P-coumaric Acid Research Articles

Identification and structural characterization of a novel acetyl xylan esterase from Aspergillus oryzae.

Acetyl xylan esterase plays a crucial role in the degradation of xylan, the major plant hemicellulose, by liberating acetic acid from the backbone polysaccharides. Acetyl xylan esterase B from Aspergillus oryzae, designated AoAxeB, was biochemically and structurally investigated. The AoAxeB-encoding gene with a native signal peptide was successfully expressed in Pichia pastoris as an active extracellular protein. The purified recombinant protein had pH and temperature optima of 8.0 and 30 °C, respectively, and was stable up to 35 °C. The optimal substrate for hydrolysis by purified recombinant AoAxeB among a panel of α-naphthyl esters was α-naphthyl acetate. Recombinant AoAxeB catalyzed the release of acetic acid from wheat arabinoxylan. The release of acetic acid from wheat arabinoxylan increased synergistically with xylanase addition. No activity was detected for the methyl esters of ferulic, p-coumaric, caffeic, or sinapic acids. The crystal structures of AoAxeB in the apo and succinate complexes were determined at resolutions of 1.75 and 1.90 Å, respectively. Although AoAxeB has been classified in the Esterase_phb family in the ESTerases and alpha/beta-Hydrolase Enzymes and Relatives (ESTHER) database, its structural features partly resemble those of ferulic acid esterase in the FaeC family. Phylogenetic analysis also indicated that AoAxeB is located between the clades of the two families. Docking analysis provided a plausible binding mode for xylotriose substrates acetylated at the 2- or 3-hydroxy position. This study expands the current knowledge of the structures of acetyl xylan esterases and ferulic acid esterases that are required for complete plant biomass degradation.

Effect of Thermal Treatment on the Retention of Phenolic Compounds, Antioxidant Activity, and Vitamin C of Edible Plants (Mallow Leaf and Garambullo Flower)

Mexico has a wide variety of edible plants, such as mallow and garambullo flowers. Their composition is based on nutrients (carbohydrates, proteins, lipids, vitamins, minerals), as well as phytochemicals, organic or inorganic acids, etc. For the traditional consumption of these plants, they are boiled and then combined with other ingredients. However, after a thermal treatment, their nutritional and sensory qualities are affected. In this work, we study the effect of thermal treatment on the total phenolic compounds (TPC), antioxidant capacity, and vitamin C of two edible plants from Mexico. Both plants had a high amount of TPC as ferulic acid, p-coumaric acid, caffeic acid, quercetin, and apigenin. Both plants had a high amount of TPC as ferulic acid, p-coumaric acid, caffeic acid, quercetin, and apigenin. After thermal treatment, the samples showed a high antioxidant capacity analyzed by 2,2-Diphenyl-1-picrylhydrazyl (DPPH•), 2,2′-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS●+), and ferric reducing antioxidant power (FRAP), associated with a high retention (78 and 89%) of phenolic compounds in the food matrix. Coumarin and ferulic acid were identified in both samples, while apigenin was only found in the mallow leaves and quercetin in the garambullo flowers after thermal treatment. Vitamin C content was higher on mallow leaves compared to garambullo flowers after thermal treatment. Further research is needed to characterize and evaluate the effect in both plants.

Phenolic Acid Profile and In Vitro Antioxidant and Anticholinesterase Activities of Romanian Wild-Grown Acer spp. (Sapindaceae)

This study investigated, for the first time, the phenolic acid profile along with the in vitro antioxidant and acetylcholinesterase (AChE) inhibitory activities of leaves and fruits from five Romanian wild-grown Acer spp. (Sapindaceae): A. campestre L., A. monspessulanum L., A. platanoides L., A. pseudoplatanus L., and A. tataricum L. High-performance liquid chromatography coupled with ultraviolet and mass spectrometry (HPLC/UV/MS) was employed to identify and quantify key phenolic acids, including gallic, caffeic, protocatechuic, chlorogenic, and p-coumaric acids. Total phenolic content (TPC) ranged from 61.48 ± 3.76 to 512.8 ± 20.54 µg gallic acid equivalents (GAE)/mL. Antioxidant activities, assessed through 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) assays, demonstrated the strongest radical scavenging capacities for A. tataricum fruits, with half maximal inhibitory concentration (IC50) values of 10.88 ± 3.39 µg/mL and 10.39 ± 2.86 µg/mL, respectively. The ferric-reducing antioxidant power (FRAP) assay revealed the highest reducing power for A. tataricum fruits (1158 ± 48.98 µmol Fe2+/L) and leaves (1119 ± 164.6 µmol Fe2+/L). AChE inhibition was only found in A. tataricum, with an IC50 of 7.91 ± 7.50 mg/mL for leaves and 15.95 ± 8.52 mg/mL for fruits. These results highlight A. tataricum as a promising source of natural antioxidants and neuroprotective agents.

Propolis Hydroalcoholic Extracts: Biochemical Characterization and Antifungal Efficacy

The present study investigated the antifungal potential of hydroalcoholic extracts of propolis against the causal agent of grey mould, Botrytis cinerea, by in vitro and in vivo assays. Five different propolis from different Italian regions were subjected to hydroalcoholic extraction using different ethanol concentrations and extraction methods. The preliminary bio-assay showed significant inhibitory effects on B. cinerea mycelial growth of propolis extracts obtained using 90% ethanol and subjected to sonication. The calculation of EC50 values, based on the demonstrated efficacy of non-volatile and volatile metabolites of propolis extracts, was useful to understand the main fraction involved in the antifungal activity of the samples and to perform the in vivo assay on grape and blueberry fruits. Three of the propolis extracts showed a high amount of genistein. Conversely, the other two propolis showed a fair amount of apigenin, caffeic acid, chrysin, ferulic acid, kaempferol, luteolin, p-coumaric acid and quercetin. From the volatile analysis of propolis, the main compounds detected were α-cadinol, α-eudesmol, calamenene, cadinol, benzyl acetate, benzyl benzoate and benzyl alcohol. The results suggest that propolis extracts have potential as an effective postharvest antifungal treatment, with varying degrees of efficacy depending on the extraction method and the type of propolis metabolites.

A Therapeutic Approach of Chitosan-loaded p-Coumaric Acid Nanoparticles to Alleviate Diabetic Nephropathy in Wister Rats.

This study evaluated the renoprotective effects of p-Coumaric acid nanoparticles (PCNPs) in nephropathic rats. Six groups of male Albino Wistar rats were randomly assigned. Group 1 was the control, while Group 2 received 45 mg/kg of streptozotocin (STZ) to induce diabetic nephropathy. Groups 3, 4, and 5 received STZ (45 mg/kg) along with PCNPs at doses of 20, 40, and 80 mg/kg, respectively. Group 6 received 80 mg/kg of PCNPs without STZ. Body weight, blood glucose, insulin, hemoglobin (Hb), and glycosylated hemoglobin (HbA1c) levels were measured. Blood urea, serum creatinine, kidney antioxidant enzymes, and lipid peroxidation levels were also analyzed. Histological and immunohistochemical studies of kidney tissues were performed. PCNPs (80 mg/kg) significantly reduced serum glucose, creatinine, and urea levels while increasing insulin levels and antioxidant activity in the kidneys. Histological analysis revealed that nephropathic rats exhibited cellular damage, including shrinkage of Bowman's capsule and lesions in the kidneys, along with degeneration in the Islets of Langerhans in the pancreas. PCNPs treatment restored these morphological alterations in the pancreas, liver, and kidneys to near-normal. Furthermore, nephropathic rats had elevated IL-6 and TNF-α expression in the renal tubules and glomeruli, which was reduced following PCNPs treatment. The findings suggest that PCNPs exhibit antihyperglycemic, antioxidant, antiglycation, and renoprotective effects in STZ-induced diabetic nephropathy.

Automated adjustment of metabolic niches enables the control of natural and engineered microbial co-cultures.

Much attention has focused on understanding microbial interactions leading to stable co-cultures. In this work, substrate pulsing was performed to promote better control of the metabolic niches (MNs) corresponding to each species, leading to the continuous co-cultivation of diverse microbial organisms. We used a cell-machine interface, which allows adjustment of the temporal profile of two MNs according to a rhythm, ensuring the successive growth of two species, in our case, a yeast and a bacterium. The resulting approach, called 'automated adjustment of metabolic niches' (AAMN), was effective for stabilizing both cooperative and competitive co-cultures. AAMN can be considered an enabling technology for the deployment of co-cultures in bioprocesses, demonstrated here based on the continuous bioproduction of p-coumaric acid. The data accumulated suggest that AAMN could be used not only for a wider range of biological systems, but also to gain fundamental insights into microbial interaction mechanisms.

Integrating Physiology, Transcriptome, and Metabolomics Reveals the Potential Mechanism of Nitric Oxide Concentration-Dependent Regulation of Embryo Germination in Sorbus pohuashanensis

Nitric oxide (NO) breaks a seed’s dormancy and stimulates germination by signaling. However, the key physiological metabolic pathways and molecular regulatory mechanisms are still unclear. Therefore, this study used physiological, transcriptomic, and metabolomics methods to analyze the key genes and metabolites involved in the NO regulation of plant embryo germination and their potential regulatory mechanisms. The physiological analysis results indicate that the appropriate concentration of NO increased the content of NO and hydrogen peroxide (H2O2) in cells, stimulated the synthesis of ethylene and jasmonic acid (JA), induced a decrease in abscisic acid (ABA) content, antagonistic to the gibberellin (GA3) effect, and promoted embryo germination and subsequent seedling growth. However, the high concentrations of NO caused excessive accumulation of H2O2, destroyed the reactive oxygen species (ROS) balance, and inhibited embryo germination and seedling growth. The combined analysis of transcriptomics and metabolomics showed that the genes related to phenylpropanoid (phenylalanine ammonia-lyase, trans-cinnamate 4-monooxygenase, ferulate-5-hydroxylase, coniferyl-alcohol glucosyltransferase), and flavonoid synthesis (10 genes such as CHS) were significantly up-regulated during embryo germination. The high concentration of exogenous NO inhibited embryo germination by up-regulating the expression of 4-coumaric acid coenzyme A ligase (4CL) and negatively regulating the expression of flavonoid synthesis genes. This suggests that NO concentration-dependently regulates phenylpropanoid and flavonoid biosynthesis, thereby affecting ROS metabolism and hormone levels, and ultimately regulates the dormancy and germination of Sorbus pohuashanensis embryos.

Evaluation of the antibacterial and inhibitory activity of the NorA and TetK efflux pumps of Staphylococcus aureus by p-coumaric acid.

The NorA and TetK efflux pumps mediate resistance to fluoroquinolone and tetracycline antibiotics by actively extruding these compounds and reducing their intracellular concentrations. Consequently, intense research has focused on inhibiting these efflux mechanisms using antimicrobial agents derived from natural or synthetic sources. This study used Fourier transform infrared spectroscopy (ATR-FTIR) to analyze the various functional groups present in p-coumaric acid. We also investigated the antibacterial activity of p-coumaric acid on strains of Staphylococcus aureus carrying the NorA and TetK efflux pumps, as well as the compound's ability to increase the fluorescence of ethidium bromide (EtBr) and Sytox Green. In addition, the interactions of this compound with NorA were analyzed using molecular docking, and its pharmacokinetic properties were evaluated using ADMET (Absorption, Distribution, Metabolism, Excretion and Toxicity) modeling. The results revealed that p-coumaric acid exhibited no direct antibacterial activity against the tested Staphylococcus aureus strains. However, significant reductions in the minimum inhibitory concentrations (MICs) of norfloxacin and EtBr, both used as NorA substrates, were observed when combined with p-coumaric acid. It was also observed that p-coumaric acid increased the fluorescence emission of EtBr and Sytox Green in strains 1199 and 1199B, suggesting the inhibition of the efflux mechanism and enhanced membrane permeabilization in S. aureus. The in silico analysis demonstrated that p-coumaric acid exhibits a favorable binding energy with NorA, comparable to that of chlorpromazine. These results position p-coumaric acid as a promising antibiotic adjuvant and efflux inhibitor in strains harboring NorA.

Identification of chemical constituents and absorbed components in rat serum of Ipomoea muricata (L.) Jacq. by UPLC-Q-exactive-MS

Ipomoea muricata (L.) Jacq. has anti-inflammatory, analgesic, haemostatic and antioxidant effects, but its pharmacological components are still unclear. This study comprehensively identified chemical constituents and absorbed components of I. muricata by UPLC-Q-Exactive-MS. The column temperature was 35 °C, and the injection volume was 2 μl. The mobile phase using a 0.1% formic acid as phase A and 0.1% formic acid of solution in methanol as phase B was adopted in positive ion mode. And the mobile phase using a 10 mM formic ammonia solution as phase A and 10 mM formic ammonia of solution in 95% methanol as phase B was adopted in negative ion mode. In this study, we identified 25 compounds from the alcoholic extract of I. muricata, including 11 organic acids, 5 coumarins, 2 flavonoids, 7 alkaloids and nitrogen-containing compounds. Nine absorbed components were identified from the serum of rats after administration, including adenosine, p-hydroxybenzoic acid, chlorogenic acid, dextroquinic acid, shikimic acid, caffeic acid, p-hydroxycinnamic acid, chrysin, and umbelliferone. UPLC-Q-Exactive-MS technique can quickly identify the chemical constituents and absorbed components in rat serum of I. muricata, and provide scientific basis for clarifying the pharmacodynamic material basis and optimising the quality control index of I. muricata.

Metabolic engineering of Escherichia coli for enhanced production of p-coumaric acid via L-phenylalanine biosynthesis pathway.

p-Coumaric acid (p-CA), an invaluable phytochemical, has novel bioactivities, including antiproliferative, anxiolytic, and neuroprotective effects, and is the main precursor of various flavonoids, such as caffeic acid, naringenin, and resveratrol. Herein, we report the engineering of Escherichia coli for de novo production of p-CA via the PAL-C4H pathway. As the base strain, we used the E. coli H-02 strain, which was previously engineered for sufficient supplementation of L-phenylalanine, the main precursor of p-CA. For the bioconversion of L-Phe to p-CA, we constructed and optimized an expression system for phenylalanine ammonia lyase (SmPAL), codon-optimized cinnamate 4-hydroxylase (AtC4H), and its redox partner, cytochrome P450 reductase (AtCPR1). We confirmed that the engineered cell showed higher production of p-CA at 30°C and the addition of 0.5mM 5-aminolevulinic acid could increase the production titer further. Subsequently, the main pathways of acetic acid (poxB and pta-ackA) were eliminated to reduce its accumulation and restore cell growth. Next, to increase the available pool of cofactor (NADPH), the co-expression system of the zwf gene in the pentose phosphate pathway (PPP) was integrated into genome and the expression level was optimized with synthetic promoters. Finally, by optimizing fed-batch culture in a 5 L-scale bioreactor, the engineered strain achieved 1.5g/L p-CA with a productivity of 31.8mg/L/h.

Impact of a coating of pectin and Cinnamomum Zeylanicum essential oil on the postharvest quality of strawberries packaged under different modified atmosphere conditions

Extending shelf life during storage and distribution has always represented a significant challenge in the value chain of strawberries and other perishable fruits. Thus, the combined effect of two Modified atmosphere packaging (MAP) techniques and pectin-based coating with Cinnamomum zeylanicum essential oil (CEO) was investigated on preservation of ‘Palmaritas’ variety. Strawberries were exposed to four treatments during 12 days of storage: Normal atmosphere without coating (control), normal atmosphere and a pectin-based coating with essential oil (NEC), modified atmosphere of 2.5% O2 + 15% CO2 + 82.5% N2, a pectin coating with essential oil (EC15CO2) and a modified atmosphere of 2.5% O2 + 20% CO2 + 77.5% N2, and a pectin coating with essential oil (EC20CO2). The studied parameters were weight loss, decay rate, firmness, color, total soluble solids (TSS), total phenolic content (TPC), total anthocyanin content (TAC), total flavonoid content (TFC), antioxidant capacity (AC), and phenolic and organic compounds. Results showed that a pectin-based coating with 0.17% Cinnamomum zeylanicum essential oil in a CO2-enriched atmosphere guaranteed significant preservation of several quality attributes of strawberries during storage at 6° C, especially firmness, color, TSS, TPC, TAC and TFC. Moreover, our total phenolic compounds measurement revealed an increase in gallic acid, trans-ferulic acid, kaempferol, and malvidin 3-O-glucoside, coupled with notably less degradation of caffeic acid, p-coumaric acid, and quercetin than in the control sample. Applying a CEO coating and storage in MAP effectively preserved organic acids, particularly oxalic acid and malic acid, while slowing down the degradation of vitamin C and citric acid. Thus, bioactive coating coupled with MAP presents a promising alternative for fruit storage, allowing a significant shelf-life extension while maintaining and stabilizing the principal quality attributes of strawberries.

How Phenolic Compounds Profile and Antioxidant Activity Depend on Botanical Origin of Honey-A Case of Polish Varietal Honeys.

Honey contains natural biologically active compounds, and its preventive and healing properties are primarily linked to its antioxidant activity. The antioxidant properties of honey can be related to the botanical origin and content of phenolic compounds. We tested 84 honey samples from Poland, representing eight honey varieties: acacia, phacelia, buckwheat, linden, rapeseed, heather, goldenrod, and honeydew. High-performance liquid chromatography with photodiode-array detection (HPLC-DAD) was used to determine the phenolic compound composition of honey extracts. Total phenolic compounds (TPC) and DPPH radical-scavenging activity were also evaluated. We detected vanillin aldehyde, vanillic acid, caffeic acid, p-coumaric acid, and trans-ferulic acid, as well as flavonoid pinocembrin, in all honey varieties. The results of our study showed that honeys with high antioxidant activity were characterized by significantly higher total phenolic compounds content. Neither clustering method nor principal component analysis (PCA) showed clear separation of each honey variety, possibly due to high intra-variety diversities. We suppose that the variability of qualitative and quantitative phenolic compound composition within honey varieties may result from the region of origin, secondary nectar sources, and the time of harvest.

Polyphenolic compounds in Sauvignon Blanc - from grapes to wine

At harvest, the metabolic composition of wine grapes reflects the accumulated effect of the environmental conditions, the stresses endured, and viticultural manipulations applied during the growing season. The role of the winemaker is to extract and nurture this “metabolite potential” throughout the winemaking process. However, it is typically difficult to relate this grape potential to that of the eventual wines. In this study, a holistic view of Sauvignon Blanc grape and wine polyphenolic compounds was attempted by measuring these compounds from different matrices, from ripe grape tissues up to the final wine, including the submatrices, such as juice, pomace and sediment. Sauvignon Blanc vines from one vineyard block were manipulated to yield berries with distinctly different phenolic potentials by creating a high light (HL) or a low light (LL) microclimate in the fruiting zone of the canopy during the growing season. The analyses of the HL and LL berries and wines, as well as concomitant analyses of the phenolic compounds in the submatrices, allowed their tracing as they were (i) transferred from one matrix to another, (ii) lost as waste products, or (iii) affected by different winemaking practices (skin contact and/or fermentation in contact with the juice sediment) implemented in the experimental design. In berries, flavonols showed the largest increase due to sun exposure (HL treatment) but were absent from the juice samples at all juice processing stages. They were however detected in the juice sediment, together with high concentrations of organic acids and sugars. Juice processing was noted for dramatic fluctuations in metabolite concentrations suggesting intense metabolic activity in this pre-fermentation matrix. Both skin contact and sediment contact treatments delivered wines with higher concentrations of coutaric acid (the ester formed from coumaric and tartaric acid) and the flavanol catechin while epicatechin concentrations was unaffected. The higher catechin concentrations did not lead to increased perceived bitterness in the wines, except in the sediment contact treatments. The total phenolic compound concentration of wine from LL (low phenolic potential) grapes was comparable to wine from HL (high phenolic potential) grapes, when skin contact, or sediment contact treatments were employed. From a sensory perspective, the sediment contact decreased the fruity aromas of Sauvignon Blanc, while the skin contact treatment enhanced the sensorial properties of the wines made from the LL grapes, allowing increased extraction from the skin-accumulated impact compounds.

Bioactive Compounds from Propolis on Bone Homeostasis: A Narrative Review.

This narrative review explores the potential effects of Propolis and its bioactive compounds on bone health. Propolis, a resinous product collected by bees, is renowned for its antimicrobial, anti-inflammatory, and antioxidant properties. Recent research emphasizes its positive role in osteogenesis, primarily through the modulation of osteoclast and osteoblast activity via molecular pathways. Key mechanisms include reducing inflammatory cytokines, protecting against oxidative stress, and upregulating growth factor essential for bone formation. While compounds such as Caffeic Acid Phenethyl Ester, Apigenin, Quercetin, and Ferulic Acid have been well-documented, emerging evidence points to the significant roles of less-studied compounds like Pinocembrin, Kaempferol, p-Coumaric acid, and Galangin. This review synthesizes the current literature, focusing on the mechanisms by which these bioactive compounds influence osteogenesis. Firstly, it explores the techniques for characterizing bioactive compounds presented in propolis, the chemogeographic variations in its composition, and the effects of both crude extracts and isolated compounds on bone tissue, offering a comprehensive analysis of recent findings across different experimental models. Further, it discusses the effects of Propolis compounds on bone health. In summary, these compounds modulate signaling pathways, including nuclear factor kappa beta, wingless-related integration site, mitogen-activated protein kinase, vascular endothelial growth factor, and reactive oxygen species. These pathways influence the receptor activator of nuclear factor kappa-β/receptor activator of nuclear factor kappa-β ligand/osteoprotegerin system, fostering bone cell differentiation. This regulation mitigates excessive osteoclast formation, stimulates osteoblast activity, and ultimately contributes to the restoration of bone homeostasis by maintaining a balanced bone remodeling process.

Changes in the Content of Dietary Fiber, Flavonoids, and Phenolic Acids in the Morphological Parts of Fagopyrum tataricum (L.) Gaertn Under Drought Stress.

Tartary buckwheat is a plant recognized for its resistance to various environmental stresses. Due to its valuable source of phenolic compounds, Fagopyrum tataricum is also characterized as a medicinal plant; therefore, the aim of this study was to investigate the drought stress for the levels of phenolic compounds in the morphological parts of the plant. This experiment was conducted in 7 L pots under laboratory conditions. Phenolic compounds were identified using a UHPLC-MS chromatography system. Antioxidant activity was assessed using well-known methods, including the DPPH scavenging activity and ferrous ion chelating activity. In Tartary buckwheat leaves, stems, seeds, and husks, 57 phenolic compounds were identified, with a predominance of quercetin 3-rutinoside, quercetin, kaempferol-3-rutinoside, kaempferol, and derivatives of coumaric acid. It was observed that the Tartary buckwheat samples subjected to drought stress exhibited a slight decrease in the majority of individual phenolic compounds. The measurement of biological parameters indicated that plant regeneration after drought stress demonstrated a rapid recovery, which can be a positive response to the progression of climate changes.

Cinnamic Acid Compounds (p-Coumaric, Ferulic, and p-Methoxycinnamic Acid) as Effective Antibacterial Agents Against Colistin-Resistant Acinetobacter baumannii.

Colistin-resistant Acinetobacter baumannii (COLR-Ab) is an opportunistic pathogen commonly associated with nosocomial infections, and it is difficult to treat with current antibiotics. Therefore, new antimicrobial agents need to be developed for treatment. Based on this information, we investigated the antimicrobial, antibiofilm, and combination activities of p-coumaric acid (p-CA), ferulic acid (FA), and p-methoxycinnamic acid (p-MCA) against five COLR-Ab isolates. p-CA, FA, and p-MCA exhibited antimicrobial activity against COLR-Ab isolates, with minimum inhibitory concentration (MIC) values in the range of 256-128 µg/mL, 1024-512 µg/mL, and 512-128 µg/mL, respectively. The combination effects of the compounds with colistin (COL) were evaluated using a checkerboard synergy test. The combinations exhibited a synergistic effect and caused a 128- to 16-fold decrease in COL MIC values. In addition, the biofilm production capacities of the COLR-Ab isolates and the antibiofilm activities of the compounds were determined using the microtitre plate-based crystal violet (CV) technique. The compounds showed effective antibiofilm activity against strong and moderate biofilm-producing isolates, inhibiting biofilm formation by 77.5% and 19.7%. Spectrometric measurements were used to examine the effect of compounds on membrane permeability; 1.9-, 1.66-, and 1.34-fold increases in absorbance values were observed at MIC concentrations of p-CA, FA, and p-MCA, respectively. Furthermore, morphological changes caused by the compounds in the isolate were observed using scanning electron microscopy (SEM) micrographs. According to the WST assay, the compounds did not show any statistically significant cytotoxic effect on the cells (p > 0.05). These findings indicate that p-CA, FA, and p-MCA may be potential new alternative candidates against resistant A. baumannii.

Screening of Leuconostoc mesenteroides strains suitable for kimoto-style brewing of sake with high antioxidant capacity.

Sake brewed using the kimoto-style exhibits high antioxidant capacity and is expected to inhibit the deterioration of sake quality due to oxidation. However, the antioxidant capacity of the added lactic acid bacteria has not been explored. We aimed to screen the lactic acid bacterium, Leuconostoc mesenteroides, with excellent brewing and antioxidant capacity, to develop sake with high antioxidant capacity. Three Le. mesenteroides strains (19-2, 19-5, and 19-23) were selected from eight screened strains based on their alcohol intolerance, growth performance in koji extract, aroma compound, and low biogenic amine production. Among these, Le. mesenteroides 19-23 exhibited a significantly higher hydrophilic-oxygen radical absorbance capacity (H-ORAC) in the culture medium than the control strain, Le. mesenteroides NBRC102481. In a medium-scale sake brewing test, sake prepared with Le. mesenteroides 19-23 had a significantly higher H-ORAC value than that prepared using the sokujo-style (without Le. mesenteroides). Additionally, metabolome analysis using capillary electrophoresis time-of-flight mass spectrometry identified ferulic acid, p-coumaric acid, 3-hydroxyanthranilic acid (3-HAA), and 6,8-thioctic acid in the main mash. High-performance liquid chromatography-based quantification revealed that antioxidants in the unrefined filtrates of the main mash prepared using the kimoto-style tended to be more abundant than in those prepared using the sokujo-style. Specifically, 3-HAA and ferulic acid were more concentrated in some unrefined filtrates of the kimoto-style than sokujo-style. In conclusion, the screened Le. mesenteroides strain demonstrated potential for brewing sake with high antioxidant capacity using the kimoto-style, offering a promising method for antioxidant-rich sake production.

Artificial intelligence-driven identification and mechanistic exploration of synergistic anti-breast cancer compound combinations from Prunella vulgaris L.-Taraxacum mongolicum Hand.-Mazz. herb pair.

The Prunella vulgaris L. (PVL) and Taraxacum mongolicum Hand.-Mazz. (TH) herb pair, which is commonly used in traditional Chinese medicine (TCM), has been applied for the treatment of breast cancer. Although its efficacy is validated, the synergistic anti-breast cancer compound combinations within this herb pair and their underlying mechanisms of action remain unclear. This study aimed to identify and validate synergistic anti-breast cancer compound combinations within the PVL-TH pair using large-scale biomedical data, artificial intelligence and experimental methods. The first step was to investigate the anti-breast cancer effects of various PVL and TH extracts using in vitro cellular assays to identify the most effective superior extracts. These superior extracts were subjected to liquid chromatography-mass spectrometry (LC-MS) analysis to identify their constituent compounds. A deep learning-based prediction model, DeepMDS, was applied to predict synergistic anti-breast cancer multi-compound combinations. These predicted combinations were experimentally validated for their anti-breast cancer effects at actual content ratios found in the extracts. Preliminary bioinformatics analyses were conducted to explore the mechanisms of action of these superior combinations. We also compared the anti-breast cancer effects of superior extracts from different geographical origins and analyzed the contents of compounds to assess their representation of the anti-tumor effect of the corresponding TCM. The results revealed that LC-MS analysis identified 27 and 21 compounds in the superior extracts (50% ethanol extracts) of PVL and TH, respectively. Based on these compounds, DeepMDS model predicted synergistic anti-breast cancer compound combinations such as F973 (caffeic acid, rosmarinic acid, p-coumaric acid, and esculetin), T271 (chlorogenic acid, cichoric acid, and caffeic acid), and T1685 (chlorogenic acid, rosmarinic acid, and scopoletin) from single PVL, single TH and PVL-TH herb pair, respectively. These combinations, at their actual concentrations in extracts, demonstrated superior anti-breast cancer activity compared to the corresponding extracts. The bioinformatics analysis revealed that these compounds could regulate tumor-related pathways synergistically, inhibiting tumor cell growth, inducing cell apoptosis, and blocking cell cycle progression. Furthermore, the concentration ratio and total content of compounds in F973 and T271 were closely associated with their anti-breast cancer effects in extracts from various geographical origins. The compound combination T1685 could represent the synergistic anti-breast cancer effects of the PVL-TH pair. This study provides insights into exploring the representative synergistic anti-breast cancer compound combinations within the complex TCM.

Advanced Glycation End-Product-Modified Heat Shock Protein 90 May Be Associated with Urinary Stones.

Urinary stones (urolithiasis) have been categorized as kidney stones (renal calculus), ureteric stones (ureteral calculus and ureterolith), bladder stones (bladder calculus), and urethral stones (urethral calculus); however, the mechanisms underlying their promotion and related injuries in glomerular and tubular cells remain unclear. Although lifestyle-related diseases (LSRDs) such as hyperglycemia, type 2 diabetic mellitus, non-alcoholic fatty liver disease/non-alcoholic steatohepatitis, and cardiovascular disease are risk factors for urolithiasis, the underlying mechanisms remain unclear. Recently, heat shock protein 90 (HSP90) on the membrane of HK-2 human proximal tubular epithelium cells has been associated with the adhesion of urinary stones and cytotoxicity. Further, HSP90 in human pancreatic and breast cells can be modified by various advanced glycation end-products (AGEs), thus affecting their function. Hypothesis 1: We hypothesized that HSP90s on/in human proximal tubular epithelium cells can be modified by various types of AGEs, and that they may affect their functions and it may be a key to reveal that LSRDs are associated with urolithiasis. Hypothesis 2: We considered the possibility that Japanese traditional medicines for urolithiasis may inhibit AGE generation. Of Choreito and Urocalun (the extract of Quercus salicina Blume/Quercus stenophylla Makino) used in the clinic, Choreito is a Kampo medicine, while Urocalun is a characteristic Japanese traditional medicine. As Urocalun contains quercetin, hesperidin, and p-hydroxy cinnamic acid, which can inhibit AGE generation, we hypothesized that Urocalun may inhibit the generation of AGE-modified HSP90s in human proximal tubular epithelium cells.

Optimization of UAE-NADES green extraction of bioactive compounds from chickpea (Cicer arietinum L.) sprouts using simplex lattice mixture design methodology

In the present study, a statistical tool called the simplex lattice mixture design method was used to create a new formulation of Natural Deep Eutectic Solvent (NADES) which is derived from a combination of three compounds (citric acid, glycerol, and water) to extract bioactive compounds from chickpea (Cicer arietinum L.) sprouts. The mixture (natural deep eutectic solvent) was formulated by combining three solvents including citric acid, glycerol, and water. The extraction was performed in a sonication bath for 30 min. The simultaneous optimization was performed to obtain the highest total polyphenol content (TPC), total flavonoid content (TFC) and antioxidants activity. The highest values of total polyphenol content (TPC), total flavonoid content (TFC) and antioxidant activity were 128.0 ± 0.2 mg GAE/100 g, 38.61 ± 0.03 mg CE/100 g and 2117 ± 1.8 µmol TE/100 g respectively. HPLC-DAD of the optimized extract was utilized for quantification of polyphenol compounds showing catechin as the main compound followed by chlorogenic acid, epicatechin, syringic acid, rutin, gallic acid, kaempferol 3-glucoside, ferulic acid, and coumaric acid. These findings may represent a significant advancement in the management of phenolic compound extraction for targeted uses, such as serving as alternatives to traditional antioxidants primarily employed in the food industry to improve nutritional quality. Furthermore, our research has shown that mixture designs are an efficient and useful method for structuring and optimizing experimental parameters to achieve the most accurate results with the minimum number of experiments.