Trolox Equivalent Antioxidant Capacity Research Articles

Metabólito secundário do extrato de folhas de cardamomo de Java em função de sombreamento e doses de nitrogênio

ABSTRACT Java cardamom is a species of herbal medicinal plant endemic to West Java, Indonesia, that has been used for a long time in traditional medicine. The effects of shading and nitrogen doses on the Gas Chromatography-Mass Spectrometry (GC-MS) profile, total phenolic and flavonoid contents, and antioxidant activity in the ethanol extract of Java cardamom leaves were investigated in this study. The experiment was conducted using a split-plot scheme with three replicates. The main plot constituted shading (without, 25, 50, and 75%), and the sub-plot comprised nitrogen doses (control, 0.9, and 1.36 g nitrogen fertilizer per polybag). GC-MS analysis found 27 compounds in the ethanol extract of Java cardamom leaves. The compounds 1.8-Cineole (36.87%) and 5-Hydroxy-3,7,4’-trimethoxyflavone (18.83%) had the highest concentrations in the combination of 25% shade with 0.9 g N fertilizer per polybag and 50% shade with 1.36 g N fertilizer per polybag. The highest total phenolic content (1.26 mg gallic acid equivalent g-1 dry weight), flavonoid content (3.42 mg quercetin equivalent g-1 dry weight), and ferric reducing antioxidant power (FRAP) antioxidant activity (5.80 μmol Trolox equivalent antioxidant capacity g-1 dry weight) were found with 75% shade. Metabolites 1-Methyl-4-isopropyl-cis-3-hydroxycyclohex-1-ene-6-one, 4-propoxy-catechol, cyclohexane, tert-pentyl-, cis-p-Menth-2,8-dienol, cis-carveol, and cis-p-mentha-1(7),8-dien-2-ol were responsible for antioxidant activity in the ethanol extract of Java cardamom leaves. Shade of 75% is recommended to produce optimal antioxidant activity and phenolic and flavonoid content from Java cardamom leaf extract.

Kuşburnu (Rosa canina L.) Tohumu Tozunun Mineral ve Biyoaktif Bileşen İçerikleri

This study determined the bioactive components of rosehip (Rosa canina L.) extract obtained via supercritical carbon dioxide extraction. The total phenolic content of its extract was 214.4 mg gallic acid equivalent/kg, with the total flavonoid content of 21.1 mg quercetin equivalent/kg. The antioxidant activity of the extract, which was determined by the DPPH (2,2-diphenyl-1-picrylhydrazyl) assay, was 64.8 μmol Trolox® equivalent antioxidant capacity (TEAC)/g. Gas chromatography-mass spectrometry identified 15 bioactive components in the extract. Additionally, pre- and post-processing heavy metal analyses were conducted on rosehip powder and seeds using inductively coupled plasma mass spectrometry (ICP-MS). Results showed that metal concentrations ranged from 0.064 to 9134.1 mg/kg in rosehip powder and from 0.143 to 1929.5 mg/kg in seeds, with the concentrations of potassium and magnesium as the highest. Despite the limited uses of rosehip products, this study indicated that wild rosehips are rich in functional components with potential health benefits.

Fatty acid profile, total phenolic content and antioxidant capacity of seaweeds in Salinas Bay, Ecuador

The coastal waters of Ecuador have emerged as a favorable environment for the growth and development of several species of edible macroalgae or seaweeds. In this study, fatty acid profiles, total soluble polyphenol content, and antioxidant capacity of seven species of macroalgae (four red algae, two brown algae and one green algae) collected from Salinas Bay, Santa Elena Province, Ecuador, were evaluated. Saturated fatty acids C14:0, C15:0, C16:0 and C18:0 were found in almost all organisms, from which palmitic acid showed the highest proportion (10.7-37.1%). Green and brown seaweeds exhibited a major content of unsaturated fatty acids. Monounsaturated fatty acid composition was represented by several isomers of C16:1 and C18:1, mainly palmitoleic (3.0-6.6%) and oleic (0.8-11.1%) acids. Polyunsaturated fatty acid composition ranged from 5.47 to 26.61%, with ratios of total n-6/n-3 acids lower than 3.3. Linoleic (C18:2, n-6, 1.16-7.73%) and eicosapentaenoic (C22:5, EPA, n-3, 1.44-5.92%) acids were found in the majority of the seaweeds. The total polyphenol content of the seaweeds was determined by the Folin-Ciocalteu method (765 nm), and ranged from 147.28 to 1,062.20 mg equivalents of gallic acid (GAE) per 100 g of dry seaweed. The antioxidant capacity was determined by the Trolox equivalent antioxidant capacity (517 nm). Only Acanthophora spicifera, Spatoglossum schroederi, and Ulva lactuca showed good radical-scavenging (14.18-3,379.47 mg Trolox equivalents (TE) per 100 g of dry seaweed). The results confirm the potential use of macroalgae in the study area as an important source of nutritional compounds, especially brown and green algae, which could be a recommended food supplement even in the treatment of some diseases.

Determination of cytotoxic, antioxidant activities and LC/MS-MS profiles of three endemic Verbascum L. species

The Verbascum genus includes many species used in folk medicine or the treatment of various diseases. In this study, the cytotoxic, antioxidant activity, and LC/MS-MS profiles of three Verbascum species, which are endemic in Eskişehir and its surroundings, were investigated. The cytotoxic effects of methanol extract of V.detersile, V. eskisehirensis, and V.gypsicola species on the cervical (HeLa) and ovarian cancer (SKOV-3) cells were investigated using a colorimetric assay. The results indicated that cytotoxic effect was not observed after treatment of SKOV-3 cells with Verbascum. On the other hand, the cytotoxic activity of V. detersile was found to be 0.1910 mg/dL and 1.057 mg/dL for HeLa cells after 24 or 48 hours incubation with V.detersile, respectively. The antioxidant activity was determined as 1,1-diphenyl-2-picrylhydrazyl (DPPH•) radical scavenging activity, trolox equivalent antioxidant capacity (TEAC assay), and also the total phenolic content of the samples was found. Total phenols were estimated as Gallic acid equivalents (GAE), expressed as mg Gallic acid/in 1g extract previously described by Singletton. LC/MS profiles separation and detection of phytochemicals of the extract were performed on a Shimadzu UPLC system consisting of a vacuum degasser, an autosampler (SIL20A Shimadzu Autosampler), a binary pump (LC20AD Shimadzu), an oven (CTO20A Shimadzu Column Oven) and DAD dedector (SPD M20A Shimadzu DAD Detector). In terms of antioxidant activity, V. gypsicola was found to have the least antioxidant activity among the three extracts, which is also correlated with the total amount of phenolic content in its content. In this way, it differs from other species. V. detersile exhibits a different chemical profile from the other two species with the iridoid catalpol derivatives it contains. Apigenin pentoside is the only flavonoid molecule detected in V. detersille.

Tropical Cultivated Hericium erinaceus: Antioxidant Capacities of Heat Treatment on Fresh and Oven-Dried Fruiting Bodies

ABSTRACT Consumption of mushrooms in daily diet has become tremendously popular. This study was carried out to investigate the antioxidant capacities (DPPH radical scavenging, reducing power, inhibition of lipid peroxidation, and trolox equivalent antioxidant capacity) and total phenolic content on the heat treatments (boiling, steaming, microwave and pressure cooking) of fresh and oven-dried fruit body of Hericium erinaceus cultivated in tropical region. Samples exposed to 5 minutes to both steaming and pressure-cooking exhibited a significant increase in DPPH and reducing powers activities. The TEAC was also increased significantly in 5 minutes of both microwaving and pressure-cooking samples. Most of the cooked samples showed a high phenolic content. Weak correlation between the antioxidant activities and total phenolic content of samples indicates the antioxidant activity is not due to the amount of phenolic compounds. Thus, steaming and pressure-cooking might be the best cooking method to maintain high antioxidant content of H. erinaceus.

Ultra-High-Resolution Liquid Chromatography Coupled with Electrospray Ionization Quadrupole Time-of-Flight Mass Spectrometry Analysis of Tessaria absinthioides (Hook. & Arn.) DC. (Asteraceae) and Antioxidant and Hypocholesterolemic Properties.

Recently, we reported the chemical profile and the hypocholesterolemic effects of a decoction of Tessaria absinthioides (Hook. & Arn.) DC. (Asteraceae). In this study, we evaluated a methanolic extract (METa) instead. Metabolite profiling was conducted using ultra-high-resolution liquid chromatography coupled with electrospray ionization quadrupole time-of-flight mass spectrometry (UHPLC-ESI-QTOF-MS), identifying thirty compounds, including flavonoids, phenolic acids, fatty acids, and phorbolesters. Antioxidant properties were assessed through 2,2-diphenyl-1-picrylhydrazyl (DPPH), Trolox equivalent antioxidant activity (TEAC), ferric-reducing antioxidant power (FRAP), and inhibition of lipid peroxidation in erythrocytes (ILP) assays, exhibiting robust antioxidant activity. The in vivo impact of METa on serum lipid parameters and liver X receptors (LXRs) was evaluated in a hypercholesterolemic animal model. After 14 days on a high-fat diet, male rats received either a vehicle (V) or METa100, METa200 or METa500 (100; 200 and 500 mg METa/kg animal, respectively) for an additional two weeks. METa500 reduced total cholesterol levels (17.62%; p < 0.05) and all doses increased high-density lipoprotein cholesterol levels (METa100: 86.27%; METa200: 48.37%, and METa500: 29.42%; p < 0.0001). However, METa did not alter LXRs expression. The observed antioxidant and hypocholesterolemic properties of METa may be linked to the presence of six di-caffeoylquinic acids. These findings underscore T. absinthioides as a potential candidate for the treatment of metabolic disease.

Characterization of the Antiproliferative Activity of Sargassum muticum Low and High Molecular Weight Polysaccharide Fractions.

The extract obtained by pressurized hot water extraction from Sargassum muticum, to recover the bioactive compound known as fucoidan, was fractionated using membranes of 100, 50, 30, 10, and 5 kDa, obtaining five retentates and the final permeate. These fractions were characterized for phloroglucinol content, protein content, sulfate content, and trolox equivalent antioxidant capacity (TEAC); apart from oligosaccharides, FTIR and molar mass distribution were also evaluated. Retentates of 100 and 50 kDa showed higher values for phloroglucinol, TEAC, and sulfate content. The rheology of the alginate fraction was also evaluated. Regarding the potential antitumoral activity, all fractions were assessed in MCF-7 cells using a metabolic activity assay based on the reduction of a tetrazolium compound, the most efficient being R100 and R50. Based on the results, these fractions were compared with commercial fucoidans at the same concentrations, and similar results were found. In addition, synergistic cytotoxic effects using two drugs commonly used in breast cancer, cis-Platinum (cis-Pt) and 5-fluorouracil (5-FU), were tested in combination with R100 and R50. Promising results were obtained when the retentate and the drugs were mixed, showing an improvement in the cytotoxicity induced by the chemotherapy.

Assaying the Use of a Leonardite-Suspension Concentrate-Based Product as a Potential Biostimulant to Enhance Growth, NPK Use Efficiency, and Antioxidant Capacity in Lactuca sativa L.

Biostimulants are presented as a potential tool to achieve the main objectives of modern agriculture: increase crop yield and nutritional quality while reducing chemical fertilizer use. Here, we investigated the use of a HS-based product (BLACKJAK®, from Sofbey S.A., Mendrisio, Switzerland) as a biostimulant to enhance plant growth, nitrogen (N), phosphorus (P), and potassium (K) use efficiency, as well as antioxidant capacity. For this purpose, BLACKJAK® was applied to lettuce (Lactuca sativa L.) as radicular (‘R’) and foliar (‘F’) at doses: 0.20 mL/L (R1), 0.40 mL/L (R2), 0.60 mL/L (R3), and 0.80 mL/L (R4), 5.00 mL/L (F1), 7.50 mL/L (F2), 10.00 mL/L (F3), and 12.50 mL/L (F4), along with a control. Shoot fresh weight (FW) and dry weight (DW), leaf area, NPK use efficiency parameters, and antioxidant capacity were evaluated. Our results showed that R1, R2, R3, F2, and F3 enhanced shoot FW and leaf area, while only R3 increased shoot DW. Furthermore, in general, most of the doses employed enhanced NPK use efficiency parameters such as apparent crop recovery, nutrient export, physiological efficiency, and internal utilization of applied nutrients. Similarly, HS also increased ascorbate, glutathione, and phenol concentrations, showing an improvement in antioxidant capacity measured through FRAP (Ferric Reducing Antioxidant Power) and TEAC (Trolox Equivalent Antioxidant Capacity) assays. Overall, root-HS offered better results than foliar-HS, especially at R3. Hence, our results indicate that BLACKJAK® could be a good candidate to enhance crop productivity and nutritional quality while reducing the use of chemical NPK fertilizers.

Antioxidant Response, Phenolic Compounds and Yield of Solanum tuberosum Tubers Inoculated with Arbuscular Mycorrhizal Fungi and Growing under Water Stress.

Solanum tuberosum (potato) is one of the most common crops worldwide; however, it is sensitive to water stress, which necessitates the identification of alternative tools to improve their production. Here, we evaluated the inoculation of two arbuscular mycorrhizal fungi (AMF) strains, Claroideoglomus claroideum (CC), Claroideoglomus lamellosum (HMC26), and the MIX (CC + HMC26) in yield and phenolic and antioxidant response using chromatographic and spectroscopic methods in potato crops, at increasing levels of water stress, namely, with 100% (0), 70% (S1), and 40% (S2) soil humidity. Two caffeoylquinic acid isomers were detected and their levels showed a tendency to increase under stress together with the AMF inoculation, reaching up to 19.2 mg kg-1 of 5-caffeoylquinic acid and 7.4 mg kg-1 of caffeoylquinic acid isomer when CC was inoculated, and potato plants grew at the highest water starvation condition (S2). Regarding antioxidant activities, a differentiated response was detected depending on the AMF strain, highlighting the effect of HMC26 on Trolox equivalent antioxidant capacity (TEAC) method and CC in cupric reducing antioxidant capacity (CUPRAC) method, reaching up to 1.5 μmol g-1 of TEAC in plants inoculated with HMC26 and 0.9 μmol g-1 of CUPRAC in plants inoculated with CC, both in potato tubers of plants growing under the S2 stress condition. Meanwhile, the use of AMF did not influence the number and biomass of the tubers, but significant changes in the biochemical properties of tubers were observed. The results suggest that specific AMF adaptations to water stress must be considered when inoculation procedures are planned to improve the yield and quality of tubers in potato crops.

Blockade of angiotensin II modulates insulin-like growth factor 1-mediated skeletal muscle homeostasis in experimental steatohepatitis

Sarcopenia is associated with mortality in patients with nonalcoholic steatohepatitis (NASH). Angiotensin II receptor blocker (ARB) has been suggested to prevent sarcopenia, but reports on its effect on NASH-derived skeletal muscle atrophy in conjunction with insulin-like growth factor 1 (IGF-1)-mediated muscle homeostasis are few. Our aim was to examine the combined effect of the ARB losartan and IGF-1 replacement on skeletal muscle atrophy in a methionine–choline deficient (MCD) diet-fed murine steatohepatitis model. The MCD-fed mice developed steatohepatitis and skeletal muscle atrophy, as indicated by the reduction of psoas muscle mass and attenuation of forelimb and hindlimb grip strength. Significantly suppressed steatohepatitis and skeletal muscle atrophy was observed after single treatment with ARB or IGF-1, and these effects were augmented after combination treatment. Treatment with ARB and IGF-1 effectively inhibited ubiquitin proteasome-mediated protein degradation by reducing forkhead box protein O1 (FOXO1) and FOXO3a transcriptional activity in the skeletal muscle. Combined ARB and IGF-1 decreased the intramuscular expression of proinflammatory cytokines (i.e., TNFα, IL6, and IL1β) and increased the Trolox equivalent antioxidant capacity and antioxidant enzymes (CAT, GPX1, SOD2, and CYTB). This antioxidant effect was based on downregulation of NADPH oxidase (NOX) 2, normalization of mitochondrial biogenesis and dynamics. Moreover, ARB increased the hepatic and plasma IGF-1 levels and improved steatohepatitis, leading to enhanced skeletal muscle protein synthesis mediated by IGF-1/ AKT/ mechanistic target of rapamycin signaling. Collectively, combined ARB and IGF-1 replacement could be a promising new therapeutic target for NASH-derived skeletal muscle wasting.

Production of highly antioxidant lignin nanoparticles from a hardwood technical lignin

Eucalypt kraft lignin isolated in a LignoBoost™ pilot plant was characterized by GC–MS, ICP-OES, DSC, HPSEC, 31P NMR, and HSQC 2D-NMR to be used without any further processing to produce lignin nanoparticles (LNPs) by nanoprecipitation. Tetrahydrofuran (THF) was used as a solvent, and water as a non-solvent. Microscopic analysis (TEM) showed that LNPs were regularly spherical with some hollow particles dispersed in-between, and sizes were tunable by changing the solvent dripping rate onto the non-solvent. LNP particle sizes had a bimodal distribution, with the largest population having an average apparent hydrodynamic diameter ranging from 105.6 to 75.6 nm. Colloidal dispersions of LNPs in water presented good stability in different dilutions without significant size changes upon storage at pH close to neutral for as long as 45 days. Zeta potentials around −40 mV were obtained for LNP suspensions at pH ranging from 7 to 9. The high carbohydrate content (circa 10 % on a dry basis, mostly xylans) of the lignin precursor did not interfere in LNP formation, whose antioxidant activity was expressive as demonstrated by the ABTS assay at pH 7.4, with an EC50 of 4.04 μg mL−1. Also, the Trolox® equivalent antioxidant capacity (TEAC) of LNPs reached 1.90 after 40 min reaction time.

Zuccagnia punctata Cav., a Potential Environmentally Friendly and Sustainable Bionematicide for the Control of Argentinean Horticultural Crops.

This research was designed to investigate the metabolite profiling, phenolics, and flavonoids content as well as the potential nematicidal properties of decoction (ZpDe), orange-yellow resin (ZpRe) and essential oil (ZpEO) from Argentinean medicinal plant Zuccagnia punctata Cav. Additionally, the antioxidant and antibacterial properties of ZpDe and ZpEO were determined. Metabolite profiling was obtained by an ultrahigh-resolution liquid chromatography MS analysis (UHPLC-ESI-QTOF/OT-MS-MS) and GCMS. The nematicidal activity was assayed by a standardized method against Meloidogyne incognita. The antioxidant properties were screened by four methods: (2,2-diphenyl-1-picrylhydrazyl assay (DPPH), Trolox equivalent antioxidant activity assay (TEAC), ferric-reducing antioxidant power assay (FRAP), and lipid peroxidation in erythrocytes (ILP). The antibacterial activity was evaluated according to the Clinical and Laboratory Standards Institute (CLSI) rules. The ZpDe, ZpRe and ZpEO displayed a strong nematicidal activity with an LC50 of 0.208, 0.017 and 0.142 mg/mL, respectively. On the other hand, the ZpDe showed a strong DPPH scavenging activity (IC50 = 28.54 µg/mL); ILP of 87.75% at 250 µg ZpDe/mL and moderated antimicrobial activity. The ZpEO showed promising activity against a panel of yeasts Candida albicans and non-albicans (ATCC and clinically isolated) with MIC values from 750 to 1500 µg/mL. The ZpDe showed a content of phenolics and flavonoid compounds of 241 mg GAE/g and 10 mg EQ/g, respectively. Fifty phenolic compounds were identified in ZpDe by ultrahigh-resolution liquid chromatography (UHPLC-PDA- Q-TOF-MS) analysis, while forty-six phenolic compounds were identified in ZpRe by UHPLC-ESI-Q-OT-MS-MS and twenty-nine in ZpEO using a GC-MS analysis, updating the knowledge on the chemical profile of this species. The results support and standardize this medicinal plant mainly as a potential environmentally friendly and sustainable bionematicide for the control of Argentinean horticultural crops including tomatoes and peppers and as a source of antimicrobial and antioxidant compounds which could be further explored and exploited for potential applications.

Saliva Sampling Material Matters: Effects on the Results of Saliva Analysis in Pigs.

The use of saliva as a biological sample from pigs is of high practical interest because blood collection from pigs is difficult and stressful. In this study, the influence of two different materials, a cotton roll and a polypropylene sponge, in porcine saliva collection was evaluated. For this purpose, the effect of the material used for sampling was evaluated in a panel of 13 analytes, including those related to stress (cortisol and oxytocin), inflammation and immunity (adenosine deaminase, haptoglobin and myeloperoxidase), redox homeostasis (the cupric reducing ability of saliva, the ferric reducing activity of saliva, and the Trolox equivalent antioxidant capacity), and sepsis (procalcitonin), as well as other routine analytes related to metabolism and different tissues and organs, such as lactate dehydrogenase, creatine kinase, urea, and total protein concentration. The polypropylene sponge provided a higher sample volume than the cotton roll. Although the results of some salivary analytes were equivalent for both materials, other analytes, such as creatine kinase, haptoglobin and total proteins, showed significant differences depending on the material used for saliva collection. Therefore, the type of material used for salivary collection in pigs should be considered when interpreting the results of analyses of the salivary analytes.

Antioxidant peptides obtained from abalone muscle hot water extract during in‐vitro simulated digestion and Caco‐2 cell absorption

SummaryAfter abalone muscle hot water extract (AHE) was digested in‐vitro, three components (F1, F2, and F3) were separated using the Sephadex G‐15 column, and the antioxidant activity and absorption across Caco‐2 cells were analysed. The result showed that the half‐maximal inhibitory concentrations of F2 for scavenging hydroxyl and ABTS radicals were 0.763 and 0.028 mg mL−1, respectively, which were lower than those of F1 and F3. Moreover, the Trolox equivalent antioxidant capacity was increased after the F2 was transported through the Caco‐2 cell‐monolayered Transwell chamber. The mass spectrometry analysis revealed that the transported samples contained oligopeptides (Leu‐Tyr, Gly‐Ala‐Ala, and His‐Gly‐Ser‐Ala), and the Leu‐Tyr displayed the highest antioxidant activity and transport rate. Overall, these results suggest that oligopeptides with high antioxidant capacities can be produced from AHE through a combination of in‐vitro digestion and absorption across Caco‐2 cells.

Biocosmetics Made with Saccharina latissima Fractions from Sustainable Treatment: Physicochemical and Thermorheological Features.

This work deals with the formulation of natural cosmetics enriched with antioxidant fractions from the ultrasound treatment (US) of the brown seaweed Saccharina latissima. The challenge was the development of a cosmetic matrix without jeopardizing the thermorheological features of the creams, adding microparticles containing the antioxidant fractions using two different carriers, mannitol and alginate. The fundamental chemical characteristics of seaweed and the extracts obtained via sonication, as well as the antioxidant properties of the latter, were analyzed. The highest TEAC (Trolox equivalent antioxidant capacity) value was identified for the extracts subjected to the longest processing time using ultrasound-assisted extraction (240 min). A similar yield of microparticle formulation (around 60%) and load capacity (about 85%) were identified with mannitol and alginate as carriers. Color testing of the creams exhibited small total color differences. The rheological results indicated that the testing temperature, from 5 to 45 °C, notably influenced the apparent viscosity of the matrices. All creams were adequately fitted with the two parameters of the Ostwald-de Waele model, with the flow consistency index following an Arrhenius dependency with the testing temperature. Neither hysteresis nor water syneresis was observed in the proposed cosmetics during 6 months of cold storage at 4-6 °C.

The Chemical Profile, Antioxidant, and Anti-Lipid Droplet Activity of Fluid Extracts from Romanian Cultivars of Haskap Berries, Bitter Cherries, and Red Grape Pomace for the Management of Liver Steatosis

This study aimed to investigate, for the first time, the chemical composition and antioxidant activity of fluid extracts obtained from three Romanian cultivars of haskap berries (Lonicera caerulea L.) var. Loni, bitter cherries (Prunus avium var. sylvestris Ser.) var. Silva, and pomace from red grapes (Vitis vinifera L.) var. Mamaia, and their capacity to modulate in vitro steatosis, in view of developing novel anti-obesity products. Total phenolic, flavonoid, anthocyanin, and ascorbic acid content of fluid extracts was spectrophotometrically assessed and their free radical scavenging capacity was evaluated using Trolox Equivalent Antioxidant Capacity (TEAC) and free 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical inhibition assays. The Pearson coefficients showed a moderate correlation between the antioxidant activity of fluid extracts and their phenolic content, but a strong correlation between anthocyanin and ascorbic acid content. HPLC analysis identified and quantified the main phenolic compounds of chlorogenic and syringic acid, catechin, and glycosylated kaempferol, apigenin, and quercetin, in variable proportions. An in vitro experimental model of steatosis was developed in HepG2 hepatocytes treated with a mixture of free fatty acids. Cell culture analyses showed that cytocompatible concentrations of fluid extracts could significantly reduce the lipid accumulation and inhibit the reactive oxygen species, malondialdehyde, and nitric oxide secretion in stressed hepatocytes. In conclusion, these results put an emphasis on the chemical compounds’ high antioxidant and liver protection capacity of unstudied fluid extracts obtained from Romanian cultivars of bitter cherries var. Silva and pomace of red grapes var. Mamaia, similar to the fluid extract of haskap berries var. Loni, in particular, the positive modulation of fat deposition next to oxidative stress and the lipid peroxidation process triggered by fatty acids in HepG2 hepatocytes. Consequently, this study indicated that these fluid extracts could be further exploited as hepatoprotective agents in liver steatosis, which provides a basis for the further development of novel extract mixtures with synergistic activity as anti-obesity products.

Gastrointestinal Effects on the Antioxidant and Immunomodulatory Properties of South African Fynbos Honey.

The Fynbos biome, Western Cape Province, South Africa, produces a unique honey from Apis mellifera capensis. The bioactivity of Fynbos (FB1-FB6) honeys and Manuka, unique manuka factor 15+ (MAN UMF15+) honey subjected to simulated in vitro digestion, was compared. The effect of each phase of digestion on the antioxidant properties and nitric oxide- (NO-) associated immunomodulatory effects was determined. The total phenolic content of MAN (UMF15+) was higher than that of FB honeys, and following digestion, the percentage bioaccessibility (BA) was 68.6% and 87.1 ± 27.0%, respectively. With the Trolox equivalent antioxidant capacity assay, the activity of FB1 and FB6 was similar to MAN (UMF15+) but reduced for FB2, FB3, FB4, and FB5 with a %BA of 77.9% for MAN (UMF15+) and 78.2 ± 13.4% for FB. The oxygen radical absorbance capacity of MAN (UMF15+) and FB honeys was similar and unaltered with digestion. In a cellular environment, using colon adenocarcinoma (Caco-2) cells, both undigested and the gastric digested honey reduced 2,2'-azobis-(2-amidinopropane) dihydrochloride- (AAPH-) mediated peroxyl radical formation. In contrast, following gastroduodenal digestion, the formation of reactive oxygen species (ROS) was increased. In murine macrophage (RAW 264.7) cells, all honeys induced different levels of NO which was significantly increased with digestion for MAN (UMF15+) and FB1. In LPS/IFN-γ stimulated RAW 264.7 macrophages, only undigested MAN (UMF15+) effectively reduced NO levels, and with digestion, NO scavenging activity of MAN (UMF15+) was reduced but increased for FB5 and FB6. In a noncellular environment, MAN (UMF15+), FB1, FB2, and FB6 scavenged NO, and with digestion, this activity was maintained. This study has identified that undigested and gastric-digested FB honey has antioxidant properties with strong potential anticancer effects following gastroduodenal digestion, related to ROS formation. MAN (UMF15+) had anti-inflammatory effects which were lost postdigestion, and in contrast, FB5 and FB6 had anti-inflammatory effects postdigestion.

Establishment of the mechanism of purification and levigation of green chemistry-assisted biocomposites of red ochre (Gairika): synthesis, characterization, and antibacterial, prebiotic, antioxidant, and antacid activities of the traditional Ayurvedic medicine Laghu Sutashekhara Rasa

Gairika (red ochre) has a long history of influencing human civilization. Gairika is a rich source of nutrients used for reproductive and brain health. Gairika is mentioned as an antacid drug in Indian Ayurvedic medicine under Laghu Sutashekhara Rasa (LSR). However, a detailed study on LSR has not been reported to date. In the present study, LSR was prepared, and a pharmaceutical SOP (standardization procedure) was reported to obtain batch-to-batch reproducibility. LSR was characterized using FTIR, XRD, SEM-EDX, and TGA analyses. LSR was tested in vitro for its antacid activity. Advanced instrumentation revealed that LSR formation produced symmetrical particles (5–8 µm) with kaolin, kaolinite, quartz, goethite, and hematite, along with the phytoconstituents of Goghrita (clarified cow’s butter), Shunthi, and Nagawalli, as confirmed by GC-MS/MS analysis. The FTIR study revealed the formation of a chelating complex of goethite and hematite along with their phytoconstituents. XRD analysis confirmed the presence of kaolin, kaolinite, quartz, goethite, and hematite. Using in vitro antacid experiments, LSR and Shunthi demonstrated significant antacid activity as compared to antacid drugs and standards in the market, such as CaCO3. The DPPH assay revealed IC50 values of 12.16 ± 1.23 mg/mL, which is 0.0029 of Trolox-equivalent antioxidant activity. The inhibition (18 ± 4 mm) against pathogens (S. aureus, E. coli, P. aeruginosa, and B. subtilis) and the prominent growth of gut microbiota-supported strains (S. boulardii, L. paracasei, and L. plantarum) observed on LSR formulation were indicative of LSR application as a prebiotic. Here, the mechanism of purification and levigation mentioned in the classical literature of LSR was established. Overall, purification of Gairika with cow ghee and levigation with Nagawalli may enhance the solubility, bioavailability, and shelf-life of LSR through hydration and co-crystallization mechanisms. This is the first comprehensive report on the pharmaceutical validation of LSR and its characterization. The results of the present study could contribute to the development and reliable reproduction of LSR and the utility of environmental red ochre as a medicine in combination with Shunthi (Zingiber officinale Roxb.), as prescribed under Indian Ayurvedic medicine.

Oxidative Stress in Xenograft Mouse Model Exposed to Dendrimers Decorated Polydopamine Nanoparticles and Targeted Chemo- and Photothermal Therapy

Polydopamine (PDA)-based nanostructures are used for biomedical purposes. A hybrid drug nanocarrier based on a PDA decorated with polyamidoamine (PAMAM) dendrimers G 3.0 (DG3) followed by a connection with glycol (PEG) moieties, folic acid (FA), and drug doxorubicin (DOX) was used for combined chemo- and photothermal therapy (CT-PTT) of liver cancer. Oxidative stress plays a crucial role in the development of cancer, and PDA seems to have the ability to both donate and accept electrons. We investigated oxidative stress in organs by evaluating oxidative stress markers in vivo. In the liver, the level of reduced glutathione (GSH) was lower and the level of Trolox equivalent antioxidant capacity (TEAC) was higher in the group receiving doxorubicin encapsulated in PDA nanoparticles with phototherapy (PDA@DG3@PEG@FA@DOX + PTT) compared to the control group. The concentration of thiobarbituric acid reactive substances (TBARS) in livers, was higher in the group receiving PDA coated with PAMAM dendrimers and functionalized with PEG and FA (PDA@DG3@PEG@FA) than in other groups. Markers in the brain also showed lower levels of GSH in the PDA@DG3@PEG@FA group than in the control group. Markers of oxidative stress indicated changes in the organs of animals receiving PDA nanoparticles with PAMAM dendrimers functionalized with FA in CT-PTT of liver cancer under in vivo conditions. Our work will provide insights into oxidative stress, which can be an indicator of the toxic potential of PDA nanoparticles and provide new strategies to improve existing therapies.

A Commercial Clay-Based Material as a Carrier for Targeted Lysozyme Delivery in Animal Feed.

The controlled supply of bioactive molecules is a subject of debate in animal nutrition. The release of bioactive molecules in the target organ, in this case the intestine, results in improved feed, as well as having a lower environmental impact. However, the degradation of bioactive molecules' in transit in the gastrointestinal passage is still an unresolved issue. This paper discusses the feasibility of a simple and cost-effective procedure to bypass the degradation problem. A solid/liquid adsorption procedure was applied, and the operating parameters (pH, reaction time, and LY initial concentration) were studied. Lysozyme is used in this work as a representative bioactive molecule, while Adsorbo®, a commercial mixture of clay minerals and zeolites which meets current feed regulations, is used as the carrier. A maximum LY loading of 32 mgLY/gAD (LY(32)-AD) was obtained, with fixing pH in the range 7.5-8, initial LY content at 37.5 mgLY/gAD, and reaction time at 30 min. A full characterisation of the hybrid organoclay highlighted that LY molecules were homogeneously spread on the carrier's surface, where the LY-carrier interaction was mainly due to charge interaction. Preliminary release tests performed on the LY(32)-AD synthesised sample showed a higher releasing capacity, raising the pH from 3 to 7. In addition, a preliminary Trolox equivalent antioxidant capacity (TEAC) assay showed an antioxidant capacity for the LY of 1.47 ± 0.18 µmol TroloxEq/g with an inhibition percentage of 33.20 ± 3.94%.