Capacity Assay Research Articles

Evaluation of Biological Activity of Essential Oil from Teucrium polium L.

Background: Teucrium polium L. has an essential place in traditional medicine throughout the world. Their use, based on empirical knowledge passed down from generation to generation, has helped to treat a multitude of illnesses and relieve a variety of symptoms. These traditional practices have laid the foundations for modern research, which seeks to understand the significant antibacterial properties. The bioactive compounds present in the plant, such as essential oils and polyphenols, have demonstrated antimicrobial activity against a wide range of pathogenic bacteria. This includes bacteria responsible for gastrointestinal, skin and respiratory infections. Methods: The focus of this study was the extraction of essential oils from the aerial portions of Teucrium polium L. via the hydrodistillation method, their anti-oxidant effect using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging assay method and iron reduction and antimicrobial activity of these essential oils was evaluated against seven ATCC bacterial strains: Staphylococcus aureus ATCC25923, Escherichia coli ATCC25922, Salmonella typhimurium ATCC10428, Pseudomonas aeruginosa ATCC27853, Enterococcus faecalis ATCC29212, Klebsiella pneumoniae ATCC700603 and Bacillus subtilis ATCC7033. In addition, the minimum inhibitory concentrations (MICs) were determined by incorporation method. These tests were carried out at the Laboratoires de the analytical chemistry laboratory at the Department of Pharmacy, Badji Mokhtar University, Annaba, Algeria. Result: The hydro distillation method yielded an essential oil of 0.45%. Antimicrobial activity was tested by the sensitivity test, which revealed inhibition zone diameters ranging from 11.5 mm to £20 mm. In addition, minimum inhibitory concentrations (MICs) were determined, showing variations between bacterial strains, suggesting potential therapeutic importance for some strains. Additionally, the antioxidant activity of the essential oils was assessed using the iron-reducing capacity assay and the DPPH free radical scavenging assay. Although the results indicated notable antioxidant potential, the IC50 values obtained were relatively high when compared to ascorbic acid, registering 107.38 mg/ml for iron reduction and 91.62 mg/ml for DPPH reduction. These findings contribute to the exploration of the therapeutic potential of Teucrium polium L. essential oils for their antimicrobial and antioxidant properties.

A Paper-Based Assay for the Determination of Total Antioxidant Capacity in Human Serum Samples.

Determining the total antioxidant capacity (TAC) of biological samples is a valuable approach to measuring health status under oxidative stress conditions, such as infertility and type 2 diabetes. The Trolox equivalent antioxidant capacity (TEAC) assay is the most common approach to evaluating TAC in biological matrices. This assay is typically performed in clinical settings on a microtiter plate using a plate reader. However, the instrumentation and expertise requirements, and the resulting delay in the reporting of assay outcomes, make solution-based TEAC assays impractical for point-of-care or at-home testing, where individuals may want to monitor their health status during treatment. This work introduces the first microfluidic paper-based analytical device (µPAD) that measures TAC in human serum using TEAC assay chemistry. TAC was determined through a colorimetric image analysis of the degree of decolorization of 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) radical cations (ABTS●+) by serum antioxidants. The µPAD showed a linear response to Trolox, ranging from 0.44 to 2.4 mM, (r = 0.999). The performance of paper-based TEAC assays was validated through direct comparison to solution-based TEAC assays. There was a 0.04 mM difference in TAC values between the two platforms, well within one standard deviation of a standard solution-based assay conducted on an aliquot of the same serum sample (±0.25 mM). The µPAD had a limit of detection (LOD) of 0.20 mM, well below the TAC of normal human serum. The results suggest that the proposed device can be used for biological TAC determination and expands the field of TAC analysis in point-of-care health monitoring.

Total Antioxidant Status in Critically Ill Patients with Traumatic Brain Injury and Secondary Organ Failure-A Systematic Review.

Introduction: The available literature indicates that oxidant-antioxidant imbalance plays a significant role in the pathophysiology of traumatic brain injury and the subsequent secondary organ dysfunctions. However, there is a lack of studies summarizing the knowledge in this area, and no clear guidelines exist regarding the use of biomarkers of oxidative stress as diagnostics tools. Methods: The present work aims to provide a systematic review of the literature on the use of total antioxidant capacity (TAC) assays in predicting the outcomes of traumatic brain injury (TBI). A literature search was conducted up to 1 September 2024, according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA 2020) guidelines, using the PubMed and Scopus databases. Based on the inclusion criteria, 24 studies were used for the final review. Results: Promising data indicate that TAC assays are useful in predicting 30-day mortality and neurological outcomes. Moreover, they correlate with radiological findings on CT scans in brain injury and the clinical classifications of injuries, as well as the parameters of organ failure. Conclusions: Total antioxidant capacity assays can be used to assess the extent of brain damage and prognosticate general vital functions. Future experiments should include long-term randomized clinical trials on larger populations of TBI patients.

Discovery and synthesis of novel phenoxyacetate ester Schiff base α-glucosidase inhibitors

A phenoxyacetate ester Schiff base lead compound 4a (ZINC20073984) was firstly discovered through molecular docking screening and molecular dynamics (MD) simulation, which could be used as an α-glucosidase (PDB: 3A4A) inhibitor. Then a series of α-glucosidase inhibitors 4b-4r with novel structures were designed and synthesized with the lead ZINC20073984 as a template. The results of in vitro α-glucosidase inhibitory activity show that the synthesized phenoxyacetate ester Schiff base compounds 4e, 4o-4r (IC50 values range from 5.44 ± 0.52 μM to 33.67 ± 9.1 μM) exhibit good activity. Among them, ethyl (Z)-2-(2,4-dinitro-5-(2-(1-(2,4,6-trimethoxyphenyl) ethylidene) hydrazineyl) phenoxy) acetate (4r) and ethyl (Z)-2-(2,4-dinitro-5-(2-(1-(1-(4-ethoxyphenyl) ethylidene) hydrazineyl) phenoxy) acetate (4p) exert the best inhibitory activity, with IC50 values of 5.44 ± 0.52 μM and 5.80 ± 1.4 μM, respectively, which are superior to the standard drug acarbose (IC50 = 8.36 ± 0.02 μM). Molecular docking results indicate that the good inhibitory activity of 4r and 4p may be attributed to multiple hydrogen-bonding interactions with the α-glucosidase. Furthermore, the drug-likeness of all synthesized compounds was evaluated using the pkCSM tool, and ADMET predictions were conducted for compound 4r. The results demonstrated that all compounds follow Lipinski’s rules, and 4r possesses favorable pharmaceutical properties. In an in vitro cytotoxicity assay, the most potent 4r shows non-cytotoxicity to the 3T3 cells with an CC50 value > 40 µM. In both antioxidant and anti-cancer capacity assays, compound 4r has demonstrated promising potential. The results of this study might be helpful in the discovery of new drugs for the treatment of diabetes mellitus type-2 and bladder cancer.

Antioxidant Potential Evaluation at Various Stages of Black Cumin Oil Production.

Nigella sativa L. seeds and their industrial process products, oils, cake, and meal, are valuable sources of bioactive compounds with antioxidant properties. In this work, the effect of technological processes on the antioxidant capacity (AC) and total phenolic content (TPC) in the black cumin oils obtained by cold pressing and solvent extraction, as well as the by-products, were evaluated. The AC values of black cumin seeds (BCS), cold-pressed black cumin oil (BCCPO), black cumin oil extracted from seeds (BCEO-S), black cumin oil extracted from cake (BCEO-C), black cumin cake (BCC), and black cumin meal (BCM) were determined by 2,2-diphenyl-1-picrylhydrazyl (DPPH) and cupric reducing antioxidant capacity (CUPRAC) assays, whereas TPC in these samples was analyzed by the Folin-Ciocalteu (FC) method. Two applied conventional oil extraction methods, screw pressing and solvent extraction, significantly affected the AC and TPC in the obtained black cumin oils and by-products. The solvent-extracted black cumin oils revealed higher antioxidant properties (DPPH = 4041-16,500 μmol TE/100 g, CUPRAC = 1275-4827 μmol TE/100 g) than the cold-pressed black cumin oil (DPPH = 3451 μmol TE/100 g and CUPRAC = 3475 μmol TE/100 g). In addition, the oil yield (20.92-48.86%) and antioxidant properties of BCCPO (DPPH = 2933-5894 μmol TE/100 g and TPC = 135-199 mg GAE/100 g) and BCC (DPPH = 1890-2265 μmol TE/100 g and TPC = 284-341 mg GAE/100 g) closely depended on the nozzle diameters (5, 8, and 10 mm) mounted in a screw press. Although both by-products were a rich source of antioxidants, BCM had significantly lower CUPRAC (1514 μmol TE/100 g) and TPC (92 mg GAE/100 g) values than BCC (CUPRAC = 3397 μmol TE/100 g and TPC = 426 mg GAE/100 g). Nevertheless, acid hydrolysis and alkaline hydrolysis of BCM extracts significantly increased their antioxidant potential. However, the DPPH (35,629 μmol TE/100 g), CUPRAC (12,601 μmol TE/100 g), and TPC (691 mg GAE/100 g) results were higher for the BCM extract after acid hydrolysis than those for alkaline hydrolysate (DPPH = 2539 μmol TE/100 g, CUPRAC = 5959 μmol TE/100 g, and TPC = 613 mg GAE/100 g). Finally, the generated AGREEprep metrics highlighted the sustainability and the greenness of the cold pressing of oil from BCS.

Measuring Oxidative Stress by the Iridium Reducing Capacity Assay

Measuring Oxidative Stress by the Iridium Reducing Capacity Assay

Evaluation of Total Flavonoids, Total Phenolics Content and in Vitro Antioxidant Activities of Extract of Platycerium bifurcatum Leaf: A Potential Antihypertensive Medicinal Plant

Plant bioactive compounds possess the potential to function as antioxidants, which makes interest in the use of ethnomedicine for treatment or prevention of diseases associated with oxidative stress to be growing. This study evaluated the flavonoid contents, total phenolics and in vitro antioxidant activity of Platycerium bifurcatum leaf extract. The free radical scavenging activity was determined using diphenyl-picrylhydrazyl (DPPH) and nitric oxide assay, and the reducing power by ferric reducing antioxidant power (FRAP) and the total antioxidant capacity assays. The total phenolics content of the extract was 4.7 ± 0.56 mg GAE/g extract and flavonoid contents of the extract was 46.73 ± 2.03 mg QE/g extract. The extarct exhibited a dose dependent nitric oxide and DPPH free radical scavenging capacity, which showed a strong correlation between the antioxidant activity and the flavonoid contents and total phenolics. The appreciable amount of flavonoid contents, total phenolics and its strong antioxidant properties suggested that Platycerium bifurcatum is a promising source of novel lead antioxidants and could be utilized in the treatment of oxidative stress mediated cardiovascular disease like hypertensive and obesity

Phytochemical characterization and valorization of Quercus ilex (west of Algeria)

<p><span style="font-family:"Times New Roman","serif";font-size:12pt;line-height:115%;"><strong>ABSTRACT</strong></span></p><p style="text-align:justify;"><span style="font-family:"Times New Roman","serif";font-size:12pt;line-height:115%;" lang="EN-US">This project is part of the promotion of natural resources such as Non-Timber Forest Products (NTFP). In order to assess the economic value of NTFP, this research focuses on the phytochemical analysis of holm oak (</span><em><span style="font-family:"Times New Roman","serif";font-size:12pt;line-height:115%;" lang="EN-US">Quercus ilex </span></em><span style="font-family:"Times New Roman","serif";font-size:12pt;line-height:115%;" lang="EN-US">L.) from the Terni region (Western Algeria). It focused on bioactive substances, particularly polyphenols. The method used consists of extracting the phenolic compounds by two methods which are maceration and ultrasound from the leaves and bark of this species. The quantity of these compounds is assessed spectrometrically and the antioxidant activity of the extracts in vitro using free radical scavenging and total antioxidant capacity assays. The results obtained indicate that maceration offers the best yield for extracting phenolic compounds from the leaves, with a yield of 10.74%. These results also showed that the extracts of holm oak leaves and bark are rich in phenolic compounds and have a content of total phenols (103,014±1,393 and 813.462±1.675 mg EAG/g DM), flavonoids (87.078±6.220 and 949.936±36.394 mg EC/g DM) and condensed tannins (57.194±12.447 and 940.195±20.417 mg EC/g DM). Regarding the antioxidant power, it is evident that all extracts have significant antioxidant capacity, and all have high free radical scavenging activity. The highest inhibition percents are 84.98±7.45% for the leaf extract and 83.15±9.27% for the bark extract. According to these results, holm oak contains a large quantity of polyphenols and antioxidant molecules. This information represents a natural source of antioxidants which remains to be exploited for future use in the agri-food and health sectors, within the broader framework of promoting the biodiversity of Algerian plants (NTFP).</span></p>

Characteristics of Polysaccharides from Industrial Hemp (Cannabis sativa L.) Kernels.

Polysaccharides from hemp seeds exhibit antioxidant activities in vitro and in vivo. However, crude polysaccharide quality is often low owing to the presence of fibres and pigment impurities, which are difficult to eliminate in the hemp seed shell. In this study, crude polysaccharides from hemp kernels (HKP) were obtained by hot water extraction and separated by membrane ultrafiltration into eight fractions with different molecular weights. Total antioxidant capacity and free radical scavenging (DPPH) assays were performed to evaluate the antioxidant activities of HKP and the fractions in vitro. The structural characteristics of HKP were determined using various analytical techniques. The Fe3+-reducing power of HKP was 7.65 ± 0.22 μmol/g, and HKP possessed the highest DPPH radical-scavenging rates (94.30 ± 2.27%), similar to 5 mg/mL Vitamin C (Vc), which had a rate of 95%. The HKP was an acidic polysaccharide with a low molecular weight (4.21 ± 0.12 kDa). The monosaccharide composition indicated that HKP primarily comprised mannose, ribose, glucuronic acid, galacturonic acid, glucose, galactose, arabinose, and fucose in a molar ratio of 0.96:1.95:8.27:0.98:9.46:1.69:6.10:2.82. The molar mass of HKP was distributed widely in a triple helical conformation. This study provides a scientific basis for further research on the use of hemp polysaccharides.

Characterization and monitoring of changes during lactation in the profile of multiple bioactive compounds of milk from grazing mares.

Mare milk has often been considered a food product with potential functional properties. However, the bioactive compound composition of mare milk, including vitamins and other minor bioactive compounds, as well as factors affecting this composition have scarcely been studied. Therefore, the present study aimed to characterize the changes during lactation in the content of water- and fat-soluble vitamins and total polyphenols, and the total antioxidant capacity of mare milk from semi-extensive farms. A total of 310 individual milk samples from 18 mares belonging to three commercial farms and 12 lactation times were analyzed. Ascorbic acid (vitamin C), thiamine (vitamin B1), riboflavin (vitamin B2), nicotinic acid and niacinamide (vitamins B3), pantothenic acid (vitamin B5), pyridoxal and pyridoxine (vitamins B6), folic acid (vitamin B9), cyanocobalamin (vitamin B12), tocopherols and tocotrienols (vitamin E) and retinol and retinyl esters (vitamin A) were quantified using liquid chromatography. Total polyphenols and antioxidant capacity assays were analyzed using spectrophotometry. The concentration of most bioactive compounds tended to decline as lactation progressed, with the exception of polyphenols and the total antioxidant capacity that oscillated during lactation. On the other hand, the effect of the different semi-extensive management of the farms was only significant for vitamin B3 content. To the best of our knowledge, the present study provides the most in-depth description of the vitamin profile of mare milk as well as new insights into polyphenol content and antioxidant capacity of mare milk. © 2024 The Author(s). Journal of the Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Ultrathin PtTeNi nanosheets as a peroxidase mimic for colorimetric detection of ascorbic acid

Utilizing the peroxidase (POD) activity of nanozymes to construct the colorimetric sensors is a popular strategy for assessing the antioxidant capacity of biological molecules, but the detection sensitivity is often compromised by their low enzymatic activity. Pt-based nanozymes, favored by their regulated nanostructures, well provide numerous binding sites for H2O2, and exhibit enhanced POD performance. Here, we utilized a simple hydrothermal method to synthesize PtTeNi nanomaterials featured with the ultrathin sheets for detecting biological antioxidant (ascorbic acid (AA) as the model). With the TMB-H2O2 system as the colorimetric substrate, PtTeNi nanomaterial exhibited superior POD-like activity with a higher Vmax (5.02 × 10−4 mM s−1) and lower Km (3.14 mM)) in contrast to natural HRP. AA significantly inhibited the colorimetric reaction of PtTeNi-TMB-H2O2 system via competitively binding OH generated by PtTeNi-mediated H2O2 catalysis and further blocking oxTMB formation. Inspired by this fact, the PtTeNi-TMB-H2O2 colorimetric sensor successfully realized highly sensitive and selective detection of AA with a good linear range from 5 μM to 500 μM and the detection limit of 2.94 μM. For analyzing the total antioxidant capability and AA contents in actual samples (commercial drinks and vitamin C tablets), the PtTeNi-based colormetric method displayed comparable results with the commercial total antioxidant capacity assay kit, with the advantages of easy operation, convenient storage and high stability. This work fully affirms the excellent enzyme-like performance of Pt-based nanomaterial and confirms the feasibility of nanozymes-based colorimetric sensing strategy in detecting antioxidant capability of biological molecules.

Characterization, biological activity, and anticancer effect of green-synthesized gold nanoparticles using Nasturtium officinale L.

BackgroundNanostructured materials used have unique properties and many uses in nanotechnology. The most striking of these is using herbal compounds for the green synthesis of nanoparticles. Among the nanoparticle types used for green synthesis, gold nanoparticles (AuNPs) are used for cancer therapy due to their stable structure and non-cytotoxic. Lung cancer is the most common and most dangerous cancer worldwide in terms of survival and prognosis. In this study, Nasturtium officinale (L.) extract (NO), which contains biomolecules with antioxidant and anticancer effects, was used to biosynthesize AuNPs, and after their characterization, the effect of the green-synthesized AuNPs against lung cancer was evaluated in vitro.MethodsUltraviolet‒visible (UV‒Vis) spectrophotometry, scanning electron microscopy (SEM), transmission electron microscopy (TEM), energy dispersive X-ray spectroscopy (EDS), multiple analysis platform (MAP), and Fourier transform infrared (FT-IR) spectroscopy analyses were performed to characterize the AuNPs prepared from the N. officinale plant extract. Moreover, the antioxidant activity, total phenolic and flavonoid contents and DNA interactions were examined. Additionally, A549 lung cancer cells were treated with 2–48 µg/mL Nasturtium officinale gold nanoparticles (NOAuNPs) for 24 and 48 h to determine the effects on cell viability. The toxicity of the synthesized NOAuNPs to lung cancer cells was determined by the 3-(4,5-dimethylthiazol-2-il)-2,5-diphenyltetrazolium bromide (MTT) assay, and the anticancer effect of the NOAuNPs was evaluated by apoptosis and cell cycle analyses using flow cytometry.ResultsThe average size of the NPs was 56.4 nm. The intensities of the Au peaks from EDS analysis indicated that the AuNPs were synthesized successfully. Moreover, the in vitro antioxidant activities of the NO and NOAuNPs were evaluated; these materials gave values of 31.78 ± 1.71% and 31.62 ± 0.46%, respectively, in the 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay at 200 g/mL and values of 25.89 ± 1.90% and 33.81 ± 0.62%, respectively, in the 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) assay. The NO and NOAuNPs gave values of 0.389 ± 0.027 and 0.308 ± 0.005, respectively, in the ferrous ion reducing antioxidant capacity (FRAP) assay and values of 0.078 ± 0.009 and 0.172 ± 0.027, respectively, in the copper ion reducing antioxidant capacity (CUPRAC) assay. When the DNA cleavage activities of NO and the NOAuNPs were evaluated via hydrolysis, both samples cleaved DNA starting at a concentration of 25 g/mL in the cell culture analysis, while the nanoformulation of the NO components gave greater therapeutic and anticancer effects. We determined that the Au nanoparticles were not toxic to A549 cells. Moreover, after treatment with the half-maximal inhibitory concentration (IC50), determined by the MTT assay with A549 cells, we found that at 24 and 48 h, while the necrosis rates were high in cells treated with NO, the rates of apoptosis were greater in cells treated with NOAuNPs. Notably, for anticancer treatment, activating apoptotic pathways that do not cause inflammation is preferred. We believe that these results will pave the way for the use of NOAuNPs in in vitro studies of other types of cancer.ConclusionIn this study, AuNPs were successfully synthesized from N. officinale extract. The biosynthesized AuNPs exhibited toxicity to and apoptotic effects on A549 lung cancer cells. Based on these findings, we suggest that green-synthesized AuNPs are promising new therapeutic agents for lung cancer treatment. However, since this was an in vitro study, further research should be performed in in vivo lung cancer models to support our findings and to explain the mechanism of action at the molecular level.

The combination of allantoin, bisabolol, D-panthenol and dipotassium glycyrrhizinate mitigates UVB-induced PGE2 synthesis by keratinocytes.

Erythema, characterized by the redness of the skin, is a common skin reaction triggered by various endogenous and exogenous factors. This response is often a result of the activation of underlying inflammatory mechanisms within the skin. The objective of this study is to investigate the potential benefits of applying a combination of skincare ingredients, namely allantoin, bisabolol, D-panthenol and dipotassium glycyrrhizinate (AB5D), in the modulation of inflammatory factors associated with erythema. Additionally, the study aims to elucidate the mechanisms by which these ingredients exert their combined actions to alleviate erythema-associated inflammation. Human epidermal keratinocytes were exposed to UVB and subsequently treated with AB5D. Transcriptomics profiling was performed to analyse the dose-response effect of AB5D treatment on keratinocytes. The quantitation of inflammatory mediators, including PGE2, IL-1α, IL-6, IL-8, IL-1RA and TNFα, was performed on cultured media. Additionally, the oxygen radical absorbance capacity (ORAC) assay was carried out to evaluate the total antioxidant capacity of both individual ingredients and the AB5D combination. To assess the in-vitro antioxidant effects of AB5D against UVB-induced oxidative stress in hTERT keratinocytes, real-time quantitation of mitochondrial superoxide was measured through live-cell imaging. The application of AB5D to UVB-exposed keratinocytes downregulated gene sets associated with inflammatory responses, highlighting the anti-inflammatory properties of AB5D. Specifically, AB5D effectively reduced the production of PGE2, leading to the downregulation of inflammatory cytokines. Moreover, our findings indicate that AB5D exhibits antioxidative capabilities, functioning as both an antioxidant agent and a regulator of antioxidant enzyme expression to counteract the detrimental effects of cellular oxidative stress. We demonstrated that AB5D can reduce UVB-induced PGE2, IL-1α, IL-6, IL-8, IL-1RA and TNFα as well as mitochondrial superoxide. These findings suggest that AB5D may alleviate erythema by modulating inflammation via PGE2 and through antioxidation mechanisms.

Phytochemical Screening, Antioxidant, Cytotoxic, Analgesic, Antidiarrheal Activity and GC-MS Analysis of the Leaves of Stachyphrynium Placentarium

The goal of the present study is to analyze the phytochemicals, antioxidants, cytotoxic, analgesic and anti-diarrheal properties of the leaves of Stachyphrynium placentarium methanol extract as well as its dichloromethane soluble fraction using GC-MS technique. The leaves of the S. placentarium were extracted with methanol and designated as SPM. Phytochemicals investigation exhibited the presence of several important secondary metabolites. Antioxidants such as total phenolic content, flavonoids content, total antioxidant, reducing power capacity and DPPH (2,2-diphenyl-1-picrylhydrazyl) free radical scavenging assay SPM revealed non-significant antioxidant properties. The cytotoxicity study of the extract showed significant cytotoxicity (LC50 35.1093 µg/mL) compared to standard vincristine sulfate (LC50 0.482 µg/mL). Analgesic study of S. placentarium by hot plate model indicates both the doses exhibited significant central analgesic activity compared with the control. The maximum analgesic activity was observed at 90 min. SPM (500 mg/kg) is 60.21 %. SPM250 also showed a significant percentage of elongation or latency from 30 to 150 min 57.00, 59.64, 59.88, 58.07 and 53.71 %, respectively 30 min time interval. In the acetic acid writhing test, S. placentarium significantly outperformed the control and standard (SPM250 - 52.27 %, SPM500 - 34.09 and STD - 27.27 %), showing a significant increase in peripheral analgesic activity (p < 0.001). Antidiarrheal activity of SPM showed that SPM250 and SPM500 substantially decreased the feces in castor oil-induced diarrhea (p < 0.05). According to the study, the incidence and severity of diarrhea are significantly reduced by SPM500. Four chemicals are found in the dichloromethane-soluble portion of the methanol extract of S. placentarium leaves, according to GC-MS studies such as Octadecane, 6-methyl- (1.326 %), Dodecane (1.02 %), 1-Iodo-2-methylundecane (0.47 %), tetradecanoic acid 12-methyl, methyl ester (72.64 %). Tetradecanoic acid exhibits anti-inflammatory, anticancer, 5-reductase inhibitory, antioxidant, hemolytic and antifibrinolytic properties. HIGHLIGHTS The phytochemicals, antioxidants, cytotoxic, analgesic and anti-diarrheal properties of Stachyphrynium placentarium methanol extract (SPM) leaves. The dichloromethane soluble fraction is examined using Gas chromatography–mass spectrometry (GC-MS) and identified Octadecane, 6-methyl-, Dodecane, 1-Iodo-2-methylundecane, tetradecanoic acid 12-methyl, methyl ester. The brine shrimp lethality test exhibited that SPM has significant cytotoxicity (LC50 35.1093 µg/mL) compared to vincristine sulphate (LC50 0.482µg/mL), the extract showed significant cytotoxicity (LC50 35.1093 µg/mL). S. placentarium at both dosages (250 and 500 mg) provided significant central analgesia compared to the control in a hot plate model. In the acetic acid writhing test, S. placentarium showed significantly higher peripheral analgesic action (SPM250 - 52.27, SPM500 - 34.09 and STD - 27.27 %) compared to control and standard. In castor oil-induced diarrhoea, SPM250 and SPM500 (250 and 500 mg) showed significantly reduced faeces production (p < 0.05). GRAPHICAL ABSTRACT

Phytochemical Analysis and Evaluation of Antioxidant, Antidiabetic, and Anti-inflammatory Properties of Aegle marmelos and Its Validation in an In-Vitro Cell Model.

Persistent hyperglycemia significantly increases oxidative stress and inflammation resulting in multiple cellular and molecular alterations which further exacerbate the diabetes associated complications.Aegle marmelos (L.) Corrêais a medicinal plant used in the Indian system of medicine for treating various disorders including diabetes. However, studies on phytoconstituents and their pharmacological activity of this plant are limited. Therefore, we aimed to determine the phytochemical components, evaluate the antidiabetic activity, anti-inflammatory activity, and antioxidant activity ofA. marmelosleaf extract, and validate its mechanistic effects in an in vitro cell model. The qualitative and quantitative analysis of the different phytoconstituents in the extract was determined using standardized protocols. The antioxidant activity of the extract was evaluated by 2,2-di-phenyl-1-picrylhydrazyl (DPPH) radical scavenging capacity assay and ferric reducing antioxidant power (FRAP) assay. The antidiabetic activity of the extract was evaluated by α-amylase inhibition and α-glucosidase inhibition assay. The anti-inflammatory activity was studied using an albumin denaturation assay. In addition, the pharmacological effect(s) of leaf extract was checked in the normal rat kidney fibroblast cells (NRK-49F) under high glucose conditions. Intracellular reactive oxygen species (ROS) generation was measured by fluorometry using fluorescence probe 2',7'-dichlorodihydrofluorescin diacetate (DCF-DA). mRNA expression of inflammatory markers including inducible nitric oxide synthase (iNOS) and tumor necrosis factor-alpha (TNF-α) was studied using real-time quantitative polymerase chain reaction (RT-qPCR). Cell migration was studied using cell scratch assay. Statistical analysis was performed using GraphPad Prism version 8.0. The phytochemical analysis ofA. marmelosleaf extract revealed the presence of alkaloids, phenols, flavonoids, and saponins. The extract showed higher antioxidant activity in the DPPH (IC50=258.21 µg/mL) and FRAP assay (IC50=293.83 µg/mL). The extract exhibited prominent antidiabetic activity by inhibiting enzymes α-Amylase (IC50=73.2 µg/mL) and α-glucosidase (IC50=43.9 µg/mL). In addition, the extract showed effective anti-inflammatory activity by significantly inhibiting the denaturation of egg albumin (IC50=102.8 µg/mL). Further, the leaf extract significantly decreased the high glucose-induced ROS generation as well as inflammatory markers in rat fibroblast cell lines in a dose-dependent manner. Additionally, high glucose-induced cell migration as the measure of cell injury was effectively reduced by the extract treatment. A. marmelosleafextract was quantified to possess a substantial amount of important phytoconstituents that have promising pharmacological properties. Besides showing antidiabetic activity, the extract significantly combats the high glucose-induced ROS generation, inflammatory markers expressions, and cell migration. Further, in-depth studies and clinical trials are warranted so as to position these traditional remedies for the treatment of metabolic disorders.

Colchicine, serotobenine, and kinobeon A: novel therapeutic compounds in Carthamus tinctorius L. for the management of diabetes

Diabetes, a global health concern, poses increasing mortality risks. The pathogenesis of diabetes involves multiple mechanisms, with oxidative stress being one of the key contributors. As synthetic drugs have various side effects, which can be minimized by using herbal plants. This study focuses on the In vitro antioxidant potential, α-amylase inhibition potential, identification of bioactive compounds, and hub genes in diabetes treatment mechanism by using C. tinctorius Extraction of C. tinctorious lead and flower was performed using different solvents (Distilled water, methanol, chloroform, and Dimethyl ether). After extraction different concentrations range from 25–200 mg/mL) was made and checked against activities. The antioxidant potential was assessed using 2, 2-diphenyl-1-picrylhydrazyl (DPPH), total phenolic contents (TPC), and total antioxidant capacity (TAC) assays, while antidiabetic activity was evaluated through α-amylase inhibition assay. Phytochemicals was identified by GC–MS analysis, followed by ADMET screening and network pharmacology analysis using Swiss Target Prediction, Gene Card, DesGeNet, DAVID, STRING, Cytoscape, and drug revitalization databases. Results revealed positive correlations with DPPH, TAC, and TPC. Methanol extract exhibited the highest inhibitory concentration. Screening of 46 compounds was performed by studying their pharmacokinetic properties which revealed 9 compounds effective against 204 diabetes targets. Moreover, their network analysis identified four hub genes, including AKT1, JUN, EGFR, and MMP9. These genes found highly associated with drugs like Colchicine and Serotobenine. Revitalization analysis also highlighted four genes (EGFR, PTGS2, AKT1, and MMP9) strongly correlated with FDA-approved drugs. The study suggests C. tinctorius methanol extract is a potential source for novel drugs.Graphical

Chemical profile and biological activities of Lysiphyllum binatum (Blanco) de Wit.

Lysiphyllum binatum (Blanco) de Wit in the Fabaceae family, despite its traditional medicinal uses, has not been the subject of prior scientific inquiry into its chemical and biological profile. The dichloromethane and MeOH extracts of its roots exhibited notably similar antioxidant activity, while the dichloromethane extract of the vine stems showed aromatase inhibition. This study aimed to identify the bioactive components responsible for these activities. Chemical investigation of the roots led to the isolation of six new metabolites, named lysiphans A-F (1-6), along with eight known compounds (7-14). The vine stem yielded lysiphan C (3) and compound 7, as well as five known isolates (15-19). The structures of these metabolites were determined through NMR spectral analysis, HRESIMS, quantum chemical calculations of NMR and ECD spectra, and Mosher's modifications to establish their absolute configurations. The biogenetic relationships between the new compounds were proposed. Several of the isolates were evaluated for their antioxidant, anti-aromatase, and cytotoxic properties. Lysiphan B (2) exhibited significant antioxidant activity, with an IC50 value of 28.8 ± 0.4μM in the diphenyl picrylhydrazyl radical (DPPH) assay, 3.5 ± 0.2μM in the xanthine/xanthine oxidase (XXO) assay, and 1.5 ± 0.0 ORAC units in oxygen radical absorbance capacity (ORAC) assay. Additionally, compounds 12, 13, and 16 exhibited very strong aromatase inhibitory activity with IC50 values of 0.3 ± 0.2, 4.7 ± 0.1, and 0.9 ± 0.2µM, respectively. Compound 16 also demonstrated strong ORAC activity of 1.9 ± 0.1 ORAC units.

Evaluation of the bioactivity of Berberis microphylla G. Forst (Calafate) leaves infusion

Berberis microphylla G Forst (Calafate) have been used in traditional medicine from prehispanic times in Patagonia. In the last decade the consumption of the fruit has been increased due to their antioxidant capacity, and because several studies demonstrated health benefits associated with the protection against atherosclerosis and other metabolic diseases. Nevertheless, the bioactivity properties of the leaves, a by-product of agronomic management, have been poorly studied. Recently, 108 compounds mainly hydroxycinnamic acids, flavonols, and berberine were identified in a methanolic extract of the leaves, demonstrating great potential for the development of new functional beverages. Based on these, for first time a comprehensive chemical characterization and bioactivity was evaluated for a Calafate leaves infusion prepared in hot water. For this, chemical characterization of the infusion was performed by UHPLC-Q-TOF and TXRF. Bioactivity was assayed by antioxidant capacity, cell cytotoxicity, and cell oxidative stress assays. Inhibition of both Aβ aggregation for Alzheimer's disease and gastrointestinal enzymes for metabolic syndromes were evaluated. The results show that the infusion is rich in hydroxycinnamic acids and other bioactive compounds. The infusion does not contain toxic metals or cytotoxicity activity. The infusion can reduce intracellular reactive oxygen species in HUVEC cells and showed a reduction in the Aβ aggregation being a potential beverage for Alzheimer's prevention. Finally, the infusion had in-vitro hypoglycemic and hypolipidemic effects. These results support the usage of Berberis microphylla G Forst leaves as a new functional beverage.

Exploring antioxidative properties of xanthohumol and isoxanthohumol: An integrated experimental and computational approach with isoxanthohumol pKa determination

This study explores the antioxidative activities of xanthohumol (XN) and isoxanthohumol (IXN), prenylated flavonoids from Humulus lupulus (family Cannabaceae), utilizing the oxygen radical absorption capacity (ORAC) and ferric reducing antioxidant power (FRAP) assays along with computational Density Functional Theory methods. Experimentally, XN demonstrated significantly higher antioxidative capacities than IXN. Moreover, we determined IXN pKa values using the UV/Vis spectrophotometric method for the first time, facilitating its accurate computational modeling under physiological conditions. Through a thermodynamic approach, XN was found to efficiently scavenge HOO• and CH3O• radicals via Hydrogen Atom Transfer (HAT) and Radical Adduct Formation (RAF) mechanisms, while CH3OO• scavenging was feasible only through the HAT pathway. IXN exhibited its best antioxidative activity against CH3O• via both HAT and RAF mechanisms and could also scavenge HOO• through RAF. Both Single Electron Transfer (SET) and Sequential Proton Loss-Electron Transfer (SPLET) mechanisms were thermodynamically unfavorable for all radicals and both compounds.

Synthesis, electrochemical properties, and antioxidant activity of sterically hindered catechols with 1,3,4-oxadiazole, 1,2,4-triazole, thiazole or pyridine fragments.

A series of new RS-, RS-CH2- and R2N-CH2-functionalized сatechols with heterocyclic fragments such as 1,3,4-oxadiazole, 1,2,4-triazole, thiazole, or pyridine were synthesized by the reaction of 3,5-di-tert-butyl-o-benzoquinone or 3,5-di-tert-butyl-6-methoxymethylcatechol with different heterocyclic thiols. The S-functionalized catechols were prepared by the Michael reaction from 3,5-di-tert-butyl-o-benzoquinone and the corresponding thiols. The starting reagents such as substituted 1,3,4-oxadiazole-2-thiols and 4H-triazole-3-thiols are characterized by thiol-thione tautomerism, therefore their reactions with 3,5-di-tert-butyl-6-methoxymethylcatechol can proceed at the sulfur or nitrogen atom. In the case of mercapto-derivatives of thiazole or pyridine, this process leads to the formation of the corresponding thioethers with a methylene linker. At the same time, thiolated 1,3,4-oxadiazole or 1,2,4-triazole undergo alkylation at the nitrogen atom in the reaction with 3,5-di-tert-butyl-6-methoxymethylcatechol to form the corresponding thiones. The yield of reaction products ranges from 42 to 80%. The crystal structures of catechols with 3-nitropyridine or 1,3,4-oxadiazole-2(3H)-thione moieties were established by single-crystal X-ray analysis. The possibility of forming intra- and intermolecular hydrogen bonds has been established for these compounds. The electrochemical behavior of the studied compounds is influenced by several factors: the nature of the heterocycle and its substituents, the presence of a sulfur atom in the catechol ring, or a thione group in the heterocyclic core. The radical scavenging activity and antioxidant properties were determined using the reaction with synthetic radicals, the cupric reducing antioxidant capacity assay, the inhibition process of superoxide radical anion formation by xanthine oxidase, and the process of lipid peroxidation of rat liver (Wistar) homogenates in vitro.