Gallic Acid Research Articles

Whey protein concentrate and skimmed milk powder as encapsulation agents for coffee silverskin extracts processed by spray drying.

We tested the hypothesis that milk proteins, through microencapsulation, guarantee protection against bioactive substances in coffee silverskin extracts. Therefore, the aim of this study was to carry out technological, nutritional and physicochemical characterisation of a coffee silverskin extract microencapsulated using instant skim milk powder and whey protein concentrate as wall materials. The aqueous extract of coffee silverskin was spray-dried using 10% (w/v) skim milk powder and whey protein concentrate. The samples were characterised by determining the water content, water activity, particle size distribution, colour analysis and total phenolic compound content as well as antioxidant activity using 2,2-diphenyl-radical 1-picrylhydrazyl scavenging methods, nitric oxide radical inhibition and morphological analysis. The product showed water activity within a range that ensured greater stability, and the reduced degradation of the dried coffee silverskin extract with whey protein concentrate resulted in better rehydration ability. The luminosity parameter was higher and the browning index was lower for the encapsulated samples than for the pure coffee silverskin extract. The phenolic compound content (29.23 ± 8.39 and 34.00 ± 8.38 mg gallic acid equivalents/g for the coffee silverskin extract using skimmed milk powder and whey protein concentrate, respectively) and the antioxidant activity of the new product confirmed its potential as a natural source of antioxidant phenolic compounds. We conclude that the dairy matrices associated with spray drying preserved the bioactive and antioxidant activities of coffee silverskin extracts.

Nanomedicine: Patuletin-conjugated with zinc oxide exhibit potent effects against Gram-negative and Gram-positive bacterial pathogens.

With the emergence of drug-resistance, there is a need for novel anti-bacterials or to enhance the efficacy of existing drugs. In this study, Patuletin (PA), a flavanoid was loaded onto Gallic acid modified Zinc oxide nanoparticles (PA-GA-ZnO), and evaluated for antibacterial properties against Gram-positive (Bacilluscereus andStreptococcus pneumoniae)and Gram-negative (Samonella entericaand Escherichia coli) bacteria. Characterization of PA, GA-ZnO and PA-GA-ZnO' nanoparticles was accomplished utilizing fourier-transform infrared spectroscopy, efficiency of drug entrapment, polydispersity index, zeta potential, size, and surface morphology analysis through atomic force microscopy. Using bactericidal assays, the results revealed that ZnO conjugation displayed remarkable effects and enhanced Patuletin's effects against both Gram-positive and Gram-negative bacteria, with the minimum inhibitory concentration observed at micromolar concentrations. Cytopathogenicity assays exhibited that the drug-nanoconjugates reduced bacterial-mediated human cell death with minimal side effects to human cells. When tested alone, drug-nanoconjugates tested in this study showed limited toxic effects against human cells in vitro. These are promising findings, but future work is needed to understand the molecular mechanisms of effects of drug-nanoconjugates against bacterial pathogens, in addition to in vivo testing to determine their translational value. This study suggests that Patuletin-loaded nano-formulation (PA-GA-ZnO) may be implicated in a multi-target mechanism that affects both Gram-positive and Gram-negative pathogen cell structures, however this needs to be ascertained in future work.

Nutritional, Phytochemical and Antioxidant Characterizations of Wild Food and Medicinal Fruits Indigenous to Sudan: Sarcocephalus latifolius and Vitex doniana

ABSTRACT Sudan is a rich country in wild plants that play significant roles in securing food and medicine in rural areas, especially. This study explored Sarcocephalus latifolius (African peach) and Vitex doniana (black plum) for their fruits’ proximate composition, minerals, total soluble phenols, phenolic acids, total carotenoids, lutein, β–carotene, α–tocopherol, δ–tocopherol and total antioxidant capacity. African peach fruit and black plum pulp showed total soluble phenols amounting to 16.89 mg gallic acid equivalents (GAE)/g of Dry Weight (DW) and 7.56 mg GAE/g DW, respectively. The paramount phenolic acids were cinnamic acid (895 mg/kg DW) in African peach fruit and protocatechuic acid (1200 mg/kg DW) in black plum pulp. Total antioxidant capacities reached 268 µmoles trolox equivalents (TE)/g DW in African peach and 214 µmoles TE/g DW in black plum, when measured from hydrophilic extracts (HPE) using 2, 2-DiPhenyl-1-PicrylHydrazyl (DPPH) assay. The contents of total carotenoid were 47 mg/kg DW and 22.6 mg/kg DW, lutein reached 18.50 mg/kg DW and 15 mg/kg DW whereas β–carotene showed 0.31 mg/kg DW and 0.23 mg/kg DW in African peach and black plum, respectively. African peach showed α–tocopherol (167 mg/kg DW) whereas black plum showed δ–tocopherol (398 mg/kg DW). From this study, we concluded that African peach fruit and black plum pulp could be good sources of nutrition and antioxidants and have high potential for human health.

PSV-11 In vitro screening of selected compounds to reduce ruminal methane production

Abstract Methane is a natural product of rumen fermentation and emissions from ruminant livestock represent a major environmental concern. Methane is not only a potent greenhouse gas but also represents a loss of energy efficiency for the host. The objective of this study was to perform in vitro screening of a range of compounds with potential anti-methanogenic activity, including polyphenols, flavonoids, fatty acids, amino acids, and salts. We aimed to select compounds conferring up to a 20% reduction in CH4 for further investigations. The substances tested included carvacrol, coumaric acid, coumarin, flavanone, gallic acid, glycerol monolaurate (GML), glycine, methylene blue, naringin, quercetin, quinic acid, resorcinol, rutin, tannic acid, vanillic acid, and zinc acetate. These substances varied in their mechanisms of action and were selected based on known antimicrobial or anti-methanogenic properties. The experiment was designed as a randomized block experiment, with treatments tested in triplicates across four batches (n = 228 samples) using the same treatments and experimental conditions. Fresh rumen contents were manually collected from two rumen-fistulated lactating Holstein cows, 3 h after their morning feeding. Rumen contents were blended at a 2:1 ratio, filtered through a four-layered cheesecloth, and added to McDougall’s solution at a 2:1 ratio. Final media (9 mL) was added to a 20 mL Bellco flask containing 150 mg of total mixed ration (TMR) ground to 1 mm. Each batch also had three flasks without TMR and no treatment (blank flasks), three flasks containing only TMR without treatment (negative control), and three flasks containing TMR and treated with chloroform (positive control), resulting in a total of 19 treatments. The end-products of in vitro batch fermentations were assessed after 24 h of incubation, and samples from each flask were collected for gas composition analysis using gas chromatography. Volatile fatty acids were determined using high-performance liquid chromatography (HPLC). The data were analyzed using R Studio, and the following model was applied: yij = µ + treatmenti + batchj + ℇij. Methane production (measured in parts per million, ppm) increased when GML, naringin, and quercetin were added compared with the negative control. In contrast, the addition of carvacrol, glycine, tannic acid, and methylene blue resulted in a decrease of up to 25% in methane production. These compounds show potential to alter ruminal fermentation and mitigate methane production and will be selected for evaluation of synergistic interactions and effects on the rumen microbial communities.

Comparative investigation on the phenolic compounds and antioxidant capacity of walnut kernel from different drying methods

Drying techniques are being used more and more to extend the shelf life of industrial products. Drying could influnce the content of phenolics in food and their antioxidant activity. This study estimated the effects of different drying methods (freeze drying (FD), gradient hot air drying (GHD), and constant hot air drying (CHD)) on phenolic profiles and antioxidant activities in walnut kernels. With a maximum content of 3.61 mg g−1, GHD was found to be the most effective in preserving total phenols, while CHD and FD had maximum contents of 2.66 mg g−1 and 1.96 mg g−1, respectively. The concentration of most monomeric phenols detected in the kernels increased with temperature, particularly in the free and bound forms. Gallic acid (free form) levels in GHD2 (194.54 µg g−1) were 55.77 and 60.08 times higher, respectively, than in FD and CHD. GHD dried walnuts had higher antioxidant activity than FD and CHD dried walnuts. Furthermore, bioinformatics analysis revealed three key metabolic pathways associated with the mechanisms underlying drying changes. The GHD technique, according to these findings, is a better choice for drying walnut in order to preserve its phenolics and antioxidant activity.Graphical

Analytical method development and dermal absorption of gallic acid, a hair dye ingredient

Analytical method development and dermal absorption of gallic acid, a hair dye ingredient

Evaluation of the effect of organic fertilizers on the morphophysiological parameters and antioxidant compounds of Swiss chard (Beta vulgaris L.).

Objective: To evaluate the effect of organic fertilizers on the morphophysiological parameters and antioxidant compounds of Swiss chard (B. vulgaris L.) var. ‘Rainbow’ grown under greenhouse conditions. Design/Methodology/Approach: A completely randomized design was used with three treatments and 33 repetitions: T1 (poultry manure-based organic fertilizer (T1 AAC)), T2 (coffe pulp-based vermicompost (T2 LC)), and T3 (chemical fertilization (T3 FQ)). Plant height, stem diameter, number of leaves, leaf area, fresh biomass, dry biomass, chlorophyll A, chlorophyll B, total chlorophyll, carotenoids, total polyphenols, DPPH, and yield were evaluated. An analysis of variance and Tukey’s test (α=0.05) were used. Results: For most of the variables evaluated, the best results were obtained with the poultry manure-based organic fertilizer (T1 AAC) treatment. The antioxidant compounds and the quality of Swiss chard were influenced by organic fertilizers, with 0.21 mg Eq. gallic acid g-1 fresh tissue (ft), total chlorophyll of 19.5 µg g-1 (ft), and a yield of 6,150 kg·m2. Study Limitations/Implications: The mineral content of this vegetable was not evaluated. Findings/Conclusions: The use of poultry manure-based organic fertilizers can be an alternative nutritional management for Swiss chard cultivation.

Phytochemical Characterization, Antioxidant, and Antimicrobial Activity of the Vegetative Buds from Romanian Spruce, Picea abies (L.) H. Karst.

This study aims to investigate the vegetative buds from Picea abies (spruce), naturally found in a central region of Romania, through a comprehensive analysis of the chemical composition to identify bioactive compounds responsible for pharmacological properties. Using HPLC/derivatization technique of GC-MS and quantitative spectrophotometric assays, the phenolic profile, and main components of an ethanolic extract from the buds were investigated. The essential oil was characterized by GC-MS. Moreover, the antioxidant activity with the DPPH method, and the antimicrobial activity were tested. Heavy metal detection was performed by graphite furnace atomic absorption spectrometry. The main components of the alcoholic extract were astragalin, quercetin, kaempferol, shikimic acid, and quinic acid. A total content of 25.32 ± 2.65 mg gallic acid equivalent per gram of dry plant (mg GAE/g DW) and of 10.54 ± 0.083 mg rutin equivalents/g of dry plant (mg RE/g DW) were found. The essential oil had D-limonene, α-cadinol, δ-cadinene, 13-epimanool, and δ-3-carene as predominant components. The spruce vegetative buds exhibited significant antioxidant activity (IC50 of 53 μg/mL) and antimicrobial effects against Staphylococcus aureus. Furthermore, concentrations of heavy metals Pb and Cd were below detection limits, suggesting that the material was free from potentially harmful contaminants. The results confirmed the potential of this indigenous species to be used as a source of compounds with pharmacological utilities.

Characterization and molecular docking of sustainable wine lees and gelatin-based emulsions: innovative fat substitution.

The present study aimed to determine how various amounts (0.00, 0.58, 1.52 and 4.50 g 100 g-1) of wine lees (WL), which contains numerous essential components, impact the emulsifying properties of fish gelatin (FG) at a low concentration (0.5 g 100 g-1) in the high-fat phase (65 g 100 g-1). This study conducted rheology, physicochemical technical and characterization analyses on the emulsions to provide sustainable and innovative approaches for spreadable oils. The addition of WL to FG emulsions improved oxidative stability, emulsion stability and bioactive compounds. The zeta potential (-101 ± 5.62 mV) of 0.58 g 100 g-1 WL-containing emulsion (PE1) was found to be high, whereas particle size (347.6 ± 5.25 nm) and polydispersity index (0.50) were statistically low. It was also found that the addition of WL improved the intermolecular interactions, crystallinity and microstructural properties of the emulsions. All these results were supported by simulating the molecular configuration between FG and WL. The compounds gallic acid, caffeic acid, myricetin, quercetin and resveratrol showed a strong affinity to FG, with free binding energies of -5.50, -5.88, -6.53, -6.68 and -6.66 kcal mol-1, respectively. As a result, WL-supported FG has the potential to be used as an alternative to egg proteins to develop sustainable low-cost spreadable emulsions. © 2024 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Natural Polymer Encapsulated Zeolitic Imidazolate Framework-12 Composite toward Electrochemical Sensing of Antitumor Agent.

Encapsulation of natural polymer pectin (Pec) into a zeolitic imidazolate framework-12 (ZIF-12) matrix via a simple chemical method toward anticancer agent gallic acid (GA) detection is reported in this work. GA, a natural phenol found in many food sources, has gained attention by its biological effects on the human body, such as an antioxidant and anti-inflammatory. Therefore, it is crucial to accurately and rapidly determine the GA level in humans. The encapsulation of Pec inside the ZIF-12 has been successfully confirmed from the physiochemical studies such as XRD, Raman, FTIR, and XPS spectroscopy along with morphological FESEM, BET, and HRTEM characterization. Under optimized conditions, the Pec@ZIF-12 composite exhibits wide linear range of 20 nM-250 μM with a detection limit of 2.2 nM; also, it showed excellent selectivity, stability, and reproducibility. Furthermore, the real sample analysis of food samples including tea, coffee, grape, and pomegranate samples shows exceptional recovery percentage in an unspiked manner. So far, there is little literature for encapsulating proteins, enzymes, metals, etc., that have been reported; here, we successfully encapsulated a natural polymer Pec inside the ZIF-12 cage. This encapsulation significantly enhanced the composite electrochemical performance, which could be seen from the overall results. All of these strongly suggest that the proposed Pec@ZIF-12 composite could be used for miniaturized device fabrication for the evaluation of GA in both home and industrial applications.

Consequences of Gamma Irradiation on Triphala’s Phytochemical Compositions, Microbial Burden and Antioxidant Properties

Triphala, a renowned polyherbal blend comprising three fruits, Phyllanthus emblica Linn., Terminalia chebula Retz., and Terminalia bellirica (Gaertn.) Roxb., in equal proportions, holds a rich historical lineage in both Ayurvedic and Thai traditional medicine. Triphala has been reputed as the indigenous medicine in the Thailand National List of Essential Drugs: List of Herbal Medicinal Products which has been widely used as a natural remedy for relieving cough and phlegm. However, the inherent challenge of microbial contamination in herbal remedies necessitates effective interventions. Gamma () irradiation emerges as a pivotal method to mitigate the microbial burden in medicinal plants, albeit with potential repercussions on their chemical composition and biological properties. This study investigated the impact of -irradiation doses of 5, 10, and 25 kGy exposed to Triphala and the microbial contamination along with antioxidant activity, total phenolic content (TPC), gallic acid (GA), and chebulagic acid (CA) contents. The TPC and antioxidant activity of non-irradiated and irradiated Triphala were determined by using the Folin-Ciocalteu method and DPPH assay, while the GA and CA contents were quantified by HPLC analysis. The results demonstrated the efficacy of radiation doses (5-25 kGy) in diminishing microbial loads without significantly altering TPC or DPPH scavenging activity. Intriguingly, irradiation at 5 and 10 kGy, resulted in a notable increase in GA contents and CA contents (p< 0.05). Thus, the -irradiation emerges as a promising avenue for preserving Triphala quality and antioxidant properties amidst microbial contamination challenges.

Valorization of Pineapple (Ananas comosus) By-Products in Milk Coffee Beverage: Influence on Bioaccessibility of Phenolic Compounds.

The industrial processing of pineapples generates a substantial quantity of by-products, including shell, crown, and core. Bromelain, a proteolytic enzyme found naturally in pineapple, including its by-products, may positively influence the bioaccessibility of phenolics from milk coffee. Therefore, this study aimed to assess how the inclusion of extracts from pineapple by-products, namely shell, crown and core, could impact the bioaccessibility of coffee phenolics when combined with milk. After measuring the proteolytic activity of pineapple by-products, the standardized in vitro digestion model of INFOGEST was employed to evaluate changes in total phenolic content, total antioxidant capacity, and individual phenolic compounds in different coffee formulations. The results showed that incorporating extracts from the crown or core in both black and milk coffee increased the bioaccessibility of total phenolics (from 93 to 114% to 105-129%) and antioxidants (from 54 to 56% to 84-87%), while this effect was not observed for the shell. Moreover, adding core extracts also enhanced the bioaccessibility of caffeoylquinic acids and gallic acid in milk coffee (from 0.72 to 0.85% and 109-155%, respectively). Overall, the findings of this study highlight that bromelain from pineapple core may have a favorable effect on the recovery of phenolic compounds in milk coffee, possibly due to its ability to cleave proteins. These outcomes point out that industrial by-products can be transformed into economic value by being reintroduced into the production process through suitable treatment instead of disposal.

Optimizing the drying conditions of date plum (Diospyros lotus L.) to conserve its phenolic content and antioxidants for preparing a highly bioaccessible polyphenol-rich tea

AbstractDate plum (Diospyros lotus L.) fruits are a good source of bioactive compounds and antioxidants. Drying can increase the shelf life of the fruit and its applications in the food development industry. Optimizing the drying conditions can help to produce prime-quality dried date plum fruits and conserve nutrients including phytochemicals. This study used a two-factor graphics-optimal design to optimize convective drying considering the air velocity and drying temperature of date plum fruits. The independent factors considered included drying temperature (43.78–86.21 °C) and air velocities (0.54–1.96 m/s), and the responses included total phenolic content (TPC), total flavonoid content (TFC), ferric reducing antioxidant power (FRAP), and 2,2-diphenyl-1-picrylhydrazy (DPPH) radical scavenging activity of date plum fruit. The optimized drying conditions (68 °C and 1.75 m/s) resulted in desirable TPC, TFC, FRAP, and DPPH values. The findings indicated that long drying time at low temperatures significantly decreased the phenolics and antioxidants. Date plum tea with different decoction times (5, 10, and 15 min) was prepared from fruits dried at optimum conditions. A decoction time of 5 min resulted in the highest catechin, vanillic, epicatechin, syringic acid, and quercetin-3-glucoside content, which were 2.45 ± 0.04, 11.06 ± 0.11, 22.03 ± 0.11, 12.95 ± 0.08, and 9.37 ± 0.10 mg/L; respectively. In vitro gastrointestinal digestion revealed that the tea product can be a source of highly bioaccessible (> 80%) gallic acid, catechin, vanillic acid, and quercetin-3-glucoside. Applying optimized drying conditions to dehydrate date plum fruit can be useful in preparing a highly bioaccessible polyphenol-rich tea.

Fluorescent Schiff Bases with Phenothiazine and Coumarin Moieties: Synthesis, Characterization, Photophysical, Electrochemical, Computational and Biological Studies.

We present the design and synthesis of four new schiff bases viz., two each of phenothiazine and coumarin derivatives. The structures were proposed based on FT-IR, 1H NMR, 13C NMR and mass spectral data. The photophysical, solvatochromic and electrochemical studies of all the compounds were carried out. Furthermore, theoretical studies such as density functional theory (DFT) were carried out to gain a better understanding of the compounds' intramolecular charge transfer properties and electronic structures. Good agreement was noticed between the HOMO-LUMO energy gap obtained from DFT studies and that calculated from absorption threshold wavelengths. All the compounds showed Stokes shifts in the range of 6345-11,405cm-1. These findings showed that the new schiff bases could be considered as attractive candidates for use in the development of OLEDs, organic electrical devices and optoelectronic devices. Newly synthesized compounds were tested for biological activities. When compared to conventional standards, gallic acid and indomethacin, the schiff bases showed better antioxidant and antiinflammatory activities, respectively.

Camellia Oleifera shells derived nano cellulose crystals conjugated with gallic acid as a sustainable Pickering emulsion stabilizer

Lately, emulsions with low-fat and natural stabilizers are predominant. This study extracted the nano cellulose crystals (NCs) from Camellia Oleifera shells, and their gallic acid (GA) conjugates were synthesized by free-radical grafting. Pickering emulsions were prepared using NCs 1 %, 1.5 %, 2.5 %, and gallic acid conjugates NC-GA1, NC-GA2, and NC-GA3 as stabilizers. The obtained nano cellulose crystals exhibited 18–25 nm, −40.01 ± 2.45 size, and zeta potential, respectively. The contact angle of 83.4° was exhibited by NC-GA3 conjugates. The rheological, interfacial, and microstructural properties and stability of the Pickering emulsion were explored. NC-GA3 displayed the highest absorption content of 79.12 %. Interfacial tension was drastically reduced with increasing GA concentration in NC-GA conjugates. Rheological properties suggested that the low-fat NC-GA emulsions showed a viscoelastic behavior, increased viscosity, gel-like structure, and increased antioxidant properties. Moreover, NC-GA3 displayed reduced droplet size and improved emulsion temperature and storage stability (28 days) against phase separation. POV and TBARS values were reduced with the NC-GA3 (P < 0.05). This work confirmed that grafting phenolic compounds on NCs could enhance bioactive properties, which can be used in developing low-fat functional foods. NC-GA conjugates can potentially fulfill the increasing demand for sustainable, healthy, and low-fat foods.

Fermentation Kinetics, Potability, and Sensory Compounds of Joubidwi: An Ethnic Alcoholic Beverage

Background: Fermentation is a metabolic pathway of the breakdown of carbohydrate molecules like glucose, fructose to ethanol and various volatile and non-volatile congeners. The production of alcoholic beverages from foodstuffs like rice, cereals, fruits, maize etc., is an age-old traditional protocol. In this study, we adapted the traditional protocol of preparation of the ethnic beverage Joubidwi, with a fermentation time of 6 days, i.e., aliquots from the fermented mash (cooked rice) were collected on a daily basis from day 1 to day 6. Objectives: This study aimed to get insight into fermentation kinetics, ethanol content, carbohydrate content, antioxidant activities, bitterness, protein content, pH, colour, and turbidity of the ethnic alcoholic beverage Joubidwi and to identify the sensory compounds and polyphenol molecules. Method: UV-Visible spectroscopic methods, GC-FID and GC-MS analytics methods were used for the estimation and identification of carbohydrates, proteins, ethanol contents, and sensory compounds, respectively. Reverse phase HPLC was adapted for the identification of polyphenol molecules in Joubidwi. Results: A pseudo-first-order fermentation kinetics was observed from day 1 to day 6 fermentation of Joubidwi. Apart from ethanol content (60.48 ± 1.36 mg/mL after 72 h fermentation to 78.80 ± 1.48 mg/mL after 144 h fermentation), the beverage had nutritional values and good antioxidant properties with 22.38 to 35.11% Radical Scavenging Effect (RSE) from 1 to 6th day of fermented Joubidwi. The beverage also contained 12 sensory compounds. Gallic acid was identified as polyphenol in Joubidwi samples. Conclusion: Joubidwi is an alcoholic beverage prepared by Bodos using rice and certain herbs through a traditional fermentation process. The fermentation process follows pseudo-first-order kinetics. The beverage is potable.

Mg-gallate metal-organic framework-based sprayable hydrogel for continuously regulating oxidative stress microenvironment and promoting neurovascular network reconstruction in diabetic wounds

Chronic diabetic wounds are the most common complication for diabetic patients. Due to high oxidative stress levels affecting the entire healing process, treating diabetic wounds remains a challenge. Here, we present a strategy for continuously regulating oxidative stress microenvironment by the catalyst-like magnesium-gallate metal-organic framework (Mg-GA MOF) and developing sprayable hydrogel dressing with sodium alginate/chitosan quaternary ammonium salts to treat diabetic wounds. Chitosan quaternary ammonium salts with antibacterial properties can prevent bacterial infection. The continuous release of gallic acid (GA) effectively eliminates reactive oxygen species (ROS), reduces oxidative stress, and accelerates the polarization of M1-type macrophages to M2-type, shortening the transition between inflammation and proliferative phase and maintaining redox balance. Besides, magnesium ions adjuvant therapy promotes vascular regeneration and neuronal formation by activating the expression of vascular-associated genes. Sprayable hydrogel dressings with antibacterial, antioxidant, and inflammatory regulation rapidly repair diabetic wounds by promoting neurovascular network reconstruction and accelerating re-epithelialization and collagen deposition. This study confirms the feasibility of catalyst-like MOF-contained sprayable hydrogel to regulate the microenvironment continuously and provides guidance for developing the next generation of non-drug diabetes dressings.

Rapid production of abundant transgenic pomegranate (Punica granatum) hairy roots

Pomegranates (Punica granatum) are known for their high levels of health-beneficial compounds that belong to the hydrolyzable tannin (HT) and flavonoid families. However, a significant gap in our understanding exists of the biosynthetic and regulatory genes related to the accumulation of these compounds. To this end, the induction and transformation of hairy roots presents an exciting opportunity to decipher genetically the functions of candidate genes involved in the HT and flavonoid metabolism. This study aims at improving methods for rapid and abundant hairy root production from pomegranate explants by increasing seed germination rates and expediting the attainment of requisite biomass for multifaceted analyses. We found that treating seeds with sandpaper and concentrated sulfuric acid significantly enhanced their germination rates. Interestingly, no correlation between seed-coat texture and seed germination rate was observed among the 20 pomegranate accessions evaluated in this study. Notably, transferring hairy roots from agar plates to soil promoted rapid root biomass growth compared to maintaining them on agar plates. Root biomass as well as levels of gallic acid (a precursor for HT biosynthesis) and punicalagins (the major HTs in roots) across 15 pomegranate accessions were also analyzed. Three of these accessions exhibited higher seed germination percentages along with augmented biomass and elevated punicalagin levels. These pomegranate accessions emerge as promising germplasm for future genetic transformation and functional genomics studies.

Bioassay-guided isolation of anti-inflammatory and antinociceptive metabolites among three Moroccan Juniperus leaves extract supported with in vitro enzyme inhibitory assays.

Herbs of the genus Juniperus (family Cupressaceae) have been commonly used in ancestral folk medicine known as "Al'Araar" for treatment of rheumatism, diabetes, inflammation, pain, and fever. Bioassay-guided isolation of bioactives from medicinal plants is recognized as a potential approach for the discovery of novel drug candidates. In particular, non-addictive painkillers are of special interest among herbal phytochemicals. The current study aimed to assess the safety of J. thurifera, J. phoenicea, and J. oxycedrus aqueous extracts in oral treatments; validating the traditionally reported anti-inflammatory and analgesic effects. Further phytochemical investigations, especially for the most bioactive species, may lead to isolation of bioactive metabolites responsible for such bioactivities supported with in vitro enzyme inhibition. Firstly, the acute toxicity study was investigated following the OECD Guidelines. Then, the antinociceptive, and anti-inflammatory bioactivities were evaluated based on chemical and mechanical trauma assays and investigated their underlying mechanisms. The most active J. thurifera n-butanol fraction was subjected to chromatographic studies for isolating the major anti-inflammatory metabolites. Moreover, several enzymatic assays (e.g., 5-lipoxygenase, protease, elastase, collagenase, and tyrosinase inhibition) were assessed for the crude extracts and isolated compounds. The results showed that acute oral administration of the extracts (300-500 mg/Kg, p.o.) inhibited both mechanically and chemically triggered inflammatory edema in mice (up to 70% in case of J. thurifera) with a dose-dependent antinociceptive (tail flick) and anti-inflammatory pain (formalin assay) activities. This effect was partially mediated by naloxone inhibition of the opioid receptor (2 mg/kg, i.p.). In addition, 3-methoxy gallic acid (1), quercetin (2), kaempferol (3), and ellagic acid (4) were successfully identified being involved most likely in J. thurifera extract bioactivities. Nevertheless, quercetin was found to be the most potent against 5-LOX, tyrosinase, and protease with IC50 of 1.52±0.01, 192.90±6.20, and 399±9.05 μM, respectively. J. thurifera extract with its major metabolites are prospective drug candidates for inflammatory pain supported with inhibition of inflammatory enzymes. Interestingly, antagonism of opioid and non-opioid receptors is potentially involved.

Phlomis crinita Cav. from Algeria: A source of bioactive compounds possessing antioxidant and wound healing activities.

Phlomis crinita Cav. (Lamiaceae), locally known as "El Khayata" or "Kayat El Adjarah", is traditionally used in Algeria for its wound-healing properties. Investigate, for the first time, the phytochemical profile, safety, antioxidant and wound-healing activities of the flowering tops methanolic extract of P. crinita (PCME) collected from Bouira Province in the North of Algeria. Preliminary phytochemical assays were carried out on PCME to quantify the main classes of bioactive compounds, such as total phenols, flavonoids, and tannins. An in-depth LC-DAD-ESI-MS analysis was carried out to elucidate the phytochemical profile of this plant species. Antioxidant activity was investigated by several colorimetric and fluorimetric assays (DPPH, TEAC, FRAP, ORAC, β-carotene bleaching and ferrozine assay). The acute oral toxicity of PCME (2000 mg/kg b.w.) was tested in vivo on Swiss albino mice, whereas the acute dermal toxicity and wound-healing properties of the PCME ointment (1-5% PCMO) were tested in vivo on Wistar albino rats. Biochemical and histological analyses were carried out on biological samples. The phytochemical screening highlighted a high content of phenolic compounds (175.49 ± 0.8 mg of gallic acid equivalents/g of dry extract), mainly flavonoids (82.28 ± 0.44 mg of quercetin equivalents/g of dry extract). Fifty-seven compounds were identified by LC-DAD-ESI-MS analysis, belonging mainly to the class of flavones (32.27%), with luteolin 7-(6''-acetylglucoside) as the most abundant compound and phenolic acids (32.54%), with salvianolic acid C as the most abundant compound. A conspicuous presence of phenylethanoids (15.26%) was also found, of which the major constituent is forsythoside B. PCME showed a strong antioxidant activity with half-inhibitory activity (IC50) ranging from 1.88 to 37.88 μg/mL and a moderate iron chelating activity (IC50 327.44 μg/mL). PCME appears to be safe with Lethal Dose 50 (LD50) ≥ 2000 mg/kg b.w. No mortality or toxicity signs, including any statistically significant changes in body weight gain and relative organs' weight with respect to the control group, were recorded. A significant (p < 0.001) wound contraction was observed in the 5% PCMO-treated group with respect to the untreated and petroleum jelly groups between 8 and 20 days, whereas no statistically significant results were observed at the two lower doses (1 and 2% PCMO). In addition, the 5% PCMO-treated group showed a statistically significant (p < 0.05) wound healing activity with respect to the reference drug-treated group, showing, at the end of the study, the highest wound contraction percentage (88.00 ± 0.16%). PCME was safe and showed strong antioxidant and wound-healing properties, suggesting new interesting pharmaceutical applications for P. crinita based on its traditional use.