Protein Denaturation Assay Research Articles

Bioactive Phenolics and Flavonoids, Antioxidants, Anti-Inflammatory, Enzyme-Inhibitory and Cytotoxic Activities of Aerial part of Trachyspermum roxburghianum (DC.) Craib.

Trachyspermum roxburghianum (DC.) Craib is an edible plant found in Northern and Northeastern Thailand. This plant is used in household remedies to relieve carminative and digestive problems, hiccups, vomiting, bladder pain, etc. This study aims to assess the bioactive compounds, consisting of phenolics and flavonoids, and evaluate the biological activities of this plant. The aerial portion was macerated in EtOH1 to obtain EtOH1 crude extracts. Then, the EtOH1 was divided and separated by the solvents’ polarity to obtain Petr, EtOAc, and EtOH2 crude extracts. All crude extracts were assessed for their total phenolics and flavonoids and subsequentially evaluated for their antioxidants (using DPPH, ABTS and FRAP), enzymatic inhibitory effects (on tyrosinase and collagenase), anti-inflammatory activity (protein denaturation assay) and cytotoxicity (MTT assay). The results presented that a high amount of phenolics and flavonoids were observed in EtOH2 (56.57 mg GAE/g ext. and 22.75 mg QE/g ext.) followed by EtOAc (57.95 mg GAE/g ext. and 7.41 mg QE/g ext.), EtOH1 (47.14 mg GAE/g ext. and 4.23 mg QE/g ext.), and Petr (37.05 mg GAE/g ext. and 2.08 mg QE/g ext.), respectively. All crude extracts represented antioxidants in all assays; EtOAc presented a high level of antioxidants on ABTS (IC50 54.96 µg/mL) and DPPH (IC50 133.32 µg/mL), while EtOH2 revealed a high ability to reduce the Fe3+-TPTZ complex (155.85 Fe2+g ext.). Remarkably, for all crude extracts used for anti-collagenase enzyme activity, the percentage of inhibition ranged from 78.35 - 97.96 %. Besides, only Petr and EtOH2 showed the activity of the anti-protein denaturation (IC50 1.926 and 2.578 mg/mL). The cytotoxicity test showed that EtOH1, Petr, and EtOAc exhibited a concentration-dependent manner of inducing cell death in 24 and 48 h, while EtOH2 showed less toxicity to the cell line. This information would be valuable and pave the way for future study and application of this plant to other products. HIGHLIGHTS Trachyspermum roxburghianum (DC.) Craib from Northern Thailand High antioxidant and high anti-collagenase activities of Trachyspermum roxburghianum (DC.) Craib ethanolic extracts. GRAPHICAL ABSTRACT

GC-MS and HPTLC bioautography-based phytochemical profiling and evaluation of biological activity Neptunia prostrata Linn whole plant and leaves

This work used experimental methods to evaluate the chemical composition of GC-MS, HPTLC-based bioautographaphy, antioxidant, anti-diabetic, and protein denaturation properties of the Neptunia prostrata Linn (family: Mimosaceae) whole plant and leaves. Three methods, namely DPPH, metal chelating, and reducing power assays, were used to estimate the antioxidant effects. In addition, α-amylase, a carbohydrate-hydrolyzing enzyme, was targeted to test the anti-diabetic activity. The protein denaturation assay was used to evaluate the anti-inflammatory activity of the plant. GC-MS, an important analytical tool, was performed to identify bioactive compounds of medicinal plants used in pharmaceutical industries. Biochemical derivatization HPTLC-based bioautography with DPPH was executed to identify antioxidant agents from N. prostrata. The validated HPTLC method was used to quantify identified antioxidant agents. The stability of compounds, preparation of calibration curves, linearity, range, precision, limit of detection, limit of quantification, robustness, and accuracy are examined to develop a robust method. Higher total phenolic content (TPC) content was observed in N. prostrata hydroalcoholic extract of the whole plant (81.12 ± 4.19 mg GAE/g) and methanolic extract of leaves (105.16 ± 3.11 mg GAE/g). Methanolic extract of N. prostrata leaves showed significantly high antioxidant potential for all DPPH (IC50:28.05 ± 1.78 µg/mL), metal chelating (IC50:1598 ± 23.51 µg/mL), and reducing power assay compared to standard ascorbic acid (DPPH assay: IC50:24.14 ± 2.34 µg/mL and metal chelating: IC50:1301.16 ± 6.14 µg/mL). N. prostrata aqueous extract of the whole plant (IC50:179.47 ± 10.59 µg/mL) and leaves (IC50:130.86 ± 3.58 µg/mL) showed significantly high α-amylase inhibitory activity as compared to standard acarbose (IC50:148.62 ± 7.84 µg/mL). Methanolic extract of whole plants (IC50:131.46 ± 11.81 µg/mL) and chloroform extract of leaves (IC50:122.61 ± 8.19 µg/mL) exhibited strong anti-inflammatory activity as compared to standard acetylsalicylic acid (IC50:117.77 ± 4.95 µg/mL). GC-MS analysis revealed the presence of several bioactive compounds, where 2-cyclopenten-1-one, 2-hydroxy showed the most abundant bioactive compounds in the plant with 17.01%. HPTLC-bioautography identified three phenolic acids (chlorogenic acid, gallic acid, and 4-hydroxybenzoic acid) and one flavonoid (naringenin), further quantified with the validated HPTLC method. These findings propose that N. prostrata has therapeutic potential for treating diseases related to oxidative damage, diabetes, and inflammation, which may be due to the presence of bioactive compounds.

Integrating in silico molecular docking, ADMET analysis of C.verticillata with diabetic markers and in vitro anti-inflammatory activity

BackgroundOver the past decade, various research studies have proved the interconnection between the inflammatory pathways and diabetes complication in clinical condition. The present study evaluated the anti-inflammatory and antioxidant activity. Further, the sample was tested for its pharmacokinetics properties and the best compounds were docked with the diabetic markers (DPP IV (PDB-ID: IJ2E) and SGLT2 (PDB-ID: 7VSI)).ResultsC.verticillata showed a good hydrogen peroxide (78.3 ± 0.34%, IC50 = 287.81 µg/ml) and superoxide scavenging activity (52.7 ± 1.26%, IC50 = 796.15 µg/ml). In addition, the sample was checked for its anti-inflammatory activity with protein denaturation (57.4 ± 0.19%, IC50 = 471.5 µg/ml) and proteinase inhibition assay (68.3 ± 0.48%, IC50 = 213.42 µg/ml). Further, the bioactive compounds detected from HPLC-ESI-MS/MS analyzed sample were checked for its drug likeliness by checking its ADME properties and toxicological parameters. It has been observed that except Loliolide, all the other compounds have followed the physicochemical parameters and proved to exhibit drug likeliness characteristics. The bioactive compounds that follow the Lipinski’s rule were taken further for in silico molecular docking analysis with the diabetic protein markers (DPP IV and SGLT2). Docking results revealed that Pyro pheophorbide a with DPP IV and Dihydromonacolin L acid with SGLT2 have recorded a maximum docking score of (− 9.4 kcal/mol) and (− 9.2 kcal/mol), respectively.ConclusionThe observed results suggest that the identified and selected bioactive compounds from C.verticillata can be considered as a potential target molecule for the management of oxidative stress induced diabetic condition. Furthermore, the study also provides an insight on the effectiveness of the compounds on reducing the inflammation as well.

Anti-Inflammatory, Analgesic and Anti-Oxidant Effects of Shirakiopsis Indica (Willd). Fruit Extract: A Mangrove Species in the Field of Inflammation Research.

Shirakiopsis indica (Willd)., commonly known as Sa-Mor-Ta-Lay in Thailand, is a mangrove plant belonging to the Euphorbiaceae family. As mangrove plants' medicinal potentials are less explored, this study sought to qualitatively and quantitatively verify the bioactive components of Shirakiopsis indica fruits methanolic extract (SIF-ME) at the side of its analgesic, anti-inflammatory and anti-oxidant effects followed by in-silico studies. The in-vivo assessments of analgesic activity involved the hot plate test, acetic acid-induced writhing test, and formalin-induced licking test. The anti-inflammatory efficacy was assessed through the human RBC membrane stabilization assay (HRBC), protein denaturation assay, and xylene-induced ear edema methods. Antioxidant potential was implemented by the DPPH scavenging method. The SIF-ME consistently displayed significant anti-nociceptive activity in a dose-dependent pattern (p < 0.05). The maximum analgesic activity was found in the highest dose (200 mg/kg; p < 0.001) in a hot plate, acetic acid-induced writhing test 43.47%, and in formalin-induced licking test in both early phase (43.3%; p < 0.01) and late phase (61.84%; p < 0.001%). The extract provided optimal protection against hemolysis (83.41% decrease) at 1000 µg/mL and significantly inhibited protein denaturation (67.34-26.05%) at doses of 1000-62.5 µg/mL. At 200 mg/kg, the extract showed dose-dependent and substantial inhibition (54.07%; p < 0.01) of xylene-induced ear edema. The in-vitro DPPH (IC50 = 469.5 µg/mL) results showed remarkable scavenging activity and concentration-dependent reducing power. The extract demonstrates no acute oral toxicity, as indicated by an LD50 value exceeding 1000 mg/kg body weight. Gas Chromatography-Mass Spectrometry/Mass Spectrometry (GC-MS/MS) analysis was performed which yielded sixty bioactive compounds. In-silico and molecular docking studies revealed favorable pharmacological properties, including good binding affinities and ADME/T profiles. These results support the medicinal use of the plant, which makes it a potential source of analgesic, anti-inflammatory, and antioxidant candidates.

Characterization of the bioactive components of Chloris virgata Sw. and assessment of its antimicrobial and anti-arthritic potential

The goals of this study are to characterize the bioactive ingredients present in the formulation of three parts of Chloris virgata Sw. and to assess their antimicrobial and anti-arthritic properties. C. virgata Sw. extracts (methanol, hydro-ethanol, and aqueous) were characterized using ultraviolet-visible (UV-Vis) spectroscopy and Fourier transform infrared spectroscopy (FTIR) techniques. Antimicrobial activity was measured with the disc diffusion method, and anti-arthritic activity was determined by an in vitro protein denaturation assay. The in-vivo study was performed on methanol extract, and the biochemical and hematological parameters were investigated by Freund’s complete adjuvant model. UV-visible spectroscopy revealed the presence of flavonoids, terpenoids, phenols, alkaloids, and saponins, whereas FTIR results show functional groups like carboxylic acids, alkenes, amides, carbohydrates and proteins, ether, and esters. Methanol and hydroethanol extracts showed higher antimicrobial activity at 400 μg/mL against Escherichia coli, Bacillus subtilis, and Aspergillus niger in comparison to the standards streptomycin and fluconazole at 10 μg/ mL. In-vitro results of the methanol extract clearly show 72.58% inhibition as compared to other extracts. In-vivo anti-arthritic evaluation was performed with methanol extract, and it showed remarkable paw reduction in the treated group on 21st day at a high dose (2.2 ± 0.1) as compared to indomethacin (1.74 ± 0.114). Hematological parameters such as hemoglobin, red blood cells (RBC), white blood cells (WBC), neutrophils, lymphocytes, and platelets, and biochemical parameters like alkaline phosphatase (ALP) (51.36 ± 9.113 IU/L), aspartate aminotransferase (AST) (64.65 ± 10.011 IU/L), and alanine aminotransferase (ALT) (129.35 ± 13.53 IU/L) were restored to normal in the treated. The findings indicated that C. virgata extracts under UV spectroscopy and FTIR revealed significant bioactive components, which are determined to be significant contributors to their antioxidant and antibacterial properties.

Anti-inflammatory and antioxidant effects of hydro-alcoholic extract of Dicliptera cuneata Nees aerial parts.

Dicliptera cuneata Nees is a traditional medicinal plant but its extract or phytochemicals are less known. Therefore, it is of interest to investigate the anti-inflammatory and antioxidant effects of aerial part hydroalcoholic extract of Dicliptera cuneata Nees. Hence, we used protein denaturation assay, FRAP assay, Nitric oxide and peroxide scavenging assays methods following standard developed techniques. The hydro-alcoholic extract exhibited dose-dependent effectiveness in all the assays and showed maximum efficacy in the assays at higher doses selected. Data shows that hydroalcholic extract of Dicliptera cuneata Nees showed anti-inflammatory and antioxidant properties in in vitro settings. It should be noted that more data is needed to further develop the extract into suitable formulations.

An in vitro study for the evaluation of morphological and biochemical characteristics of absorbable sutures coated with genistein and nicotine

Introduction and aim. Inflammation, cell proliferation, matrix deposition, and tissue remodeling are all elements of the well-structured and well-coordinated process of wound repair. The aim of this study was to analyze the effect of genistein and nicotine on polyglycolic acid (PGA) and vicryl sutures. Material and methods. Genistein and nicotine were isolated and solution was prepared and the suture material PGA and vicryl were immersed in the solution and dried. They were tested for their tensile strength and degradation values after immersion in artificial saliva (on the first day and on the 14th day). The sutures were also seen under a scanning electron microscope (SEM) for its uniform coating and the mixture formulation of genistein and nicotine were tested for their anti-inflammatory and antioxidant activity using protein denaturation assay and 2,2-diphenyl-1-picrylhydrazyl assay respectively. Results. Nicotine has a high anti-inflammatory activity on the suture material, whereas Genistein has an insignificant anti-inflammatory effect. The mixture formulation has a relatively similar anti-inflammatory effect when compared to the control. The SEM analysis shows a uniform coating of the formulation on the PGA and vicryl sutures. In comparison, PGA has shown lesser tensile strength and hence higher degradation ability. Conclusion. Nicotine and Genistein affect the tensile strength and degradation properties of the sutures.

Molecular structure, characterization, in vitro and in-silico studies of N,N-dimethyl aminophenyl schiff's base-chalcone hybrid

This study reports the synthesis and characterization of a novel compound, a chalcone-based Schiff base (SBC), with potential applications in anti-inflammatory and anti-diabetic treatments. Chalcones, known for their diverse biological activities, are combined with Schiff bases, versatile compounds, to create the SBC. The synthesis process involves aldol condensation and Schiff base formation. The compound's structure is confirmed through NMR, FT-IR, and mass spectroscopic analyses. The UV–visible spectroscopy reveals the compound's electronic properties, and molecular docking experiments indicate its potential to interact with enzymes related to inflammation and diabetes. Anti-inflammatory activity is evaluated through a protein denaturation assay using bovine serum albumin. SBC demonstrates significant anti-inflammatory activity, outperforming standard drug molecules, and exhibits concentration-dependent effects. In an α-amylase inhibitory assay, a vital enzyme in the digestive process related to diabetes, SBC shows remarkable inhibition, surpassing standard drug performance, especially at higher concentrations. Molecular docking studies suggest the compound's potential interactions with enzymes responsible for inflammation and diabetes. SBC displays strong binding energies and inhibition constants with α-amylase, indicating its suitability for diabetes-related studies. The synthesis and characterization of SBC, combined with its excellent anti-inflammatory and anti-diabetic activities, underscore its potential as a promising candidate for further biomedical research and therapeutic development.

Synthesis of Silver Nanoparticles From Cymodocea rotundata Leaf Extract and Their Biological Activities.

Silver nanoparticles (AgNPs) are considered to be a very significant and intriguing type within the category of metallic nanoparticles, particularly in the context of their involvement in biological applications. The objective of this research is to use the green synthesis method in order to synthesize AgNPs by using the leaf extract of C. rotundata. Furthermore, the study aims to evaluate the antioxidant and anti-inflammatory properties of these nanoparticles. Fresh and healthy specimens of C. rotundata were gathered from Palk Bay, Tamil Nadu, India, and afterward subjected to a thorough washing process using tap water. The cleaned materials were air-dried and then fragmented into small bits and finely ground. The ethanolic extract of seagrass was then combined with a solution containing 1 millimolar (mM) silver nitrate (AgNo3). The decrease of silver ions in the solution was frequently measured using a UV-visible spectrophotometer.Synthesized AgNPs were investigated for antioxidants by DPPH (2,2-diphenyl-1-picrylhydrazyl) assay and anti-inflammatory activity was measured by protein-denaturation assay. The use of C. rotundata leaf extract in the green synthesis of AgNPs, in the presence of 1 mM AgNO3, led to a noticeable alteration in the colour of the mixture, transitioning from a pale hue to a brown shade. This change in colour serves as evidence of the reduction of AgNo3 ions to silver ions, thereby facilitating the creation of AgNPs. The duration of the bio-reduction process of silver ions in the reaction mixture was observed to be two hours. The antioxidant and anti-inflammatory activity showed promising activity for AgNPs. This study concluded that C. rotundata had antioxidant capabilities, and AgNPs derived from C. rotundata have potential use in pharmaceuticals and medication administration.

Anti-Inflammatory Property of Costus afer Ker Gawl Ethanol Leaf Extract in STZ-Induced Diabetic Rats

Costus afer Ker Gawl is an indigenous plant, commonly called a ginger lily, spiral ginger, or bush cane, it has been reported to possess anti-inflammatory properties. The anti-inflammatory activity of C.afer leaf extract in streptozotocin-induced diabetic rats was investigated. Protein denaturation and erythrocyte stabilization assays were used to evaluate in vitro anti-inflammatory activity, and alpha-amylase and alpha-glucosidase enzyme inhibition was used to evaluate in vitro anti-diabetic activity. 60 male Wistar rats were used for the two inflammatory models: the 30 rats were randomly assigned into six groups (n=5) for carrageenan-induced paw edema and cotton pellet granuloma models respectively: Group I: normal, group II: control(untreated), group III: 10mg/kg b.w diclofenac sodium (standard), group IV, V and VI were given 50, 100, 250 mg/kg b.w Costus afer ethanol leaf extract (CAELE) in each of the two models. The study showed that in protein denaturation assay, CAELE and Diclofenac had 56.69% and 80.82% respectively at the highest concentration, erythrocyte stabilization had 80.40% CAELE and 94.88% Diclofenac sodium at the highest dose in a dose-dependent manner. Alpha amylase and alpha-glucosidase showed an increase in percentage inhibition activity at 65.44% and 43.72% respectively against acarbose (standard) at 56.01%. However, in the cotton pellet-induced granuloma model, the concentration exhibited high percentage inhibition (77.82%) comparable to the standard drug at 91.28%, and reduction in paw thickness was also observed in the carrageenan model in a dose-dependent manner respectively. This study showed that CAELE at different concentrations showed anti-inflammatory activity in diabetic conditions.

In-vitro Pharmacological Investigation of Ludwigia adscendens

Aims: This research objective was to investigate some in-vitro properties of methanol-extracted plant extracts of Ludwigia adscendens (the water primrose). The primary method of assessment for the methanolic extract of Ludwigia adscendens (MELA) was investigated for phytochemical screening. It was determined to look into the plant's potential for in vitro activities, such as antioxidant, antimicrobial, anti-inflammatory anti-arthritis, cytotoxic, thrombolytic and membrane stabilizing activity due to the pharmaceutical interest in its component parts.&#x0D; Methodology: Phytochemical screening was performed utilizing a battery of test reagents. Antioxidant, antimicrobial, anti-inflammatory, anti-arthritic, cytotoxic, thrombolytic and membrane stabilizing activities were measured at various doses using the DPPH Free Radical Scavenging Assay, the Disk Diffusion Method, Egg Albumin Protein Denaturation Assay, the Inhibition of Protein Denaturation Assay, the Brine Shrimp Lethality Bioassay, Clot lysis assay and heat induced and hypotonic induced haemolysis method.&#x0D; Results: The findings of research on MELA's phytochemical screening showed the presence of a wide range of various phytochemical components. The higher antioxidant activity of MELA is shown by its IC50 value of 85.76 µg/mL. Significant antibacterial action was shown by MELA. As compared to the standard ascorbic acid, this extract showed significantly greater anti-arthritic and anti-inflammatory effects. MELA exhibited much higher LC50 value which is 1.147 µg/mL than the typical vincristine-sulphate in a lethality test with brine shrimp. Comparatively speaking, this herb has quite powerful thrombolytic action. Significant membrane stabilizing action has also been shown by MELA.&#x0D; Conclusion: To wrap it up, it is clear that this plants phytochemical can be used for wide range of drug discovery field due to its potent pharmacological actions.

Comparative study of phytochemicals, antioxidant and anti-inflammatory activity of infusion and decoction extracts from Pycnoporus sanguineus (L.) Murril collected in West Bengal

Mushrooms are well-known pharmaceutical agents as they possess bioactive secondary metabolites along with a large amount of proteins, carbohydrates, crude fiber, fat and other beneficial compounds. Due to their remarkable pharmacological property, they are used in traditional medicine in different countries of Asia. Pycnoporus sanguineus is a polypore fungus having a wide variety of bioactive substances and therapeutic activities that are reported previously in different countries of the world. The present study is focused to estimate different bioactive compounds and evaluate antioxidant and anti-inflammatory activities of P. sanguineus using two different extraction procedures (infusion and decoction). In this study, total phenol, total flavonoid, β-carotene, lycopene and ascorbic acid contents are estimated. Antioxidant and inti-inflammatory activities are investigated by total antioxidant capacity, DPPH radical scavenging, ABTS radical scavenging, chelating of ferrous ion (Fe2+), reducing power assays and protein denaturation assay respectively. The aim of the study is to establish P. sanguineus as a natural drug that could be used against oxidative stress and inflammation-related diseases substituting commercial drugs.

A plant protein farnesylation system in prokaryotic cells reveals Arabidopsis AtJ3 produced and farnesylated in E. coli maintains its function of protecting proteins from heat inactivation

BackgroundProtein farnesylation involves the addition of a 15-carbon polyunsaturated farnesyl group to proteins whose C-terminus ends with a CaaX motif. This post-translational protein modification is catalyzed by a heterodimeric protein, i.e., farnesyltransferase (PFT), which is composed of an α and a β subunit. Protein farnesylation in plants is of great interest because of its important roles in the regulation of plant development, responses to environmental stresses, and defense against pathogens. The methods traditionally used to verify whether a protein is farnesylated often require a specific antibody and involve isotope labeling, a tedious and time-consuming process that poses hazardous risks.ResultsSince protein farnesylation does not occur in prokaryotic cells, we co-expressed a known PFT substrate (i.e., AtJ3) and both the α and β subunits of Arabidopsis PFT in E. coli in this study. Farnesylation of AtJ3 was detected using electrophoretic mobility using SDS-PAGE and confirmed using mass spectrometry. AtJ3 is a member of the heat shock protein 40 family and interacts with Arabidopsis HSP70 to protect plant proteins from heat-stress-induced denaturation. A luciferase-based protein denaturation assay demonstrated that farnesylated AtJ3 isolated from E. coli maintained this ability. Interestingly, farnesylated AtJ3 interacted with E. coli HSP70 as well and enhanced the thermotolerance of E. coli. Meanwhile, AtFP3, another known PFT substrate, was farnesylated when co-expressed with AtPFTα and AtPFTβ in E. coli. Moreover, using the same strategy to co-express rice PFT α and β subunit and a potential PFT target, it was confirmed that OsDjA4, a homolog of AtJ3, was farnesylated.ConclusionWe developed a protein farnesylation system for E. coli and demonstrated its applicability and practicality in producing functional farnesylated proteins from both mono- and dicotyledonous plants.

Evaluation of In-vitro Antimicrobial, Anti-arthritis and In-vivo Analgesic and Neuropharmacological Investigation of the Bark Extract of Solanum americanum Milli

Purpose: The objective of this study was to investigate the impact of methanol-prepared bark extract of Solanum americanum Milli. Bark on several in vitro activities, including antimicrobial and anti-arthritis effects. Additionally, the study aimed to assess the analgesic and neuropharmacological properties of the extracts in an animal model.&#x0D; Place and Duration of Study: The research study was carried out from March 2023 to July 2023 at the Bangladesh Council of Scientific and Industrial Research (BCSIR) and the Laboratory of Phytochemistry and Pharmacology, Department of Pharmacy, as well as the Laboratory of Microbiology at Stamford University Bangladesh, Dhaka.&#x0D; Methodology: The methodology involved various doses of methanolic bark extract of Solanum americanum Milli (MESAB), and employed techniques such as the Disk Diffusion Method for antimicrobial test and the Inhibition of Protein Denaturation Assay for anti-arthritic test. The analgesic efficacy was assessed by the hot plate test and administration of acetic acid induced writhing responses in Swiss albino mice. The mechanism test is conducted through the cyclic guanosine monophosphate (cGMP) test. The neuropharmacological inquiry included the measurement of locomotor activity through the utilization of open field and hole cross tests.&#x0D; Results: The Disk Diffusion Test of MESAB yielded findings indicating its antimicrobial properties against all strains used in this research. The findings from the study examining the anti-arthritis properties indicate that MESAB exhibits a significant efficacy of 94.59% in inhibiting arthritis at a concentration of 1000 µg/mL. This level of efficacy is comparable to that of diclofenac sodium, which also has a similar efficacy of 98.19% at the same concentration. The findings of the present study demonstrated that MESAB displayed analgesic properties in both analgesic models, suggesting the involvement of both central and peripheral pathways. Based on the results of the study, it was observed that the administration of MESAB at a dosage of 400 mg/kg had a significant analgesic effect. The application of MB before to treatment resulted in a further enhancement of this effect, leading to an increase in the percentage of inhibition from 66.75% to 79.70%, respectively. The consideration of the involvement of the GABA-benzodiazepine receptor in neuropharmacological activity holds significant importance. In the Open Field and Hole Cross Tests, it was shown that the extract exhibited significant result compared to Diazepam in terms of increasing motor coordination.&#x0D; Conclusion: In the future, plant-derived pharmacological compounds may offer potential benefits for various clinical disorders, such as neurodegenerative illnesses. These substances possess neuropharmacological properties, antibacterial effects, and anti-arthritic activity, which could potentially serve as alternatives to non-steroidal anti-inflammatory drugs (NSAIDs).

Fagopyrum tataricum (L.) Gaertn interacts with Gsk-3β/Nrf-2 signalling to protect neurotoxicity in a zebrafish model

Ethnopharmacological relevanceFagopyrum tataricum (L.) Gaertn is used as a folk medicine in many Asian countries due to its anti-inflammatory, antioxidant, and several other health-promoting properties. It is also prescribed to improve neurocognitive functions and alleviate inflammatory conditions. Aim of the studyOxidative stress and neuroinflammation plays a crucial role in neurodegenerative conditions. Hence, based on the ethnomedical claims and available literature, the present study investigated neuroprotective efficacy of a seed extract (ft-ext) of Fagopyrum tataricum against acrylamide (ACR)-induced neurotoxicity. Materials and methodsThe phytochemical characterization of ft-ext was performed by a high-performance liquid chromatography method. Molecular interactions of the identified compounds of ft-ext were studied using an in-silico docking tool. An in-vitro protein denaturation assay was done to check anti-inflammatory activity. The 5 days’ post-fertilized zebrafish larvae were exposed to 1 mM and 2.5 mM ACR with or without ft-ext for 72 h to study its neuroprotective efficacy. Real-time polymerase chain reaction and western blotting studies were performed to analyse the oxidative stress-related gene and protein expressions respectively. ResultsThe extract showed the presence of chlorogenic acid, rutin, caffeic acid, vitexin, syringic acid, quercetin, p-coumaric acid, kaempferol, and ferulic acid. In-vitro protein denaturation assay of ft-ext showed a potent anti-inflammatory effect. The ft-ext improved ACR-mediated locomotor deficit and reduced overall mortality in the larvae. The brain lipid peroxidation and protein carbonylation results revealed an elevated level of oxidative stress in the ACR-treated group, which was reduced in ft-ext-treated larvae. The extract treatment increased the expression of nrf2, gpx, and hmox1a, while simultaneously downregulated trxr2 levels in the brain of larvae exposed to ACR. The treatment also showed inactivation of Gsk-3β, thus maintaining a normal pool of Nrf2 and β-catenin. Molecular docking of identified compounds of ft-ext showed possible hydrogen and hydrophobic interactions with Gsk-3β. ConclusionThe ft-ext prevents ACR-mediated neurotoxicity by suppressing Gsk-3β mediated oxidative stress.

Design, Synthesis, Characterization and Non-steroidal Anti-Inflammatory Activity of Novel Oxazolone Derivatives

An innovative sequence of Oxazolone derivatives were synthesized via chemical reaction including Preparation of benzoyl glycine crystals and Synthesis of oxazolone derivatives. The synthesized compounds such as P- Chlorobenaldehyde, 4-Fluro benaldehyde, Cinnamaldehyde, 2-Chloro benaldehyde, and O- Anisaldehyde were elucidated by Fourier Transform-Infra Red (FT-IR) spectrometer and showed its corresponding peaks. The in- vitro anti-inflammatory activity of synthesized compounds by protein denaturation assay exhibited very good significant anti-inflammatory activity. Additionally, the test compounds 2-Chloro benzaldehyde had nearest IC50 value to the standard diclofenac sodium when compared to all other test compounds, and demonstrated highest zone of inhibition at the low concentration used to exhibit the high potency.

In vitro anti-inflammatory and anti-ulcer activities of aqueous extract of Emilia sonchifolia

Emilia sonchifolia, an annual herb belonging to the Asteraceae family, has long been recognized for its potential in treating cutaneous infections, inflammation, and wounds. This study aimed to assess the anti-inflammatory and anti-ulcer effects of Emilia sonchifolia using protein denaturation assay and Acid Neutralizing Capacity (ANC) methods. The acid neutralizing capacity method revealed that the extract significantly reduced acidity to 8.35 at a concentration of 1000 mg/ml, outperforming the standard Aluminum hydroxide + Magnesium hydroxide combination which achieved a value of 13.2 at 500 mg/ml. Furthermore, the results from the protein denaturation assay demonstrated a positive correlation between increased concentration and percentage inhibition of inflammation. The extract exhibited the highest inhibition percentage of 44.99 at a concentration of 200 µg/ml, and the standard diclofenac sodium achieved 84.120 at 100 µg/ml. These findings underscore the remarkable medicinal potential of aqueous extract of Emilia sonchifolia, particularly in its applications as an anti-inflammatory and anti-ulcer agent. In summary, the aqueous extract of Emilia sonchifolia demonstrated significant anti-inflammatory activity across various concentrations, when compared to the effectiveness of the standard drug diclofenac sodium. Additionally, the extract displayed anti-ulcer activity when compared to the standard Aluminum hydroxide + Magnesium hydroxide combination. Phytochemical analysis unveiled the presence of alkaloids, glycosides, saponins, flavonoids, terpenoids, phenolic compounds, steroids, and carbohydrates in Emilia sonchifolia. These phytochemical constituents likely contribute to the extract's protective properties against protein denaturation and its anti-ulcer effects.

In vitro Antioxidant and Anti-Inflammatory Effects of the Hydro-Methanolic Extract of Phlomis crinita from North Algeria

Abstract: Phlomis crinita is a medicinal plant commonly used in Algerian folk medicine as a remedy for ulcers, haemorrhoids and healing of wounds. The current study aimed to evaluate the anti-inflammatory and antioxidant effects of this plant and to provide a scientific basis for its use. Protein denaturation and membrane stabilization assays were used to assess the anti-inflammatory action in vitro. DPPH free radical-scavenging and reducing power tests were used to determine the antioxidant activity. The plant methanol extract exhibited potential anti-inflammatory activity, demonstrating its ability to prevent protein denaturation and stabilize the membrane of human red blood cells. It had IC50 values of 143.64 ± 2.94 and 119.63 ± 0.80 µg/mL, respectively, compared to diclofenac sodium (IC50 value of 60.60 ± 2.55 and 53.47 ± 0.93 µg/mL, respectively). P. crinita showed strong antioxidant DPPH scavenging activity (IC50 = 82.07±0.37 μg/mL) compared to ascorbic acid (62.48±0.56) and exhibited a strong reducing potential. The results suggested that the plant might be a potential candidate for oxidative stress prevention and inflammatory disease treatment.

Pharmacological Investigation of In-vitro Anti-inflammatory, Antimicrobial, Thrombolytic, Cytotoxic and In vivo Analgesic Activities of Ethanolic Leaf Extract of Diospyros malabarica

Aims: This study was designed to investigate the effects of leaf extracts of Diospyros malabarica plant (EEDML) on a number of in vitro activities, including antibacterial, anti-inflammatory, cytotoxic, and thrombolytic activities, as well as in vivo analgesic activities.&#x0D; Place and Duration of Study: The Laboratory of Phytochemistry and Pharmacology at the Department of Pharmacy and the Laboratory of Microbiology at Stamford University Bangladesh and the Bangladesh Council of Scientific and Industrial Research, Dhaka, conducted this investigation from January 2022 to June 2023.&#x0D; Methodology: The study employed the Egg Albumin Protein Denaturation Assay to evaluate the anti-inflammatory effects of various doses. The Disk Diffusion Method was employed for conducting an antimicrobial assay. Different tests were conducted to evaluate the effects of thrombolytic and cytotoxic substances. These tests included the Clot Lysis assay and the Brine Shrimp Lethality Assay. In-vivo analgesic testing was conducted using the hot plate method and the acetic acid-induced writhing test.&#x0D; Results: The experimental tests showed significant pain-relieving effects, with inhibition percentages of 46.2% and 66.61% observed at doses of 200 and 400 mg/kg, respectively. The anti-inflammatory test showed a significant inhibition of EEDML at a concentration of 1000μg/mL, with an inhibition rate of 85.45%. The antimicrobial test showed a moderate antimicrobial effect, with a zone of inhibition ranging from 07-15, depending on the microorganism and dosage. The thrombolytic test showed a clot lysis rate of 85.37%, while the cytotoxic test indicated an LC50 value of 1.47.&#x0D; Conclusion: EEDML's phytochemical displays strong pharmacological properties, suggesting its potential for drug discovery in diverse areas.

Phytochemical profiling of Salsola tetragona Delile by LC-HR/MS and investigation of the antioxidant, anti-inflammatory, cytotoxic, antibacterial and anti-SARS-CoV-2 activities

This study aimed to investigate the phytochemical composition and biological activity of Salsola tetragona Delile. (Amaranthaceae), a medicinal plant. The study evaluated the antioxidant potential of the crude extract and five fractions of S. tetragona using DPPH•, ABTS•+, CUPRAC, and metal chelating assays. The anti-inflammatory activity was determined using a protein denaturation assay, and the antibacterial activity was determined by the Minimum inhibitory concentrations (MICs) for the growth of Escherichia coli and Staphylococcus aureus strains. The MTT test and an in vitro scratch assay evaluated the effects on cell viability and cell migration. The potential anti-SARS-CoV-2 activity was assessed by analyzing the effects on the interaction between ACE2 and Spike protein. The bioactive compounds present in the plant were identified using LC-HR/MS analysis. The crude hydromethanolic extract (STM) and five fractions of S. tetragona, n-hexane (STH), dichloromethane (STD), ethyl acetate (STE), n-butanol (STB), and aqueous (STW) showed significant antioxidant activity in four different tests. In the anti-inflammatory assay, the ethyl acetate fraction exhibited significantly higher activity than Aspirin® (IC50 = 13 ± 5 µg/mL). The crude extract and its fractions showed positive antibacterial activity with similar MICs. In the cytotoxicity assay against the breast cancer cell line MCF7, the dichloromethane fractions (STD) were very effective and demonstrated superiority over the other fractions (IC50 = 98 µg/mL). Moreover, the potential of the extract and fractions as anti-SARS-CoV-2, the ethyl acetate, and dichloromethane fractions demonstrated important activity in this test. LC-HR/MS analysis identified 16 different phenolic compounds, Eleven of which had not been previously reported in the genus Salsola. The results suggest that the extracts of S. tetragona have the potential to become new sources for developing plant-based therapies for managing a range of diseases.