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Practical ApplicationIn recent years, red‐fleshed apple has attracted a lot of attention due to its pleasant appearance, taste, and being a valuable source of anthocyanins. Generally, different types of this apple are known as ornamental apple plants that are rarely found in various regions around the world like New Zealand, Kazakhstan, Kyrgyzstan, Australia, China, and Turkey. This study focused on the accumulation and changes of the bioactive and aroma‐related volatile components at different production stages of a red‐fleshed apple vinegar. The aim of this study is to produce an anthocyanin‐ and vitamin C‐rich vinegar (NAV) from red‐fleshed Niedzwetzky's apple for the first time. According to the results, in this vinegar, it was determined high concentrations of total phenolic content (904.8 mg‐GAE/mL), total flavonoid content (0.25 mg‐EGCE/mL), vitamin C content (3024.3 mg‐AAE/mL), and 1,1‐diphenyl‐2‐picrylhydrazyl (8.33 mmol‐TE/mL), as well as FRAP (13.57 mmol‐ISE/mL), respectively. Moreover, total anthocyanin content value was found to be 6.46 mg/g. One of the components that provide the specified functional effect in this vinegar is chlorogenic acid, which constitutes the main phenolic compound, and the other one is cyanidin‐3‐glucoside, which is the major anthocyanin in this vinegar. The main volatile components associated with the aroma of vinegar are ‐acetic acid, ‐phenylacetic acid (5067.7 µg/100 mL), ‐phenethyl alcohol (3096.1 µg/100 mL), and ‐nonanoic acid (2939.0 µg/100 mL) compounds. Consequently, it is recommended to expand the range of Niedzwetzky's apples in the production and consumption of food products such as functional vinegar. Thus, a new vinegar with high functional properties will be introduced to the worldwide food industry.In this study, vinegar was produced from the small and dark‐red‐flesh‐colored fruits of Niedzwetzky's apple tree, which is known in a small part of the world and is generally used as an ornamental plant due to its red‐pink flowers. It has been determined that the produced vinegar has high bioavailability due to the apples with red inner flesh and high anthocyanin content and has a desirable aromatic content. Thus, a food product with high functional quality and desirable aroma, accessible to consumers all over the world, has been produced from a little‐known fruit. [ABSTRACT FROM AUTHOR]

Elephantopus tomentosus (ET) Linn. was reported to be an anti-tumor plant. However, the chemical composition of ET and its anti-tumor compounds and potential mechanisms still unclear. In this paper, UPLC-Q-TOF–MS/MS was firstly used to identified the ingredients in ET and UPLC was used to determine the main compounds of ET. Network pharmacology was applied to predict the potential mechanisms of anti-liver cancer. Anti-tumor nuclear activate compounds and targets of ET were obtained and the anti-liver cancer effect was validated on HepG2. Finally, Molecule docking, RT-qPCR, and western blotting were used for verification of the relationship between nuclear activate compounds and nuclear targets and the potential anti-cancer mechanisms. The result showed that 42 compounds were identified in ET, which consisted of sesquiterpene lactones, flavonoids, and phenylpropanoid compounds. Scabertopin (ST), chlorogenic acid, Isochlorogenic acid B, Isochlorogenic acid A and Isochlorogenic acid C were identified as main compounds and were determined as 0.426%, 0.457%, 0.159%, 0.701%, and 0.103% respectively. 24 compounds showed high pharmacokinetics and good drug-likeness. 520 overlapping targets of the ET compounds and liver cancer were collected. The targets were used for KEGG and GO analysis. GO enrichment analysis suggested that the targets of 24 active compound closed related to promote apoptosis, inhibit proliferation, and regulate oxidative levels. KEGG enrichment analysis suggested that pathway in cancer was enriched most and p38 MAPK/p53 signaling pathway, which closely related to promoting apoptosis and inhibiting proliferation. Compounds-targets analysis based on the parameter of Betweenness, Closeness, Information, Eigenvector, Degree, and component content indicated that ST was the nucleus anti-tumor active compound of ET. HepG2 was first used to validated the anti-tumor effect of ST and the result showed that ST significantly inhibited HepG2 proliferation with a low IC50 less than 5 μM. Nucleus active compound targets, including TP53, CASP3, BCL2, EGFR, TNF-a, IL-1β, and IL-6 were enriched based on degree value of PPI analysis. Molecule docking suggested that ST showed a good combination to TGFBR1 with the combination energy less than − 5 kcal/mol. RT-qPCR result also suggested that ST significantly medicated the mRNA expression level of TP53, CASP3, BCL2, EGFR, TNF-a, IL-1β, and IL-6. Protein expression of p-p38/p38 and p-p53/p53 notable increased by ST treatment. In conclude, combining with UPLC-Q-TOF–MS/MS qualitative analysis, UPLC quantitative analysis, network pharmacology analysis, molecule docking, and in vitro experiments on HepG2, we suggest that ST is an anti-tumor ingredient of ET, which may target to TGFBR1 and promote apoptosis and inhibited proliferation of HepG2 by activating p38 MAPK/p53 signaling pathway. ST can be regarded as a quality marker of ET. [ABSTRACT FROM AUTHOR]

In this study, the fatty acid content, phenolic compounds, and antimicrobial activity of seed oils from Ege Özbaş (EO), Sarıbağlar 407 (S407) and Akhisar 98 (A98) tobacco plants grown in the Aegean region were investigated. The ozonation process altered fatty acid and phenolic compound compositions and affected antimicrobial activities against Gram-positive and Gram-negative bacteria in the oils. Tobacco seed oils (TSOs) were extracted via cold pressing (CP) and Soxhlet extraction (SE). Fatty acids and phenolic compounds were analyzed using GC-FID and LC-MS/MS, respectively. Non-ozonated oils averaged 73% linoleic acid, 13% oleic acid, 9% palmitic acid, and 3% stearic acid, while ozonated oils contained 41% linoleic acid, 18% oleic acid, 22% palmitic acid, and 11% stearic acid. The presence of 33 phenolic compounds was investigated and 22 common phenolic compounds were detected in both ozonated and non-ozonated TSOs, including gallic acid, protocatechuic acid, chlorogenic acid, 3-hydroxybenzoic acid, (-)-epicatechin, 2,5-dihydroxybenzoic acid, caffeic acid, verbascoside, luteolin-7-glucoside, hesperidin, and rosmarinic acid. The increased antimicrobial activity of fatty acids and phenolic compounds following ozonation suggests the potential for developing creams to treat skin diseases and wounds caused by antibiotic-resistant pathogens, and to reduce the effects of aging skin wrinkles. [ABSTRACT FROM AUTHOR]

Background: Sea buckthorn (Hippophae rhamnoides), a deciduous species plant, is widely distributed around the globe, and native to the cold-temperate regions of Europe and Asia. This medicinal herb contains several bioactive constituents including chlorogenic acid. The conventional methods used for the extraction of phenolic antioxidants from natural herbs often result in low yields, high toxicity, and pose environmental hazards limiting their effectiveness and scalability. Therefore, green extraction techniques using deep eutectic solvents, composed of natural, non-toxic, and biodegradable components were applied for extraction of chlorogenic acid from sea buckthorn weed. Fourteen deep eutectic solvent mixtures were prepared and evaluated for extraction yield of chlorogenic acid. Parameters such as hydrogen bond donor-to-hydrogen bond acceptor ratio, liquid-to-solid ratio, shaking speed, and shaking time were optimized for the best mixture. Results: The combination of lactic acid and maltose (1:1) was found to give best extraction yield using response surface methodology. The deep eutectic solvent system under optimum conditions produced 12.2 g/100 g of crude extract sea buckthorn containing 174.7 mg gallic acid equivalents (mg GA)/g) of extract. Moreover, the optimized extract exhibited appreciable radical scavenging capacity (91%), trolox equivalent antioxidant capacity (11.2% of extract), and inhibition of peroxide in linoleic acid (80.6%). High-performance liquid chromatography-based characterization revealed the extracts contained chlorogenic acid (20.1 mg/g of extract) as the major constituent. Conclusions: In summary, the adoption of DES for the extraction of bioactive phenolic constituents from sea buckthorn offers multiple benefits, including economic efficiency, enhanced extraction performance, and environmental sustainability. The findings of this study not only advance the understanding of DES in phytochemical extraction but also pave the way for broader application of green solvents in the natural products industry. Future research should focus on further optimizing DES formulations and scaling up the extraction process to fully realize the potential of this innovative extraction method in commercial applications. [ABSTRACT FROM AUTHOR]

Introduction: Chlorogenic acid (CGA) has been identified to possess salient antiinflammatory, antioxidant, and anticancer attributes. However, its application is limited by its instability and low bioavailability. Liposomes have been considered effective pharmaceutical delivery vehicles due to their ability to continuously release loaded drugs, improve drug stability, and display good biocompatibility. They can be easily modified by other small molecules to acquire additional biological functions. In this study, we developed and characterized folic acid-TPGS-modified chlorogenic acid liposome (FTCLP) and evaluated its antiinflammatory activity. Methods: The successful encapsulation of CGA within FTCLP was confirmed through examination using electron microscopy, fourier-transform infrared spectroscopy (FTIR), and differential scanning calorimetry (DSC). The in vitro release characteristics of FTCLP were evaluated using the dialysis bag membrane method. Meanwhile, a dextran sulfate sodium (DSS) -induced colitis model was employed to investigate the anti-inflammatory effect of FTCLP and its mechanism. Results: The FTCLP exhibited an encapsulation efficiency (EE) of 84.85 ± 1.20% and a drug loading (DL) of 11.67 ± 0.04%. The particle size of FTCLP was determined to be 150.63 ± 0.71 nm, with a polydispersity index (PDI) of 0.198 ± 0.02 and a zeta potential of 2.61 ± 0.38 mV. The in vitro release profile followed the Higuchi model, indicating sustained-release characteristics. The in vivo study demonstrated that FTCLP treatment was effective in improving the symptoms of DSS-induced inflammatory response, as evidenced by mitigation of weight loss, reduction in the disease activity index (DAI) score, restoration of colon length, and attenuation of colon tissue damage. Furthermore, the levels of pro-inflammatory cytokines, including interferon-gamma (INF-γ), interleukin-1 beta (IL-1β), and interleukin-6 (IL-6), were markedly diminished in both the serum and colon tissue. FTCLP was also observed to suppress the expression of INF-γ, IL-1β, IL-6, tumor necrosis factor-alpha (TNF-α), and nuclear factor kappa B (NF-κB) p65, while concomitantly upregulating the expression of Janus kinase (JAK) and signal transducer and activator of transcription 3 (STAT3). Besides, the administration of FTCLP was found to result in an increase in the abundance of Lactobacillaceae and Peptostreptococcaceae, while decreasing the abundance of Bacteroidaceae, Rikenellaceae, and Helicobacteraceae. Conclusion: Following encapsulation of CGA within liposomes, FTCLP revealed favorable stability and sustained release properties, and enhanced the antiinflammatory effects by modulating multiple inflammation-related biomarkers. FTCLP has the potential to be a safe and effective drug for targeted therapy of colitis. [ABSTRACT FROM AUTHOR]

Idiopathic thrombocytopenic purpura (ITP) is an autoimmune disorder characterized by antiplatelet autoantibodies, thrombocytopenia, and bleeding, however, its treatment options are limited. In this study, a kind of active component, chlorogenic acid compounds (CGAs) from sweetpotato leaves was extracted out to explore its medicinal value and provide novel therapeutic strategies for the treatment of ITP. CGAs was isolated by ionic liquids-ultrasound (IL-UAE), which contains six isomers of chlorogenic acid with total purity of 95.69%. The thrombopoietic effect and mechanism of CGAs were investigated using in silico prediction and experimental validation. The changes of HEL cells morphology in volume and the increase in the total cell percentage of polyploid cells indicated that CGAs could promote megakaryocyte differentiation. Meanwhile, CGAs could promote platelet formation in a murine model of ITP, which was established by injection of antiplatelet antibody. Further quantitative proteomics analysis and Western blot verification revealed that CGAs could activate PI3K/AKT signaling pathway, which confirmed the mechanism prediction. It suggested that CGAs may provide a novel therapeutic strategy that relies on the PI3K/AKT pathway to facilitate megakaryocyte differentiation and platelet production. [ABSTRACT FROM AUTHOR]

DEFENSE NO DEATH 1 (DND1) is a cyclic nucleotide-gated ion channel protein. Earlier, it was shown that the silencing of DND1 in the potato (Solanum tuberosum L.) leads to resistance to late blight, powdery mildew, and gray mold diseases. At the same time, however, it can reduce plant growth and cause leaf necrosis. To obtain knowledge of the molecular events behind the pleiotropic effect of DND1 downregulation in the potato, metabolite and transcriptome analyses were performed on three DND1 silenced lines of the cultivar 'Désirée.' A massive increase in the salicylic acid content of leaves was detected. Concentrations of jasmonic acid and chlorogenic acid and their derivatives were also elevated. Expression of 1866 genes was altered in the same way in all three DND1 silenced lines, including those related to the synthesis of secondary metabolites. The activation of several alleles of leaf rust, late blight, and other disease resistance genes, as well as the induction of pathogenesis-related genes, was detected. WRKY and NAC transcription factor families were upregulated, whereas bHLHs were downregulated, indicating their central role in transcriptome changes. These results suggest that the maintenance of the constitutive defense state leads to the reduced growth of DND1 silenced potato plants. [ABSTRACT FROM AUTHOR]

Introduction: Walnut green husk (WGH) is a waste byproduct from walnut industry. However, it is not well-known about its bioactive effect on human gut health. Methods: This study conducted in vitro digestion and fermentation experiments to study the bioactive effect of WGH. Results: Microbial fermentation was the primary mechanism to efficiently release phenolics and flavonoids, resulting in more excellent antioxidant capacities (DPPH, ABTS, and FRAP assays), which reached a highest value with 14.82 ± 0.01 mg VcE/g DW, 3.47 ± 0.01 mmol TE/g DW, and 0.96 ± 0.07 mmol FeSO4·7H2O/g DW, respectively. The surface microstructure of WGH became loose and fragmented after microbial fermentation. The analytical results of gut microbiota demonstrated that WGH could significantly increase the relative abundance of Proteobacteria in phylum level and Phascolarctobacterium in genus level while certain pro-inflammatory bacteria (such as Clostridium_sensu_stricto_1, Dorea, Alistipes, and Bilophila) was inhibited. Additionally, 1,373 differential metabolites were identified and enriched in 283 KEGG pathways. Of which some metabolites were significantly upregulated including ferulic acid, chlorogenic acid, umbelliferone, scopolin, muricholic acid, and so forth. Discussion: These results indicated that WGH could have antioxidant and antiinflammatory activities in the human gut, which could improve the economical value of WGH in the food industry. [ABSTRACT FROM AUTHOR]

Background: The vulnerability of buffalo sperm to cryoinjury necessitates the improvement of sperm cryo‐resistance as a critical strategy for the widespread use of assisted reproductive technologies in buffalo. Objectives: The main aim of the present study was to evaluate the effects of different concentrations of rutin and chlorogenic acid (CGA) on buffalo semen quality, antioxidant activity and fertility during cryopreservation. Methods: The semen was collected and pooled from the 3 buffaloes using an artificial vagina (18 ejaculations). The pooled sperm were divided into nine different groups: control (Tris‐based extender); 0.4, 0.6, 0.8 and 1 mM rutin (rutin + Tris‐based extender); and 50, 100, 150 and 200 µM CGG (CGA + Tris‐based extender). Sperm kinematics, viability, hypo‐osmotic swelling test, mitochondrial activity, antioxidant activities and malondialdehyde (MDA) concentration of frozen and thawed buffalo sperm were evaluated. In addition, 48 buffalo were finally inseminated, and pregnancy was rectally determined 1 month after insemination. Results: Compared to the control group, adding R‐0.4, R‐0.6, CGA‐100 and CGA‐150 can improve total and progressive motility, motility characteristics, viability, PMF and DNA damage in buffalo sperm. In addition, the results showed that R‐0.4, R‐0.6, CGA‐50, CGA‐100 and CGA‐150 increased total antioxidant capacity, catalase, glutathione peroxidase and glutathione activities and decreased MDA levels compared to the control group. Furthermore, it has been shown that adding 150 µM CGA and 0.6 mM rutin to an extender can increase in vivo fertility compared to the control group. Conclusions: In conclusion, adding rutin and CGA to the extender improves membrane stability and in vivo fertility of buffalo sperm by reducing oxidative stress. [ABSTRACT FROM AUTHOR]

Airborne particulate matter (PM) contains polycyclic aromatic hydrocarbons (PAHs) as primary toxic components, causing oxidative damage and being associated with various inflammatory skin pathologies such as premature aging, atopic dermatitis, and psoriasis. Coffee cherry pulp (CCS) extract, rich in chlorogenic acid, caffeine, and theophylline, has demonstrated strong antioxidant properties. However, its specific anti-inflammatory effects and ability to protect macrophages against PAH-induced inflammation remain unexplored. Thus, this study aimed to evaluate the anti-inflammatory properties of CCS extract on RAW 264.7 macrophage cells exposed to atmospheric PAHs, compared to chlorogenic acid (CGA), caffeine (CAF), and theophylline (THP) standards. The CCS extract was assessed for its impact on the production of nitric oxide (NO) and expression of tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2). Results showed that CCS extract exhibited significant antioxidant activities and effectively inhibited protease and lipoxygenase (LOX) activities. The PAH induced the increase in intracellular reactive oxygen species, NO, TNF-α, IL-6, iNOS, and COX-2, which were markedly suppressed by CCS extract in a dose-dependent manner, comparable to the effects of chlorogenic acid, caffeine, and theophylline. In conclusion, CCS extract inhibits PAH-induced inflammation by reducing pro-inflammatory cytokines and reactive oxygen species (ROS) production in RAW 264.7 cells. This effect is likely due to the synergistic effects of its bioactive compounds. Chlorogenic acid showed strong antioxidant and anti-inflammatory activities, while caffeine and theophylline enhanced anti-inflammatory activity. CCS extract did not irritate the hen's egg chorioallantoic membrane. Therefore, CCS extract shows its potential as a promising cosmeceutical ingredient for safely alleviating inflammatory skin diseases caused by air pollution. [ABSTRACT FROM AUTHOR]

We describe the antioxidant capability of scavenging the superoxide radical of several tea and yerba mate samples using rotating ring–disk electrochemistry (RRDE). We directly measured superoxide concentrations and detected their decrease upon the addition of an antioxidant to the electrochemical cell. We studied two varieties of yerba mate, two varieties of black tea from Bangladesh, a sample of Pu-erh tea from China, and two components, caffeic acid and chlorogenic acid. All of these plant infusions and components showed strong antioxidant activities, virtually annihilating the available superoxide concentration. Using density functional theory (DFT) calculations, we describe a mechanism of superoxide scavenging via caffeic and chlorogenic acids. Superoxide can initially interact at two sites in these acids: the H4 catechol hydrogen (a) or the acidic proton of the acid (b). For (a), caffeic acid needs an additional π–π superoxide radical, which transfers electron density to the ring and forms a HO2− anion. A second caffeic acid proton and HO2− anion forms H2O2. Chlorogenic acid acts differently, as the initial approach of superoxide to the catechol moiety (a) is enough to form the HO2− anion. After an additional acidic proton of chlorogenic acid is given to HO2−, three well-separated compounds arise: (1) a carboxylate moiety, (2) H2O2, and a (3) chlorogenic acid semiquinone. The latter can capture a second superoxide in a π–π manner, which remains trapped due to the aromatic ring, as for caffeic acid. With enough of both acids and superoxide radicals, the final products are equivalent: H2O2 plus a complex of the type [X-acid–η–O2], X = caffeic, chlorogenic. Chlorogenic acid (b) is described by the following reaction: 2 O2•− + 2 chlorogenic acid → 2 chlorogenic carboxylate + O2 + H2O2, and so, it acts as a non-enzymatic superoxide dismutase (SOD) mimic, as shown via the product formation of O2 plus H2O2, which is limited due to chlorogenic acid consumption. Caffeic acid (b) differs from chlorogenic acid, as there is no acidic proton capture via superoxide. In this case, approaching a second superoxide to the H4 polyphenol moiety forms a HO2− anion and, later, an H2O2 molecule upon the transfer of a second caffeic acid proton. [ABSTRACT FROM AUTHOR]

Leaves of large‐leaved lime, Tilia platyphyllos Scop. (Malvaceae), harbor colonies of Eriophyes tiliae (Pagenstecher) (Acari: Eriophyidae), capable of modifying the leaf structure (inducer of nail‐galls) and physiology. The aerial organs of T. platyphyllos have traditionally been consumed for their high antioxidant capacity, related to the rich polyphenol profile. Here, we investigated the polyphenol profiles and antioxidant activity of T. platyphyllos non‐galled leaves and galls to determine the changes induced by the mite in a Chilean environment with high ultraviolet (UV) radiation. Phenolic compound identification in the methanol extracts of non‐galled leaves and galls was carried out through HPLC. Furthermore, the antioxidant activity of the extracts was quantified through spectrophotometry, and it was compared with the standards of the three major phenolic compounds of both organs. Nine phenolic compounds were detected in non‐galled leaves and galls, with no differences between the two organs, except for the absence of gallic acid in galls. The concentration of phenolic compounds did differ significantly between the two conditions. In leaf galls, epicatechin and cyanidin‐3‐glucoside concentrations and antioxidant capacity increased significantly compared to non‐galled leaves. As chlorogenic acid and cyanidin‐3‐glucoside are active phenols responding to UV radiation stress, their concentration in T. platyphyllos leaves could be a response to the high UV radiation occurring in Chile during spring and summer. The phenolic compounds detected here have been reported to be potent antioxidants, that are probably potentiated by E. tiliae for its own protection against UV‐B radiation, as the two most abundant compounds in the galls exhibited the highest capacity to reduce ABTS and DPPH radicals. However, we do not rule out the participation of phenolics in protection against natural enemies of the gall mite, as chlorogenic acid and epicatechin are powerful antifeedants. [ABSTRACT FROM AUTHOR]

As a prevalent neurodegenerative disorder, Parkinson's disease is associated with oxidative stress. Our recent investigations revealed that reactive oxygen species (ROS) and PD-toxins like 6-hydroxydopamine (6-OHDA) can induce neuronal apoptosis through over-activation of Akt signaling. Chlorogenic acid (CGA), a natural acid phenol abundant in the human diet, is well-documented for its ability to mitigate intracellular ROS. In this study, we utilized CGA to treat experimental models of PD both in vitro and in vivo. Our study results demonstrated that SH-SY5Y and primary neurons exhibited cell apoptosis in response to 6-OHDA. Pretreatment with CGA significantly attenuated PD toxins-induced large amount of ROS, inhibiting Erk1/2 activation, preventing Akt inhibition, and hindering neuronal cell death. Combining the Erk1/2 inhibitor U0126 with CGA could reverse 6-OHDA-induced Akt inhibition, ROS, and apoptosis in the cells. Crucially, the Akt activator SC79 and ROS scavenger NAC both could eliminate excessive ROS via Akt and Erk1/2 signaling pathways, and CGA further potentiated these effects in PD models. Behavioral experiments revealed that CGA could alleviate gait abnormalities in PD model mice. The neuroprotective effects have been demonstrated in several endocrine regions and in the substantia nigra tissue, which shows the positive tyrosine hydroxylase (TH). Overall, our results suggest that CGA prevents the activation of Erk1/2 and inactivation of Akt by removing excess ROS in PD models. These findings propose a potential strategy for mitigating neuronal degeneration in Parkinson's disease by modulating the Akt/Erk1/2 signaling pathway through the administration of CGA and/or the use of antioxidants to alleviate oxidative stress. • CGA effectively mitigates the axonal damage induced by 6-OHDA in neuronal cells. • CGA inhibits excessive ROS production via the AKT/Erk1/2 signal pathway, preventing 6-OHDA-induced neuronal apoptosis. • CGA ameliorates dyskinesia of the A53T mice. • CGA mitigates the neuronal apoptosis and the histopathologic aberrations via the AKT/Erk1/2 signal pathway in the A53T mice. [ABSTRACT FROM AUTHOR]

Cornus mas L. (Cornelian cherry, CM) fruits were dehydrated by solar-drying (SD) and freeze-drying (FD), and in addition to sugar and mineral contents, the free and insoluble-bound phenolics were determined in fresh and dried fruits. After subjecting the sample to simulated in vitro digestion, the change of free and bound phenolics at gastric and intestinal digestion steps was evaluated in fresh and dried CM fruits. In fresh CM fruits, the total phenolic content (TPC) was dominated by the bound fraction, whereas the contribution of free phenolics to the total content (free + bound) became more dominant (731–1439 mg GAE/100 g dw) in the dried fruits. The bioaccessibility (BI%) of TPC from fresh CM after digestion was 193%, whereas it was 18.60 and 48.02% for SD and FD fruits, respectively. The contribution free fraction to the total TPC value was around 28% in nondigested fresh samples and increased to 94% in digested samples; however, in dried samples, it was 64% prior to digestion and only increased to 70% in digested samples. A total of 17 phenolic compounds were identified in CM fruits: chlorogenic acid, caffeic acid, epicatechin, quercetin, cyanidin-3-O-glucoside, and pelargonidin 3-O-glucoside were only detected in the free fraction; gallic acid, vanillic acid, ferulic acid, and kaempferol were detected in higher amounts in the bound fraction. The quantity of detected phenolics in the nondigested sample generally decreased from the gastric to the intestinal stage of digestion. The release of phenolics from the fruit matrix and their degradation occurred simultaneously during digestion, and this could be affected by the state of the fruit, e.g., fresh or dried. [ABSTRACT FROM AUTHOR]

Coffee is one of the most popular beverages in the world, and at present, specialty coffees are developing for better tastes, flavors, or functions. Fermented coffees also reflect this trend and some brands are commercialized. Unlike general fermented coffees, we tried to ferment coffee extract with several lactic acid bacteria. Finally, we selected three strains that persisted in the coffee extract, and show strong antimicrobial activity, for the fermentation starters. The strains were identified as Pediococcus pentosaceus (KNUT 0384), Lacticaseibacillus paracasei (CJNU 1840), and Lactiplantibacillus plantarum (CJNU 0441) based on 16S rRNA gene sequences. During the fermentation process, the total acidities (p < 0.05 vs. the control, non-fermented coffee extract) and cell masses increased, which indicates that the bacteria metabolized properly in the extract. Furthermore, the fermented coffee extracts showed increased antimicrobial activities against Listeria monocytogenes (p < 0.05 vs. the control) and Streptococcus mutans (p < 0.05 vs. the control), increased anti-oxidative activities (p < 0.05 vs. the control, except for the KNUT 0384 sample), decreased caffeine content (p < 0.05 vs. the control, except for the KNUT 0384 sample), and increased chlorogenic acid content (p < 0.05 vs. the control). Taken together, the fermented coffee extracts with the selected lactic acid bacteria could be specialty coffees where several functions are improved when compared with a control coffee extract. [ABSTRACT FROM AUTHOR]

This study analyzes the effects on body composition and variables related to metabolic syndrome of two coffees with different degree of roasting and phenolic content. Sixty participants with body mass index between 25 and 35 kg/m2 and a median age of 51.0 years (Interquartile range 46.3–56) were recruited. The study was a controlled, randomized, single-blind crossover trial consisting in drinking three cups/day of roasted coffee (RC) or lightly roasted coffee (LRC) during 12 weeks with 2-week wash-out stages before each coffee intervention. LRC contained ≈400 mg of hydroxycinnamic acids and ≈130 mg of caffeine per 200 mL/cup while RC contained ≈150 mg of hydroxycinnamic acids and ≈70 mg of caffeine per 200 mL/cup. Along the study, in each of the six visits, blood pressure, body composition by bioimpedance, anthropometric measurements, and blood biochemistry were analyzed. The mean differences and p values were calculated using a linear mixed model (JASP.v.0.18.0.3). A total of 38 participants completed the study. After the consumption of both coffees, fat mass and body fat percentage (LRC: −1.4%, p < 0.001; RC: −1.0%, p = 0.005) were reduced, whereas muscle mass and muscle mass percentage slightly increased (LRC: 0.8%, p < 0.001; RC: 0.7%, p = 0.002). The decrease in fat percentage was greater with LRC compared to RC (−0.8%; p = 0.029). There were no significant changes in metabolic syndrome variables or in body weight. In conclusion, LRC was slightly superior at inducing changes in body composition. [ABSTRACT FROM AUTHOR]

Due to containing more than 100 g/L of sugar, liqueurs are high-calorie spirits, providing consumers only 'empty calories' with no nutritional value or benefit. Elderflower is intriguing as a possible ingredient in creative alcoholic beverages, yet it is rarely used for the manufacturing of commercial spirits. This is the first study aiming to use steviol glycosides powder (SGP) to reduce the amount of sugar in elderflower liqueur. Optimal sweetness of liqueur and equivalent sweetness of SGP were determined by sensory evaluation. Four samples, one prepared with only sucrose as a sweetener and three with 25%, 50%, and 75% of sucrose replaced with commercial SGP, were subjected to consumer acceptance tests. The overall acceptance of all samples was near and in the range of liking, with hedonic scores ranging from 5.6 to 6.6, indicating the promising liqueurs' commercial potential. Superior concentrations of chlorogenic acid and caffeic acid were found in liqueurs. The total phenolic content of liqueurs increased as SGP concentration increased, while the sample with the highest SGP content had the greatest capacity to scavenge DPPH. Functional and sensory properties of novel liqueurs indicate the promising potential of sugar substitution by SGP in liqueur production. [ABSTRACT FROM AUTHOR]

This research focused on the impact of drying techniques on the physicochemical and aromatic profile of honey coffee, which involves simultaneous fermentation and drying of coffee beans. Sun drying (SN), low temperature and low humidity drying (LTLH), polyhouse/solar tunnel drying (SD), and shade drying (SH) were carried out. Insignificant effect on physical quality was recorded. Chemical characterization of the raw coffee revealed that the LTLH-dried beans had higher carbohydrates (15.53%), which affected the coffee aroma. The roasted coffee contained caffeine (2.86%) and chlorogenic acid (2.50%), the primary bioactive components, contributing to strength, astringency and bitterness. The coffee brews had improved chemical metabolites such as carbohydrates (16.06%), total polyphenols (6.36%), flavonoids (0.7%) and antioxidants (5.4%). Furthermore, PCA correlates a total variance of 99.2% on green coffee, 97.3% on roasted coffee and 99.8% on brew. SPME–GC–MS revealed 75 volatiles in the roasted coffee and 95 in the infusions. Individual value plot graphs were introduced to assess the effect of volatiles on drying modes. Pyrazines and ketones were key molecules contributing to roasted walnut notes with fruity, floral and honey undertones. E-Nose profiles excellently overlaid GCMS results. SIMCA represented a positive correlation between LTLH coffee and volatile compounds. Thus, the LTLH technique (45–50 °C and 30–40% RH) for honey coffee facilitated an appropriate fermentation of coffee beans thereby improving the overall quality which, in turn, leads to global attention for innovative coffee processing technologies. [ABSTRACT FROM AUTHOR]

Exposure to fine particulate matter with an aerodynamic diameter of less than 2.5 μm (PM2.5) can cause oxidative damage and apoptosis in the human skin. Chlorogenic acid (CGA) is a bioactive polyphenolic compound with antioxidant, antifungal, and antiviral properties. The objective of this study was to identify the ameliorating impact of CGA that might protect human HaCaT cells against PM2.5. CGA significantly scavenged the reactive oxygen species (ROS) generated by PM2.5, attenuated oxidative cellular/organelle damage, mitochondrial membrane depolarization, and suppressed cytochrome c release into the cytosol. The application of CGA led to a reduction in the expression levels of Bcl‐2‐associated X protein, caspase‐9, and caspase‐3, while simultaneously increasing the expression of B‐cell lymphoma 2. In addition, CGA was able to reverse the decrease in cell viability caused by PM2.5 via the inhibition of extracellular signal‐regulated kinase (ERK). This effect was further confirmed by the use of the mitogen‐activated protein kinase kinase inhibitor, which acted upstream of ERK. In conclusion, CGA protected keratinocytes from mitochondrial damage and apoptosis via ameliorating PM2.5‐induced oxidative stress and ERK activation. [ABSTRACT FROM AUTHOR]

As a traditional Chinese medicine (TCM), Cortex Periplocae (CP) has a wide range of pharmacological effects, as well as toxic side effects. The main toxic components of it are cardiac glycosides, which tend to cause cardiotoxicity. Currently, it has also been reported in studies to cause hepatotoxicity, but it is not clear whether the hepatotoxicity is related to the toxicity caused by the reactive metabolites. This study aims to investigate the target components of CP that generate reactive metabolic toxicity. The fluorescent probe method was used to detect glutathione (GSH)-trapped reactive metabolites in a co-incubation system of CP extract with rat liver microsomes. Identification of GSH conjugates was performed by LC–MS/MS and that of the possible precursor components that produce reactive metabolites was conducted by UPLC–Q-TOF/MS. Cell viability assays were performed on HepG2 and L02 cells to determine the cytotoxicity of the target components. The findings of our study demonstrate that the extract derived from CP has the ability to generate metabolites that exhaust the intracellular GSH levels, resulting in the formation of GSH conjugates and subsequent cytotoxic effects. Through the utilization of the UPLC–Q-TOF/MS technique, we were able to accurately determine the molecular weight of the precursor compound in CP to be 355.1023. The primary evidence to determining the GSH conjugetes relies on the appearance of characteristic product ions resulting from central neutral loss (CNL) scanning of 129 Da and product scanning of m/z 660 in the positive MS/MS spectrum. Through analysis, it was ultimately ascertained that the presence of chlorogenic acid (CGA) and its isomers, namely neochlorogenic acid (NCGA) and cryptochlorogenic acid (CCGA), could lead to the production of GSH conjugates, resulting in cytotoxicity at elevated levels. Taking these findings into consideration, the underlying cause for the potential hepatotoxicity of CP was initially determined. [ABSTRACT FROM AUTHOR]

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Recently, increased attention has been paid to the raw materials of plants as a source of biologically active substances. Black rosehip (Rosa pimpinellifolia L.) fruits could be a good resource for potential functional components in the food, cosmetic and pharmaceutical industries. Also, drying can influence the composition of heat-sensitive phytochemicals. However, less attention is given to comparing black rosehip bioactive compounds particularly compositions of fatty acid, amino acids, and phenolic content in fresh and dried fruits. So in this study, the amino acid constituents (by amino acid analyzer), fatty acids (by GC–MS), mineral elements (by atomic absorption spectrometer), antioxidant (by DPPH) and phenolic compounds (by HPLC) present in fresh and dried fruits of black rosehip naturally grown in Iran were comprehensively investigated. The results showed that dried fruits had a lower level moisture by 51.55%, and a higher level of total phenolic compounds and total sugar by 786.20 mg GAE/100 g and 15.77 g/100 g, respectively. Chlorogenic acid and gallic acid were the major phenolic compounds (109.3 mg/g). Whilst, linoleic acid, oleic acid, and arachidic acid (85.34%) were the most dominant fatty acids. The most dominant amino acids were glutamic acid, phenylalanine, and arginine (29.41 g/100 g). Also, Fe and Mn as micro-elements were the most dominant elements. In general, the results illustrated the potentials and differences of black rosehip fruits grown in the Arasbaran region as promising resources for food sources, pharmaceutics, cosmetics, and breeding programs. Also, these findings confirm that black rosehip fruits contain significant amounts of secondary metabolites that may positively affect human health. [ABSTRACT FROM AUTHOR]

AbstractCousinia birandiana is one of the widely spread species of the Asteraceae family. In this study, the total phenolic and flavonoid contents, the inhibitory effect on α-amylase, α-glucosidase, acetylcholinesterase and butyrylcholinesterase enzymes, the antioxidant capacity (by DPPH, ABTS, FRAP, CUPRAC tests) and the phytochemical content (by LC-HRMS) of the ethanol extract and fractions were studied. As a result, it was shown that ethanol extract has better enzyme inhibitory effects on α-amylase, AChE and BChE enzymes with 300.000, 380.605 and 133.567 µg/mL IC50 values. Besides that, the highest phenolic and flavonoid content was found in the ethyl acetate fraction (735.986 mgGAE/gextract and 952.317 mgCA/gextract, respectively). It is observed that the same fraction has better antioxidant activity in DPPH (IC50 =112.701 mg/mL), ABTS (9.494 µM Trolox/gextract) and CUPRAC (1120.454 mgGAE/gextract) tests and the n-hexane fraction has a higher FRAP (1351.199 mmol Fe2+/gextract) antioxidant effect. According to the phytochemical analyses quinic acid, chlorogenic acid and isoquercitrin were found as major compounds in the ethanol extract and fractions. In addition, the same assays were carried out with these major substances. So, this plant can be viewed as a promising bio-resource for pharmaceuticals and biomedical products. [ABSTRACT FROM AUTHOR]

Artichokes (Cynara scolymus L.) are valuable foods, thanks to their health benefits, but they generate significant waste during their production, harvesting, and processing, which poses sustainability issues. This study applied an agroprospecting approach to convert Tema artichoke biowaste (TB) into valuable resources, starting from a global perspective of the production chain to the targeted applications based on chemical and biological analysis. The major TB was identified in the outer bracts of the immature flower heads, which were collected throughout the harvesting season, extracted, and analyzed. The most abundant compounds were phenolic acids including chlorogenic acid and caffeoylquinic derivatives. Among flavonoids, cynaroside was the most abundant compound. Multivariate analysis distinguished batches by collection period, explaining 77.7% of the variance, with most compounds increasing in concentration later in the harvest season. Subsequently, TB extracts were analyzed for their potential in wound healing and anti-aging properties. Fibroblasts were used to assess the effect of selected extracts on cell migration through a scratch wound assay and on cellular senescence induced by etoposide. The results show a significant decrease in senescence-associated β-galactosidase activity, γH2AX nuclear accumulation, and both p53 and p21 protein levels. Overall, this study ascribes relevant anti-skin aging effects to TB, thus increasing its industrial value in cosmeceutical and nutraceutical applications. [ABSTRACT FROM AUTHOR]

Hypericum perforatum transformed shoot lines (TSL) regenerated from corresponding hairy roots and non-transformed shoots (NTS) were comparatively evaluated for their phenolic compound contents and in vitro inhibitory capacity against target enzymes (monoamine oxidase-A, cholinesterases, tyrosinase, α-amylase, α-glucosidase, lipase, and cholesterol esterase). Molecular docking was conducted to assess the contribution of dominant phenolic compounds to the enzyme-inhibitory properties of TSL samples. The TSL extracts represent a rich source of chlorogenic acid, epicatechin and procyanidins, quercetin aglycone and glycosides, anthocyanins, naphthodianthrones, acyl-phloroglucinols, and xanthones. Concerning in vitro bioactivity assays, TSL displayed significantly higher acetylcholinesterase, tyrosinase, α-amylase, pancreatic lipase, and cholesterol esterase inhibitory properties compared to NTS, implying their neuroprotective, antidiabetic, and antiobesity potential. The docking data revealed that pseudohypericin, hyperforin, cadensin G, epicatechin, and chlorogenic acid are superior inhibitors of selected enzymes, exhibiting the lowest binding energy of ligand–receptor complexes. Present data indicate that H. perforatum transformed shoots might be recognized as an excellent biotechnological system for producing phenolic compounds with multiple health benefits. [ABSTRACT FROM AUTHOR]

Natural products received much attention as an environmentally beneficial solution for pest management. Therefore, the extracts of invasive silverleaf nightshade (Solanum elaeagnifolium Cav.) weeds using their berries parts (seeds, peels and mucilage) supported by bioassay-guided fractionation were tested against both the greater wax moth (Galleria mellonella) and Erwinia carotovora pv. carotovora causes of the blackleg of potatoes. The seeds and peels of S. elaeagnifolium were successively extracted by maceration using dichloromethane (DCM), ethyl acetate (EtOAc), and ethanol (EtOH), respectively. While, its mucilage was extracted using EtOAc. The successive EtOH extract of the plant seeds had promising inhibition efficacy and the best minimal inhibition concentration (MIC) of 50 µg/ml against E. Carotovora amongst other extracts (DCM and EtOAc of the plant berries parts). Depending on dose response activity, EtOH extract had G. mellonella larval mortality and pupal duration rates (LC50; 198.30 and LC95; 1294.73 µg/ml), respectively. Additionally, this EtOH extract of seeds was fractionated using preparative TLC to three characteristic bands. The insecticidal and bacterial activities of these isolated bands (SEA, SEB, and SEC) were evaluated at a dose of 100 µg/ml, causing mortality by 48.48, 62.63 and 92.93% (G. mellonella larvae) and inhibition by 15.22, 0.00 and 31.66 mm (E. carotovora), respectively. Moreover, the separated major three bands were tentatively identified using LC-ESI-MS analysis revealing the presence of two phenolic acids; chlorogenic acid (SEA) and dicaffeoyl quinic acid (SEB) in addition to one steroidal saponin (SEC) annotated as borassoside E or yamoscin. Finally, the plant seeds' successive EtOH extract as well as its active constituents, exhibited potential broad-spectrum activity and the ability to participate in future pest management initiatives. A field study is also recommended to validate its bio-efficacy against selected pests and to develop its formulations. [ABSTRACT FROM AUTHOR]

SUMMARY UV‐B radiation can induce the accumulation of many secondary metabolites, including flavonoids, in plants to protect them from oxidative damage. BRI1‐EMS‐SUPPRESSOR1 (BES1) has been shown to mediate the biosynthesis of flavonoids in response to UV‐B. However, the detailed mechanism by which it acts still needs to be further elucidated. Here, we revealed that UV‐B significantly inhibited the transcription of multiple transcription factor genes in tobacco, including NtMYB27, which was subsequently shown to be a repressor of flavonoids synthesis in tobacco. We further demonstrated that NtBES1 directly binds to the E‐box motifs present in the promoter of NtMYB27 to mediate its transcriptional repression upon UV‐B exposure. The UV‐B‐repressed NtMYB27 could bind to the ACCT‐containing element (ACE) in the promoters of Nt4CL and NtCHS and served as a modulator that promoted the biosynthesis of lignin and chlorogenic acid (CGA) but inhibited the accumulation of flavonoids in tobacco. The expression of NtMYB27 was also significantly repressed by heat stress, suggesting its putative roles in regulating heat‐induced flavonoids accumulation. Taken together, our results revealed the role of NtBES1 and NtMYB27 in regulating the synthesis of flavonoids during the plant response to UV‐B radiation in tobacco. [ABSTRACT FROM AUTHOR]

Sweet potatoes are extremely vulnerable to mechanical wounds during harvesting and postharvest handling. It is highly necessary to take measures to accelerate wound healing. The effect of 20 g L−1 of ascorbic acid (AA) treatment on the wound healing of sweet potatoes and its mechanisms were studied. The results validated that AA treatment significantly reduced the weight loss rate and disease index. AA treatment effectively enhanced the formation speed of lignin and SPP at the wound sites, decreased the MDA content, and maintained the cell membrane integrity. AA enhanced the activities of PAL, C4H, 4CL, CAD, and POD and increased the contents of chlorogenic acid, caffeic acid, sinapic acid, ferulic acid, cinnamic acid, p-coumaryl alcohol, sinapyl alcohol, coniferyl alcohol, and lignin. Based on a transcriptomic analysis, a total of 1200 genes were differentially expressed at the sweet potato wound sites by the AA treatment, among which 700 genes were upregulated and 500 genes were downregulated. The KEGG pathway analysis showed that the differentially expressed genes were mainly involved in phenylalanine, tyrosine, and tryptophan biosynthesis; phenylpropanoid biosynthesis; and other wound healing-related pathways. As verified by a qRT-PCR, the AA treatment significantly upregulated the gene expression levels of IbSKDH, IbADT/PDT, IbPAL, and Ib4CL at the wound sties. [ABSTRACT FROM AUTHOR]

Plant-growth-promoting bacteria (PGPB) have beneficial effects on plants. They can promote growth and enhance plant defense against abiotic stress and disease, and these effects are associated with changes in the plant metabolite profile. The research problem addressed in this study was the impact of inoculation with PGPB on the metabolite profile of Salicornia europaea L. across controlled and field conditions. Salicornia europaea seeds, inoculated with Brevibacterium casei EB3 and Pseudomonas oryzihabitans RL18, were grown in controlled laboratory experiments and in a natural field setting. The metabolite composition of the aboveground tissues was analyzed using GC–MS and UHPLC–MS. PGPB inoculation promoted a reconfiguration in plant metabolism in both environments. Under controlled laboratory conditions, inoculation contributed to increased biomass production and the reinforcement of immune responses by significantly increasing the levels of unsaturated fatty acids, sugars, citric acid, acetic acid, chlorogenic acids, and quercetin. In field conditions, the inoculated plants exhibited a distinct phytochemical profile, with increased glucose, fructose, and phenolic compounds, especially hydroxybenzoic acid, quercetin, and apigenin, alongside decreased unsaturated fatty acids, suggesting higher stress levels. The metabolic response shifted from growth enhancement to stress resistance in the latter context. As a common pattern to both laboratory and field conditions, biopriming induced metabolic reprogramming towards the expression of apigenin, quercetin, formononetin, caffeic acid, and caffeoylquinic acid, metabolites that enhance the plant's tolerance to abiotic and biotic stress. This study unveils the intricate metabolic adaptations of Salicornia europaea under controlled and field conditions, highlighting PGPB's potential to redesign the metabolite profile of the plant. Elevated-stress-related metabolites may fortify plant defense mechanisms, laying the groundwork for stress-resistant crop development through PGPB-based inoculants, especially in saline agriculture. [ABSTRACT FROM AUTHOR]

Gelsemium sempervirens (L.) J.St.-Hil. is an evergreen shrub occurring naturally in North and Middle America. So far, more than 120 alkaloids have been identified in this plant in addition to steroids, coumarins and iridoids, and its use in traditional medicine has been traced back to these compound classes. However, a comprehensive phytochemical investigation of the plant with a special focus on further compound classes has not yet been performed. Therefore, the present study aimed at an extensive HPLC-MSn characterization of secondary metabolites and, for the first time, reports the occurrence of various depsides and phenolic glycerides in G. sempervirens roots and rhizomes, consisting of benzoic and cinnamic acid derivatives as well as dicarboxylic acids. Furthermore, mono- and disaccharides were assigned by GC-MS. Applying the Folin–Ciocalteu assay, the phenolic content of extracts obtained with different solvents was estimated to range from 30 to 50% calculated as chlorogenic acid equivalents per g dry weight and was related to the DPPH radical scavenging activity of the respective extracts. Upon lactic acid fermentation of aqueous G. sempervirens extracts, degradation of phenolic esters was observed going along with the formation of low-molecular volatile metabolites. [ABSTRACT FROM AUTHOR]

Ilex paraguariensis, commonly known as yerba mate, is a plant belonging to the holly genus Ilex and the Aquifoliaceae family, indigenous to South America, and is used for the production of yerba mate. Yerba mate is renowned for its abundance of essential nutrients and bioactive compounds. Based on test results, it can be assumed that the selection of raw material for the preparation of extracts as well as the extraction method significantly influence the final content of biologically active compounds in the extracts. Consequently, this variability impacts the ultimate concentration of biologically active substances within the end product, potentially influencing human consumption. The present study aimed to quantify and compare the content of selected biological active compounds in supplements and products containing I. paraguariensis extracts, along with organic yerba mate dried through a smoke-free process, available in the European market (P-1–P-10). The evaluation focused on antioxidant substances such as neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, 4-feruloylquinic acid, isochlorogenic acid, rutoside astragalin, and caffeine. Additionally, the concentration of specific macro and trace elements was ascertained. The antioxidant compound makeup differs between methanol-extracted samples and aqueous extracts. In both cases, methanol extracts, particularly those in instant and traditional herb forms, showed the highest content of organic compounds with antioxidant properties (such as phenolic compounds and caffeine). The highest content of chlorogenic acid was detected in both methanol (14.7412 mg/g d.w.) and water (8.3120 mg/g d.w.) extracts in product P-4. The caffeic acid content ranged from 0.1491 mg/g d.w. to 1.7938 mg/g d.w. in methanol extracts and from 0.0760 mg/g d.w. to 0.4892 mg/g d.w. in water extracts. The neochlorogenic acid content ranged from 2.6869 to 23.9750 mg/g d.w. in ethanol extracts and from 0.4529 to 10.2299 mg/g d.w. in water extracts. Therefore, the traditional preparation of yerba mate as a water infusion does not fully exploit the raw material's potential. Among the tested products, only the dietary supplement in capsule form contained protocatechuic acid, which was not present in any other tested products. Conversely, compounds characteristic of yerba mate found in other preparations were absent in this supplement. The caffeine content was also the lowest in this product. The determined content of active substances did not consistently match the declarations made by producers if stated on the packaging. [ABSTRACT FROM AUTHOR]

Spent Coffee Ground (SCG) is the main coffee industry by-product, rich in dietary fibers and polyphenols. The extractable material of SCG was fractionated, and the phenolic compounds were identified and quantified. Chlorogenic and neochlorogenic acids were identified as the main phenolic components, and the Total Phenolic Content (TPC) of SCG was determined to be 2.16% (dry SCG basis). Furthermore, SCG was characterized in terms of Total Dietary Fiber content, which amounted to 66%. The SCG was valorized for the development of a bakery product (cookie) enhanced with fiber and bioactive polyphenols. Cookies were produced with the addition of 4% and 7% dry and defatted SCG (baked cookie basis). The produced cookie prototypes presented TPC and dietary fiber dependent on the addition level of SCG. TPC values were determined at 588 and 1017 ppm, while dietary fiber values were at 2.7 and 4.6%, respectively. The shelf life of the cookies was monitored over 143 days at three different temperatures (25 °C, 35 °C, and 45 °C) in terms of texture (hardness), color, Peroxide Value (PV), and TPC. It was observed that the PV value significantly increased in samples with incorporated SCG, stored at 45 °C, while in those stored at 25 °C and 35 °C, PV remained at low levels. The TPC of the SCG-enriched samples remained practically constant during the shelf life analysis, while color and hardness increased (mathematically modeled). SCG-added cookies were characterized by increased darkness, increased hardness, and a mild (desirable) coffee flavor. The overall sensory impression scores for 0%, 4%, and 7% SCG-added cookies were 7.5, 8.0, and 8.2, respectively. Based on sensory evaluation test results, the shelf lives of 0%, 4%, and 7% SCG at 25 °C were 359, 435, and 471 days, respectively. Overall, SCG is a potentially valuable ingredient that can be used to develop innovative food (baked) products with enhanced nutritional value and increased shelf life. [ABSTRACT FROM AUTHOR]

Stable ternary complexes of lactoferrin (LF), high methoxyl pectin (HMP) and chlorogenic acid (CGA) were synthesised and characterised. Soluble LF–HMP complexes were prepared at pH 5, 6 and 7 using established phase diagram. CGA bound to ternary complexes at moderate binding strength (Ka: 103–105 L/M). LF, HMP and CGA interacted via noncovalent interactions like hydrophobic interaction. Ternary complexes showed particle size of 135–462 nm (except for 0.05% CGA at pH 5) and zeta‐potential from −17 to −22 mV. Ternary complexes had superior antioxidant capacity over binary ones (P < 0.05). The ternary complexes may find application in functional beverages. [ABSTRACT FROM AUTHOR]

Aim: The aim of the present study is to show how sodium nitrite alters the histology of submandibular salivary glands and livers of Albino rats, as well as how chlorogenic acid may have therapeutic benefits. Methods: A sample size of thirty male Sprague Dawley Albino rats weighing between 100 and 150 g (5-6 weeks old) was randomly allocated into 3 equal groups. Group I: rats were used as controls and were given phosphate buffer solution, whereas Group II: rats were given an 80 mg/kg sodium nitrites (SN) daily dissolved in distilled water. The rats in Group III were given a daily dose of 80 mg/kg SN dissolved in distilled water and after 6 hours each rat received 50 mg/mL freshly prepared chlorogenic acid (CGA) every other day. For 12 weeks, all treatment modalities will be administered orally, every day. After the experiment, all rats were euthanized. Samples from salivary glands and livers were processed and stained with H&E and interleukin 6 (IL 6). Malondialdehyde (MDA) and superoxide dismutase (SOD) enzymes were detected using an ELISA assay. Results: Groups III had nearly comparable findings to Group I regarding histological pattern with normal submandibular glands and livers features. Group III salivary gland treated with CGA exhibited higher SOD levels (20.60±4.81 U/g) in comparison to the SN group, and lower MDA levels (111.58±28.28 nmol/mg) in comparison to the SN treated samples. In comparison to the SN group, CGA treatment significantly reduced MDA levels in liver samples (167.56±21.17 nmol/mg) and raised SOD (30.85±6.77 U/g). Conclusions: Chlorogenic acid has a protective effect against salivary gland and liver toxicity induced by SN in rats. This was mediated via the anti-inflammatory and antioxidative properties of CGA and the restoration of oxidant/antioxidant balance in rat salivary gland and liver. [ABSTRACT FROM AUTHOR]

This study explored the effects of chlorogenic acid (CGA) on antioxidant capacity, immunity, and intestinal health of spotted sea bass Lateolabrax maculatus. Spotted sea bass were fed six diets: a normal-fat diet (NFD) with 11.9% lipid; HFD with 17% lipid (HFD); and HFD supplemented with 100, 200, 300, and 400 mg/kg CGA (HFD1, HFD2, HFD3, and HFD4 respectively) for 7 weeks. Results showed that HFD-feeding significantly decreased CAT and T-SOD activity compared to the containing-CGA groups. Dietary supplementation with 300 mg/kg CGA significantly reduced MDA content and increased T-AOC compared to the HFD group. HFD supplemented with 100 and 200 mg/kg CGA significantly enhanced GSH activity compared to the HFD group. The activity of ACP and AKP significantly increased with 200 mg/kg CGA treatment compared to the HFD group. Compared to the HFD group, dietary CGA significantly increased the levels of LZM. CGA supplementation at 100, 200, and 300 mg/kg CGA significantly increased the levels of complement C3 compared to that in the HFD group. Supplementation of 300 and 400 mg/kg CGA significantly improved the lipase activity in the foregut compared with the HFD group. For histological analysis, HFD-feeding led to a reduction in muscular thickness, villi length, and density of the midgut compared to the NFD group, while the addition of CGA can improve the negative effects induced by HFD. Additionally, HFD-feeding increased both the diversity and abundance of intestinal bacteria in the midgut compared to the NFD diet, while contrasting results were observed in the hindgut. Furthermore, the addition of CGA mitigated the reduction in hindgut bacterial diversity induced by HFD. Overall, the present study indicated that dietary CGA plays a positive role in antioxidant capacity, immunity, and intestinal health of fed HFD. [ABSTRACT FROM AUTHOR]

Corn starch-chlorogenic acid complexes were prepared under 60 °C, 70 °C, 80 °C and 90 °C, respectively. The chlorogenic acid content, complex index ( CI), crystal structure, in-vitro digestibility were studied, to investigate the effect of temperature on the formation and digestibility of corn starch-chlorogenic acid complexes during hydrothermal treatment. The results showed that as the heat temperature increased, the CI of starch increased from 4.27% to 10. 65% gradually, the chlorogenic acid content in the complexes was increased first and then decreased with a highest content of 6.13 mg/ g being prepared at 80 °C. While excessive chlorogenic acid could promote the formation of double helix structure in corn starch, it hindered the reformation of ordered crystal structures. The structural changes in corn starch-chlorogenic acid complexes as the heat temperature increased would gradually enhance their gelatinization stability and resistance to digestion, and the resistant starch (RS) content in the complexes increased from 20.49% to 30.69%. Therefore, the interaction between corn starch and chlorogenic acid could be adjusted by controlling the heat temperature, which in turn modulates the digestibility of corn starch. [ABSTRACT FROM AUTHOR]

Moringa oleifera (M. oleifera) is a plant widely used for its beneficial properties both in medical and non-medical fields. Because they produce bioactive metabolites, plants are a major resource for drug discovery. In this study, two different cultivars of leaves of M. oleifera (Salento and Barletta) were obtained by maceration or microwave-assisted extraction (MAE). We demonstrated that extracts obtained by MAE exhibited a lower cytotoxic profile compared to those obtained by maceration at concentrations ranged from 25 to 400 µg/mL, on both Vero CCL-81 and Vero/SLAM cells. We examined their antiviral properties against two viruses, i.e., the human coronavirus 229E (HCoV-229E) and measles virus (MeV), which are both responsible for respiratory infections. The extracts were able to inhibit the infection of both viruses and strongly prevented their attack and entry into the cells in a range of concentrations from 50 to 12 µg/mL. Particularly active was the variety of Salento that registered a 50% inhibitory concentration (IC50) at 21 µg/mL for HCoV-229E and at 6 µg/mL for MeV. We identified the presence of several compounds through high performance liquid chromatography (HPLC); in particular, chlorogenic and neochlorogenic acids, quercetin 3-O-β-d-glucopyranoside (QGP), and glucomoringin (GM) were mainly observed. In the end, M. oleifera can be considered a promising candidate for combating viral infections with a very strong action in the early stages of viral life cycle, probably by destructuring the viral particles blocking the virus–cell fusion. [ABSTRACT FROM AUTHOR]

• Successfully established the callus induction system of Saxifraga stolonifera Meerb. • The best sterilization method of explants and the ratio of plant growth regulators for callus induction were selected. • SA and SNP could increase the content of secondary metabolites in callus. • This study has certain reference significance for the commercial production of Saxifraga stolonifera Meerb. Elicitation is a well-known strategy for increasing the accumulation of secondary metabolites in callus cultures. In this study, a callus induction system for Saxifraga stolonifera Meerb. was developed, and elicitors were used to enhance secondary metabolite production. Tender leaves of S. stolonifera were used as explants for in vitro culture. We examined callus induction characteristics of S. stolonifera and its response to various elicitors by assessing the sterilization conditions of the explants, varying hormone ratios, and determining the optimal concentrations of salicylic acid (SA), Sodium nitroprusside (SNP) and methyl jasmonate (MeJA). The results revealed that sterilizing the explants with 70% ethanol for 40 s, followed by 0.1% mercury chloride for 10 min, resulted in a survival rate of 82.22% during tissue culture. Following these sterilization methods, callus formation was successfully induced on Murashige and Skoog medium supplemented with 2 mg/L 6-benzylaminopurine and 0.15 mg/L naphthaleneacetic acid, achieving an induction rate of 88.88%. Additionally, the study identified SA as the most effective elicitor for increasing the levels of gallic acid, protocatechuic acid, and quercitrin in the callus, with optimal concentrations of salicylic acid being 60, 40, and 60 μM, respectively. SNP at 40 μM, 80 μM, and 20 μM concentrations was identified as the best elicitor for promoting the accumulation of bergenin, chlorogenic acid, and quercetin, respectively. This study established appropriate conditions for cultivating S. stolonifera callus and identified the optimal elicitor concentrations for maximizing secondary metabolite production. [ABSTRACT FROM AUTHOR]

As storage time increases, the quality of traditional Chinese medicines (TCMs) may change, and stability is an essential aspect of ensuring the safety and efficacy of TCMs. In this study, the effects of different storage times on the stability of 12 decoction pieces were evaluated. High-performance liquid chromatography was used to determine the contents of the active components in the 12 decoction pieces. The chemical composition data were analyzed using fingerprinting and clustering heatmap (CH). Results showed that during storage, significant variations (relative standard deviation > 10%) were observed in the levels of paeoniflorin in Paeoniae Radix Alba and Paeoniae Radix Rubra, hesperidin in Citri Reticulatae Pericarpium and Citri Reticulatae Pericarpium Viride, bufothionine in Siccus Bufo and chlorogenic acid in White Chrysanthemi Flos and Lonice Raejaponicae Caulis. However, calycosin-7-glucoside and calycosin in Astragali Radix Praeparata Cum Melle and chlorogenic acid in Lonicerae Japonicae Flos, Yellow Chrysanthemi Flos and Mori Folium were all <10%, which is consistent with the CH. Decoction pieces can be stored for up to six months, but it is recommended that volatile oil-containing and animal-based decoction pieces should not be stored for more than one month. This study provides new perspectives for the stability and quality control studies of TCM. [ABSTRACT FROM AUTHOR]

Phyllostachys glauca McClure leaves (PML), as economical natural product material, contain abundant phenols, particularly flavonoids, with significant biological activities that contribute to their widespread applications in food, pharmaceutical, and cosmetic industries. To study the significant phenols in PML, an ultrasound-assisted deep eutectic solvent extraction method with high efficiency and low toxicity was established for extracting the phenols from PML and the bamboo leaves of 17 other bamboo species. Using the Ultra Performance Liquid Chromatography (UPLC) method, the content of phenols in the leaves of the 18 bamboo species was determined. PML were found to contain neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, isoorientin, and orientin at contents of 0.793 ± 00.17 mg/g, 0.491 ± 0.0081 mg/g, 0.485 ± 0.0043 mg/g, 0.465 ± 0.0021 mg/g, and 0.044 ± 0.0005 mg/g, respectively, with a total content of 2.278 mg/g. These contents were significantly higher than those found in the leaves of 17 other bamboo species. Additionally, the significant phenols, neochlorogenic acid, chlorogenic acid, and cryptochlorogenic acid were found simultaneously in the leaves of 15 bamboo species, especially in PML. Therefore, PML can be viewed as the natural product material with considerable application values, owing to its abundant phenols, and can exhibit its importance in utilizing neochlorogenic acid, chlorogenic acid, and cryptochlorogenic acid as resources. [ABSTRACT FROM AUTHOR]

Pseudotrimezia juncifolia (Klatt) Lovo & A. Gil (Iridaceae) is a popularly known species with primarily ornamental economic interest. It has traditional uses as purgative, in conditions related to the menstrual cycle, for blood purification, as wound healing, and as anti-inflammatory. The anti-inflammatory and antinociceptive activities of the decoction from its aerial stems, corms, and stamens are described here with dereplication studies on LC-MS/MS supported by the GNPS platform, where phenolic compounds were annotated and correlated with its biological activity. The decoction was evaluated in chemical (formalin and capsaicin) and thermal (hot plate) induced nociception or carrageenan-induced inflammation in mice. Decoction (at 10, 30, or 100 mg/kg doses) significantly reduced formalin- or capsaicin-induced nociception. All doses also demonstrated an antinociceptive effect in the hot plate model increasing the time the animal spent in responding to thermal signal. Naloxone partially reversed the antinociceptive effect. An anti-inflammatory effect was observed since a reduction in cell migration, protein extravasation interleukin-1, and tumor necrosis factor production induced by carrageenan in the subcutaneous air pouch was quantified. Metabolomic analyses showed a predominance of phenolic substances, mainly flavonoids and chlorogenic acids. The literature showed that these two groups have significant anti-inflammatory and analgesic activity, and chemical data corroborate the pharmacological results observed. [ABSTRACT FROM AUTHOR]

The study investigates the phytochemical profile, antioxidant capacity, and antimicrobial activities of ethanol (ChL-EtOH) and ethyl acetate (ChL-EtOAc) extracts from Chamaenerion latifolium L. (ChL) harvested in Kazakhstan. The ChL-EtOH extract exhibited higher total phenolic (267.48 ± 3.44 mg GAE/g DE) and flavonoid content (24.18 ± 1.06 mg QE/g DE) compared to ChL-EtOAc. HPLC-UV-ESI/MS identified key phenolic acids and flavonoids, including gallic acid, chlorogenic acid, and quercetin 3-glucoside. FT-IR analysis confirmed the presence of characteristic functional groups. Antioxidant assays revealed strong DPPH scavenging and FRAP activities, with ChL-EtOH showing superior results (IC50 = 21.31 ± 0.65 μg/mL and 18.13 ± 0.15 μg/mL, respectively). Additionally, ChL-EtOH displayed notable antimicrobial efficacy against Gram-positive and Gram-negative bacteria, as well as the fungal strain Candida albicans. These findings suggest that ethanol extraction is more efficient for isolating bioactive compounds from ChL, underscoring its potential for pharmaceutical and nutraceutical applications. [ABSTRACT FROM AUTHOR]

To optimize the extraction process of Eucommia ulmoides leaves by extracting the main active component chlorogenic acid, and to compare the composition and content of Eucommia ulmoides leaves from different regions. With chlorogenic acid as the target product, the contents of five active ingredients in leaves of C. intifolia and C. intifolia were determined by single factor experiment and Box-Behnken model, and the contents of rutin, pinelliol diglucoside, hyperoside, chlorogenic acid and gallic acid were determined by high performance liquid chromatography. The influence of single factor conditions on the extraction effect of both leaves was as follows: extraction temperature extraction time ethanol volume fraction> solid-liquid ratio. Under the optimal extraction conditions, the yield of chlorogenic acid was 62.94%, and that of chlorogenic acid was 47.84%. The contents of chlorogenic acid in leaves of C. sinensis were the highest. The contents of rutin and pinereol diglucoside in leaves of C. sinensis were 1. 97 and 6. 59 times higher than those of C. sinensis, but the contents of hyperoside and chlorogenic acid in leaves of C. sinensis were 1.41 and 1. 16 times higher than those of C. sinensis. This study aims to provide a theoretical basis for the selection of Eucommia species in different regions and clinical application standards and help to evaluate the quality of Eucommia ulmoides in different regions more comprehensively. [ABSTRACT FROM AUTHOR]

Consumers' demand for foods with high nutritional value and health benefits has fueled the development of prebiotic foods. In coffee industry, cherries transformation into roasted beans generates a large amount of waste/by-products (pulp/husks, mucilage, parchment, defective beans, silverskin and spent coffee grounds) that usually end up in landfills. The possibility to use coffee by-products as relevant sources of prebiotic ingredients is herein ascertained. As a prelude to this discussion, an overview of pertinent literature on prebiotic action was conducted, including on biotransformation of prebiotics, gut microbiota, and metabolites. Existing research indicates that coffee by-products contain significant levels of dietary fiber and other components that can improve gut health by stimulating beneficial bacteria in the colon, making them excellent candidates for prebiotic ingredients. Oligosaccharides from coffee by-products have lower digestibility than inulin and can be fermented by gut microbiota into functional metabolites, such as short-chain fatty acids. Depending on the concentration, melanoidins and chlorogenic acids may also have prebiotic action. Nevertheless, there is still a lack of in vivo studies to validate such findings in vitro. This review shows how coffee by-products can be interesting for the development of functional foods, contributing to sustainability, circular economy, food security, and health. Coffee by-product oligosaccharides increase short-chain fatty acid levels. Melanoidins and chlorogenic acids promote the growth of lactobacillus and bifidobacteria. Coffee by-products show prebiotic potential, but further in vivo studies are required. [ABSTRACT FROM AUTHOR]

Polyphenols, natural micronutrients derived from plants, are valued for their anti‐inflammatory and antioxidant properties. The escalating global prevalence of non‐alcoholic fatty liver disease (NAFLD) underscores its status as a chronic progressive liver condition. Furthermore, the dysregulation of gut microbiota (GM) is implicated in the onset and progression of NAFLD through the actions of metabolites such as bile acids (BAs), lipopolysaccharide (LPS), choline, and short‐chain fatty acids (SCFAs). Additionally, GM may influence the integrity of the intestinal barrier. This review aims to evaluate the potential effects of polyphenols on GM and intestinal barrier function, and their subsequent impact on NAFLD. We searched through a wide range of databases, such as Web of Science, PubMed, EMBASE, and Scopus to gather information for our non‐systematic review of English literature. GM functions and composition can be regulated by polyphenols such as chlorogenic acid, curcumin, green tea catechins, naringenin, quercetin, resveratrol, and sulforaphane. Regulating GM composition improves NAFLD by alleviating inflammation, liver fat accumulation, and liver enzymes. Furthermore, it improves serum lipid profile and gut barrier integrity. All of these components affect NAFLD through the metabolites of GM, including SCFAs, choline, LPS, and BAs. Current evidence indicates that chlorogenic acid, resveratrol, quercetin, and curcumin can modulate GM, improving intestinal barrier integrity and positively impacting NAFLD. More studies are necessary to evaluate the safety and efficacy of naringenin, sulforaphane, and catechin. [ABSTRACT FROM AUTHOR]

Postoperative abdominal adhesion (PAA) widely occurs after abdominal surgery, which often produces severe complications. However, there were still no satisfactory anti-adhesive products including barriers and anti-adhesive agents. Herein, we developed a ROS-responsive and scavenging hydrogel barrier, termed AHBC/PSC, wherein the monomer AHBC was synthesized by phenylboronic acid (PBA)-modified hyaluronic acid (HA-PBA) further grafted with adipic dihydrazide (ADH) and PBA-based chlorogenic acid (CGA) via ROS-sensitive borate ester bond, and the other monomer PSC was constructed by polyvinyl alcohol (PVA) grafted with sulfated betaine (SB) and p-hydroxybenzaldehyde (CHO). Further, the double crosslinked AHBC/PSC hydrogel was successfully fabricated between AHBC and PSC via forming dynamic covalent acylhydrazone bonds and borate ester bonds. Results showed that AHBC/PSC hydrogel had in situ gelation behavior, satisfactory mechanical properties (storage modulus of about 1 kPa and loss factor Tan δ of about 0.5), suitable wet tissue adhesion strength of about 2.3 kPa on rat abdominal wall, and good biocompatibility, achieving an ideal physical barrier. Particularly, CGA could be responsively released from the hydrogel by breakage of borate ester bonds between CGA and PBA based on high reactive oxygen species (ROS) levels of damaged tissue and exhibited great ROS scavenging capability to regulate inflammation and promote the polarization of macrophages from pro-inflammatory M1 phenotype to anti-inflammatory M2 phenotype. Moreover, the grafted SB as a zwitterionic group could reduce protein adsorption and fibroblast adhesion. Finally, the in vivo experiments revealed that AHBC/PSC hydrogel with good safety and in vivo retention behavior of about 2 weeks, effectively prevented PAA by regulating the inflammatory microenvironment and alleviating the fibrosis process. In brief, the versatile AHBC/PSC hydrogel would provide a more convenient and efficient approach for PAA prevention. Postoperative abdominal adhesion (PAA) widely occurs after surgery and is often accompanied by severe complications. Excessive inflammation and oxidative stress are very crucial for PAA formation. This study provides a ROS-responsive and scavenging hydrogel with suitable mechanical properties, good biocompatibility and biodegradability, and resistance to protein and fibroblast. The antioxidant and anti-inflammatory active ingredient could be responsively released from the hydrogel via triggering by the high ROS levels in the postoperative microenvironment thereby regulating the inflammatory balance. Finally, the hydrogel would effectively regulate the development process of PAA thereby achieving non-adhesion wound healing. [Display omitted] [ABSTRACT FROM AUTHOR]

Background: Coffee leaves are a major source of bioactive components and are used as ethnomedicine. However, despite their traditional medicinal use, information about their effects on antihyperlipidemia remains limited. Methods: The aims of this study were to evaluate the main components of leaf extracts from Arabica and Robusta coffees and to examine the potential of these coffee leaves in reducing lipid digestion and absorption in vitro. Results: Coffee leaf extracts from Arabica coffee contain a high amount of caffeine, whereas extracts from Robusta coffee contain high amounts of chlorogenic acid (CGA) and caffeine. Additionally, leaf extracts from Arabica and Robusta coffee demonstrated the inhibition of pancreatic lipase, decreased micellar cholesterol solubility, and reduced bile acid binding. Furthermore, these extracts resulted in a reduction in cholesterol uptake in Caco-2 cells. Molecular docking experiments supported this discovery, showing CGA and caffeine binding to Niemann–Pick C1-like 1 (NPC1L1), a key protein in cholesterol absorption. The results indicated that CGA and caffeine can competitively bind to NPC1L1 at the cholesterol binding pocket, reducing its cholesterol binding rate. These findings suggest that coffee leaves might help suppress lipid absorption and digestion, highlighting their potential use in preventing and treating hyperlipidemia. [ABSTRACT FROM AUTHOR]

The antioxidant activities of lycopene (LY), lutein (LU), chlorogenic acid (CA), and delphinidin (DP) were tested in vitro on H9c2 cell-based models. Some indicators, such as the generation of reactive oxygen (ROS), the quantification of cell antioxidant activity (CAA), and the expressions of SOD, GSH-Px, and CAT, were calculated to examine their antioxidant interactions. From our results, the phytochemical mixtures (M1: CA-LU: F3/10, M2: DP-CA: F7/10, M3: DP-LY: F5/10) displayed strong synergistic effects based on the generation of ROS and the quantification of CAA. However, great antagonistic bioactivities were seen in the combinations of LY-LU: F5/10 (M4), CA-LU: F9/10 (M5), and DP-LY: F7/10 (M6). Western blotting analysis indicated that the possible mechanism underlying the synergistic antioxidant interactions among phytochemical combinations was to enhance the accumulation of Nrf2 in the nucleus and the expression of its downstream antioxidant enzymes, HO-1 and GCLC. The combinations (M1–M3 groups) showed significant protection against the loss of mitochondrial membrane potential than individual groups to avoid excessive ROS production. The M4–M6 groups exerted antagonistic protective effects compared with the individual groups. In addition, lutein and lycopene absorption was improved more because of the presence of chlorogenic acid and delphinidin in the M1 and M3 groups, respectively. However, delphinidin significantly reduced the cellular uptake of lycopene in the M6 group. It appeared that antioxidant interactions of phytochemical combinations may contribute to the restoration of cellular redox homeostasis and lead to an improvement in diet quality and collocation. [ABSTRACT FROM AUTHOR]

Antispasmodic agents are crucial in managing gastrointestinal motility disorders by modulating muscle contractions and reducing symptoms like cramping and diarrhea. This study investigated the antispasmodic potential of different coffee bean extracts, including light coffee (LC), medium coffee (MC), and dark coffee (DC), on ileum contractions induced by potassium chloride (KCl), and elucidated their mechanisms of action using in vitro isolated tissue techniques. The results demonstrated that all coffee extracts reduced spontaneous contractions of rat ileum tissue in a dose-dependent manner. Among these, LC showed the most significant reduction in ileum contractions, particularly at higher concentrations. The key findings reveal that LC at 5 mg/mL significantly reduced CaCl2-induced contractions in isolated rat ileum tissue, indicating that LC may inhibit calcium influx or interfere with calcium signaling pathways. The presence of nifedipine, propranolol, and N-nitro-L-arginine methyl ester (L-NAME) have been confirmed in their involvement; they block calcium influx and calcium channels and activate β-adrenergic pathways as part of LC's mechanism of action. The presence of their active compounds, particularly chlorogenic acid and caffeine, likely contributes to the observed antispasmodic effects. These findings suggest that LC exerts its antispasmodic effects by targeting key mechanisms involved in muscle spasms and intestinal motility, providing a potential for managing such conditions. [ABSTRACT FROM AUTHOR]