total flavonoids-including decursin (D) and decursinol angelate (DA)-were significantly higher in HP-AA-EAGN compared to the control. The 2,2-diphenyl-1 picryl hydrazyl (DPPH) free radical scavenging capacity and ferric reducing antioxidant power assay (FRAP) indicated that the HP-AA-EAGN formulation preserves a greater antioxidant profile than the other formulations. Finally, it is summarized that the addition of acidified HP biopolymer increased the bioaccessibility, functionality, and improved the physicochemical properties of nutraceutical compounds in the extrudate AGN formulation.

Effect of Different Processing Methods on the Accumulation of the Phenolic Compounds and Antioxidant Profile of Broomcorn Millet (Panicum miliaceum L.) Flour.

Broomcorn millet (Panicum miliaceum L.) is an important nutritious ancient minor-cereal food crop. However, this crop is little explored in the food processing arena to improve its functionality. In this context, different processing methods were applied to enhance the secondary compounds of broomcorn millet. Four different individual methods such as roasting, steaming, puffing, and extrusion were applied at 110 °C to enhance the functional attributes of millet flour. It was observed that the significantly highest content of total phenolic (TP) (670 mg/100 g of ferulic acid equivalent) and total flavonoid (TF) (391 mg/100 g of rutin equivalent ) was attained in the roasted whole millet followed by steaming (315 mg/100 g, 282 mg/100 g), puffing (645 mg/100 g, 304 mg/100 g), extrusion (455 mg/100 g, 219 mg/100 g), and control (295 mg/100 g, 183 mg/100 g). The chromatographic analysis showed a greater content of single phenolic acids such as syringic acid, gallic acid, 4-hydroxy benzoic acid, ferulic acid, sinapic acid, and catechin in roasted millet compared to control, and the content of each acid was higher in whole millet than dehulled. Results also indicated that the content of ferulic acid was relatively higher among the quantified single phenolic acid from broomcorn millet. Likewise, in comparison with dehulled millet, the roasted whole millet showed higher total antioxidant capacity, measured by the 2,2-diphenyl-1 picryl hydrazyl (DPPH), the ferric reducing antioxidant power assay (FRAP), the phosphomolybdenum method (PPMD), and the hydroxyl radical scavenging capacity (HRSC) method. Lastly, it is concluded that the roasting method should be taken into consideration in the processing of broomcorn millet to enhance the content of nutraceutical compounds and improve its functionality.

Expression of a functional type-I chalcone isomerase gene is localized to the infected cells of root nodules of Elaeagnus umbellata.

A putative type-I chalcone isomerase (CHI) cDNA clone EuNOD-CHI was previously isolated from the root nodule of Elaeagnus umbellata [Kim et al. (2003)]. To see if it encodes a functional CHI, we ectopically overexpressed it in the Arabidopsis (Arabidopsis thaliana) transparent testa 5 (tt5) mutant, which is defective in naringenin production and has yellow seeds due to proanthocyanidin deficiency. Ectopic overexpression of EuNOD-CHI resulted in recovery of normal seed coat color. Naringenin produced by CHI from naringenin chalcone was detected in the transgenic lines like in the wild-type, whereas it was absent from the tt5 mutant. We conclude that EuNOD-CHI encodes a functional type-I CHI. In situ hybridization revealed that EuNOD-CHI expression is localized to the infected cells of the fixation zone in root nodules.