Free Radical Scavenging, Anti-inflammatory and Antibacterial Activity of Acorus calamus Leaves Extract Against Pseudomonas aeruginosa and Staphylococcus aureus.

Background Herbal medicine, or phytotherapy, has been used for centuries in traditional healing practices to harness the therapeutic properties of different plant-derived elements. Acorus calamus, a perennial herbaceous plant, has significant historical importance in traditional medicine, specifically in Ayurveda, where it is referred to as "Vacha." This study investigates the antioxidant, anti-inflammatory, and antimicrobial characteristics of the A. calamus dimethyl sulfoxide (DMSO) extract. The objectives of the research are to provide valuable knowledge about the preparation of A. calamus DMSO extract and to explore its potential anti-inflammatory, antioxidant, and antimicrobial effects. Materials and methods The A. calamus DMSO extract was derived from leaves, and its antioxidant activity was evaluated through the use of the 2,2-diphenyl-1-picryl hydrazyl (DPPH) assay, hydroxyl radical scavenging assay (H2O2 assay), and ferric reducing antioxidant power (FRAP) assay. The anti-inflammatory activity was assessed using the Bovine serum albumin (BSA) denaturation assay, egg albumin (EA) denaturation assay, and membrane stabilization assays. The antimicrobial activity was analyzed using the agar well diffusion technique and the time-kill curve assay. Results In DPPH and H2O2 tests, the DMSO extract of A. calamus showed significant antioxidant activity, near that of standard ascorbic acid. The FRAP assay demonstrated a correlation between the dose and the activity of reducing ferric ions. The A. calamus DMSO extract exhibited significant anti-inflammatory properties in BSA and EA denaturation assays, similar to the standard diclofenac sodium. The anti-inflammatory potential of the A. calamus DMSO extract was further confirmed through the membrane stabilization assay. The DMSO extract of A. calamus exhibited a significant inhibition zone against the pathogens Streptococcus mutans and Pseudomonas aeruginosa during the antimicrobial evaluation, surpassing the efficacy of the standard antibiotic. The time-kill curve assay validated the antibacterial efficacy, which was dependent on the concentration. Conclusion The A. calamus DMSO extract exhibited promising antioxidant, anti-inflammatory, and antimicrobial properties, supporting its traditional use in alternative medicine. The findings suggest its potential as a natural resource of compounds with bioactive properties for use in pharmaceutical and nutraceutical applications.

Phytochemical analysis and biological investigation of Cheilanthes tenuifolia (Burm.f.) Swartz.

Introduction: Cheilanthes tenuifolia is an evergreen ornamental small fern, belonging to the family Pteridaceae, that grows in warm and rocky regions worldwide. Many species of Cheilanthes genus are evidently endowed with important phytochemicals and bioactivities. This study aimed to perform a preliminary phytochemical analysis of Cheilanthes tenuifolia leaves alongside an evaluation of free radical scavenging, anti-inflammatory, antimicrobial, and clot lysis activities of extract fractions. Materials and methods: A preliminary phytochemical analysis was done after fractionation of ethanolic extract (ECT) with n-hexane (HCT) and chloroform (CCT). Then, 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging, egg albumin and RBC membrane stabilization tests, disc diffusion, and human blood clot lysis assays were performed. Results: Phytochemical investigations suggested that the plant is rich in alkaloids, glycosides, tannins, and flavonoids. All obtained fractions exhibited concentration-dependent radical scavenging, inhibition of egg protein denaturation and RBC membrane lysis capacities. Except for antifungal tests, ECT exhibited better DPPH radical scavenging, anti-inflammatory, antibacterial, and clot lysis capacities than HCT and CCT fractions. However, all fractions exhibited a mild anti-inflammatory activity. Conclusion: C. tenuifolia might be a good source of antioxidant, anti-microbial, and anti-atherothrombotic agents. Further studies are required to isolate and characterize the active principles liable for each bioactivity, along with possible molecular interactions.

In-vitro and In-silico Assessment of Anti-inflammation Properties of Saponarin Extracted from Hordeum Vulgare.

BACKGROUND: Hordeum vulgare, commonly known as Barley grass, is a historically significant cultivated plant with profound implications for societies, agricultural sciences, and human nutrition. It has been valued for both sustenance and its potential medicinal properties. OBJECTIVE: This study aims to comprehensively investigate the medicinal properties of Hordeum vulgare, focusing on its potential therapeutic benefits and anti-inflammatory properties. Additionally, we seek to quantify and compare the phytochemical content of two distinct extracts: Barley Grass Hexane Extract (BGHE) and Barley grass aqueous extract (BGAQ). METHODS: We quantified the phytochemical contents of BGHE and BGAQ and evaluated their antiinflammatory effects using UV spectroscopy at 560 nm, coupled with the RBC membrane stabilization technique. Subsequently, we conducted in-silico studies to assess the in vitro anti-inflammatory potential of Barley grass leaf extracts. RESULTS: Both BGHE and BGAQ demonstrated significant inhibitory effects on inflammation compared to the control group. However, BGHE exhibited superior anti-inflammatory efficacy when compared to BGAQ, suggesting its role as a potential anti-inflammatory agent. In-silico studies further supported the anti-inflammatory potential of Barley grass leaf extracts. CONCLUSION: Hordeum vulgare, or Barley grass, offers a wealth of health benefits, including antiinflammatory, anti-diabetic, anti-cancer, antioxidant, anti-acne, and anti-depressant properties. These properties contribute to improved immunity, reduced cardiovascular disorders, and alleviation of fatigue. The distinct extracts, BGHE and BGAQ, both exhibit promising anti-inflammatory capabilities, but BGHE shows better anti-inflammatory activity. This research sheds light on the therapeutic potential of Barley grass, making it a valuable candidate for further exploration in the field of natural medicine.

Simultaneous Study of Analysis of Anti-inflammatory Potential of Dryopteris ramosa (C. Hope) C. Chr. using GC-Mass and Computational Modeling on the Xylene-induced Ear Oedema in Mouse Model.

INTRODUCTION: In the present study, we aimed to investigate the extraction and identification of the potential phytochemicals from the Methanolic Extract of Dryopteris ramosa (MEDR) using GC-MS profiling for validating the traditional uses of MEDR its efficacy in inflammations by using in-vitro, in-vivo and in silico approaches in anti-inflammatory models. METHODS: GC-MS analysis confirmed the presence of a total of 59 phytochemical compounds. The human red blood cells (HRBC) membrane stabilization assay and heat-induced hemolysis method were used as in-vitro anti-inflammatory activity of the extract. The in-vivo analysis was carried out through the Xylene-induced mice ear oedema method. It was found that MEDR at a concentration of 20 µg, 30 µg, and 40 µg showed 35.45%, 36.01%, and 36.33% protection to HRBC in a hypotonic solution, respectively. At the same time, standard Diclofenac at 30 µg showed 45.31% protection of HRBC in a hypotonic solution. RESULTS: The extract showed inhibition of 25.32%, 26.53%, and 33.31% cell membrane lysis at heating at 20 µg, 30 µg, and 40 µg, respectively. In comparison, standard Diclofenac at 30 µg showed 50.49% inhibition of denaturation to heat. Methanolic extract of the plant exhibited momentous inhibition in xylene-induced ear oedema in mice treated with 30 µg extract were 47.2%, 63.4%, and 78.8%, while inhibition in mice ear oedema treated with 60 µg extract was 34.7%, 43.05%, 63.21% and reduction in ear thickness of standard drug were 57.3%, 59.54%, 60.42% recorded at the duration of 1, 4 and 24 hours of inflammation. Molecular docking and simulations were performed to validate the anti-inflammatory role of the phytochemicals that revealed five potential phytochemicals i.e. Stigmasterol,22,23dihydro, Heptadecane,8methyl, Pimaricacid, Germacrene and 1,3Cyclohexadiene,_5(1,5dimethyl4hexenyl)-2methyl which revealed potential or significant inhibitory effects on cyclooxygenase-2 (COX-2), tumour necrosis factor (TNF-α), and interleukin (IL-6) in the docking analysis. CONCLUSION: The outcome of the study signifies that MEDR can offer a new prospect in the discovery of a harmonizing and alternative therapy for inflammatory disease conditions.

Exploring therapeutic potential of Woodfordia fruticosa (L.) Kurz leaf and bark focusing on antioxidant, antithrombotic, antimicrobial, anti-inflammatory, analgesic, and antidiarrheal properties.

Background and Aims: The study aimed to evaluate the pharmacological properties of methanolic extracts of leaves and barks of Woodfordia fruticosa (L.) Kurz (family: Lythraceae) focusing on antioxidant, thrombolytic, anti-inflammatory, antibacterial, analgesic, and antidiarrheal effects. Methods: 1,1-Diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging assay, clot lysis, disc diffusion, and membrane stabilizing methods were employed to assess in vitro antioxidant, thrombolytic, antibacterial, and anti-inflammatory properties of the leaf and bark methanolic extracts (ME) of W. fruticosa and different organic solvents, that is, petroleum ether (PE), dichloromethane (DCM), chloroform (CL), and aqueous (AQ) fractions. In addition, in vivo central and peripheral analgesic and antidiarrheal activities of both crude extracts were evaluated at two doses (200 and 400 mg/kg of body weight [bw]). Results: All the extracts and fractions showed promising antioxidant properties by scavenging DDPH free radicals with IC50 of 6.11-20.79 µg/mL. AQ fraction (41.24%) of leaves and ME (44.90%) of bark exerted notable in vitro thrombolytic activity. The CL fraction of leaves and AQ fraction of the bark showed 43.16% and 45.37% inhibition of RBC hemolysis, respectively, compared to the inhibition of RBC hemolysis by aspirin in a hypotonic-induced membrane stabilizing assay. Besides, both extracts were observed to provide significant (p < 0.001) central and peripheral analgesic responses at both doses of 200 and 400 mg/kg bw. Furthermore, both doses of bark extract (p < 0.001) and the 400 mg/kg bw of leaf extract (p < 0.05) were observed to possess statistically significant antidiarrheal activity. Additionally, in an in vivo acute toxicity investigation, both extracts had a median lethal dose (LD50) greater than 5000 mg/kg bw, indicating their safety level. Conclusion: The current study proves the ethnomedicinal uses of W. fruticosa; however, further studies are required for phytochemical screening to isolate the responsible bioactive compounds and discover the lead molecules from the plant species.

Pentaclethra eetveldeana Leaves from Four Congo-Brazzaville Regions: Antioxidant Capacity, Anti-Inflammatory Activity and Proportional Accumulation of Phytochemicals.

Oxidative stress and inflammation play a key role in the occurrence of neurodegenerative diseases. Traditionally, Pentaclethra eetveldeana leaves are used in dementia treatment. Therefore, this study aimed to evaluate the antioxidant and anti-inflammatory activities as well as the phytochemical composition of Pentaclethra eetveldeana leaves from four Congo-Brazzaville regions. The 1.2-diphenyl-1-picrylhydrazyl radical-scavenging, ß-carotene bleaching and molybdenum reduction assays were used to assess the antioxidant activity. The protein denaturation and erythrocyte membrane stabilization tests were used to analyze the anti-inflammatory activity. Phytochemical screening, the quantification of polyphenols by spectrophotometry, as well as the determination of extraction yields were carried out. It was found that the extracts reduced molybdenum; furthermore, compared to ascorbic acid, they showed better antiradical activity and inhibited lipid peroxidation. Moreover, globally, the membrane-stabilizing power of the aqueous extracts was superior or comparable to diclofenac, while the same extracts were less effective for the inhibition of denaturation. All of the aqueous extracts contained polyphenols, saponins, alkaloids, anthraquinones, reducing sugar and cardiotonic glycosides. The total polyphenols, tannins and proanthocyanidins are produced proportionally from one region to another. Finally, the leaves from Brazzaville and Boundji contain flavonols, while those from Makoua and Owando contain flavones. Thus, Pentaclethra eetveldeana leaves contribute to traditional dementia treatment through their antioxidant and anti-inflammatory properties.

Insight into the biochemical and cell biological function of an intrinsically unstructured heat shock protein, Hsp12 of Ustilago maydis.

Small heat shock proteins (sHsps) play diverse roles in the stress response and maintenance of cellular functions. The Ustilago maydis genome codes for few sHsps. Among these, Hsp12 has previously been demonstrated to be involved in the pathogenesis of the fungus by our group. In the present study we further investigated the biological function of the protein in the pathogenic development of U. maydis. Analysis of the primary amino acid sequence of Hsp12 in combination with spectroscopic methods to analyse secondary protein structures revealed an intrinsically disordered nature of the protein. We also carried out detailed analysis on the protein aggregation prevention activity associated with Hsp12. Our data suggest Hsp12 has trehalose-dependent protein aggregation prevention activity. Through assaying the interaction of Hsp12 with lipid membranes in vitro we also showed the ability of U. maydis Hsp12 to induce stability in lipid vesicles. U. maydis hsp12 deletion mutants exhibited defects in the endocytosis process and delayed completion of the pathogenic life cycle. Therefore, U. maydis Hsp12 contributes to the pathogenic development of the fungus through its ability to relieve proteotoxic stress during infection as well as its membrane-stabilizing function.

Deciphering the antimicrobial, antibiofilm and membrane stabilizing synergism of Mikania scandens (L.) Willd. leaves and stems substantiation through in vitro and in silico studies.

Considering the traditional application of Mikania scandens (L.) Willd. against wounds and itching. Leaves (MSL) and stems (MSS) were sequentially extracted using solvents petroleum-ether, carbon-tetrachloride, chloroform, ethyl-acetate and ethanol. Disk-diffusion assay revealed the ethyl acetate MSL and MSS extracts were the prominent against ten bacteria, five carbapenem-resistant bacteria and one fungal strains. Subsequent quantitative antimicrobial analysis specified MSL extractives more potent over MSS with lower 1500 and 3500µg/ml MIC and MBC value in both gram-negative and positive bacteria. These sturdiest ethyl-acetate MSL extractives antimicrobial efficiency also fostered fungicidal activity having lower 100µg/ml MFC. Whereat, almost homologous 160-180 min timing noted liken to standard ciprofloxacin susceptibility in both strains, 75% biofilm inhibition at 2×MIC concentration along with 92±0.2% membrane stabilizing activities over synthetic counterparts prospected in preceding standard extractives. Computational molecular docking of MSL compounds supported this findings therefore forego this valuable synergistic insight as antimicrobial agents to efficiently eradicate human infections.

Integration of Trehalose Lipids with Dissociative Trehalose Enables Cryopreservation of Human RBCs.

Cryopreservation of red blood cells (RBCs) is imperative for transfusion therapy, while cryoprotectants are essential to protect RBCs from cryoinjury under freezing temperatures. Trehalose has been considered as a biocompatible cryoprotectant that naturally accumulates in organisms to tolerate anhydrobiosis and cryobiosis. Herein, we report a feasible protocol that enables glycerol-free cryopreservation of human RBCs by integration of the synthesized trehalose lipids and dissociative trehalose through ice tuning and membrane stabilization. Typically, in comparison with sucrose monolaurate or trehalose only, trehalose monolaurate was able to protect cell membranes against freeze stress, achieving 96.9 ± 2.0% cryosurvival after incubation and cryopreservation of human RBCs with 0.8 M trehalose. Moreover, there were slight changes in cell morphology and cell functions. It was further confirmed by isothermal titration calorimetry and osmotic fragility tests that the moderate membrane-binding activity of trehalose lipids exerted cell stabilization for high cryosurvival. The aforementioned study is likely to provide an alternative way for glycerol-free cryopreservation of human RBCs and other types of cells.

Lycopene, but not zeaxanthin, serves as a skeleton for the formation of an orthorhombic organization of intercellular lipids within the lamellae in the stratum corneum: Molecular dynamics simulations of the hydrated ceramide NS bilayer model.

Carotenoids play an important role in the protection of biomembranes against oxidative damage. Their function depends on the surroundings and the organization of the lipid membrane they are embedded in. Carotenoids are located parallel or perpendicular to the surface of the lipid bilayer. The influence of carotenoids on the organization of the lipid bilayer in the stratum corneum has not been thoroughly considered. Here, the orientation of the exemplary cutaneous carotenoids lycopene and zeaxanthin in a hydrated ceramide NS24 bilayer model and the influence of carotenoids on the lateral organization of the lipid bilayer model were studied by means of molecular dynamics simulations for 32 °C and 37 °C. The results confirm that lycopene is located parallel and zeaxanthin perpendicular to the surface of the lipid bilayer. The lycopene-loaded lipid bilayer appeared to have a strong orthorhombic organization, while zeaxanthin-loaded and pure lipid bilayers were organized in a disordered hexagonal-like and liquid-like state, respectively. The effect is stronger at 32 °C compared to 37 °C based on p-values. Therefore, it was assumed that carotenoids without hydroxyl polar groups in their structure facilitate the formation of the orthorhombic organization of lipids, which provides the skin barrier function. It was shown that the distance between carotenoid atoms matched the distance between atoms in the lipids, indicating that parallel located carotenoids without hydroxyl groups serve as a skeleton for lipid membranes inside the lamellae. The obtained results provide reasonable prediction of the overall qualitative properties of lipid model systems and show the importance of parallel-oriented carotenoids in the development and maintenance of the skin barrier function.

Cryopreservation of Semen in Domestic Animals: A Review of Current Challenges, Applications, and Prospective Strategies.

Cryopreservation is a way to preserve germplasm with applications in agriculture, biotechnology, and conservation of endangered animals. Cryopreservation has been available for over a century, yet, using current methods, only around 50% of spermatozoa retain their viability after cryopreservation. This loss is associated with damage to different sperm components including the plasma membrane, nucleus, mitochondria, proteins, mRNAs, and microRNAs. To mitigate this damage, conventional strategies use chemical additives that include classical cryoprotectants such as glycerol, as well as antioxidants, fatty acids, sugars, amino acids, and membrane stabilizers. However, clearly current protocols do not prevent all damage. This may be due to the imperfect function of antioxidants and the probable conversion of media components to more toxic forms during cryopreservation.

Achieving high intracellular trehalose in hRBCs by reversible membrane perturbation of maltopyranosides with synergistic membrane protection of macromolecular protectants.

Trehalose is considered as a biocompatible cryoprotectant for solvent-free cryopreservation of cells, but the difficulty of the current trehalose delivery platforms to human red blood cells (hRBCs) limits its wide applications. Due to cell injuries caused by incubation at 37 °C and low intracellular loading efficiency, development of novel methods to facilitate trehalose entry in hRBCs is essential. Herein, a reversible membrane perturbation and synergistic membrane stabilization system based on maltopyranosides and macromolecular protectants was constructed, demonstrating the ability of efficient trehalose loading in hRBCs at 4 °C. Results of confocal laser scanning microscopy exhibited that the intracellular loading with the assistance of maltopyranosides was a reversible process, while the membrane protective effect of macromolecular protectants on trehalose loading in hRBCs was necessary. It was suggested that introduction of 30 mM poly(vinyl pyrrolidone) 8000 combined with 1 mM dodecyl-ß-D-maltopyranoside and 0.8 M trehalose could increase the intracellular trehalose to 84.0 ± 11.3 mM in hRBCs, whereas poly(ethylene glycol), dextran, human serum albumin or hydroxyethyl starch had a weak effect. All the macromolecular protectants could promote the cryosurvival of hRBCs, exhibiting membrane stabilization, and incubation and followed by cryopreservation did not change the basic functions and normal morphology of hRBCs substantially. This study provided an alternative strategy for glycerol-free cryopreservation of cells and the delivery of membrane-impermeable cargos.

AgVI and Ag/ZnOVI nanostructures from Vateria indica (L.) exert antioxidant, antidiabetic, anti-inflammatory and cytotoxic efficacy on triple negative breast cancer cells in vitro.

Human triple-negative breast cancer (TNBC) being an aggressive cancer type accounts for about 15-20% of global breast cancer cases. In the present study, the cytotoxicity of pure silver (AgVI) and silver/zinc oxide (Ag/ZnOVI) nanostructures was evaluated against the TNBC cells. The nanostructures synthesized from a green route using Vateria indica (L.) fruit extract were characterized to scrutinize their formation, crystal phase, size, shape, and surface properties via FTIR, PXRD, FE-SEM coupled with EDS spectroscopy, and BET analysis. The results of the studies have unveiled the formation of 26.43 nm and 20.97 nm sized AgVI and Ag/ZnOVI nanostructures in their purest form. The in-vitro anticancer study performed on human TNBC cells [MDA-MB468] revealed the enhancement in the antiproliferative potentiality of bimetallic Ag/ZnOVI nanostructures from 66.99 ± 0.13 to 79.73 ± 0.23 in comparison to pure AgVI nanostructures. In addition to this, the greenish yellow-fluorescence observed in the TNBC nuclei during the AO-EB staining study manifested the early apoptosis. Furthermore, the anti-inflammatory and cytotoxicity study performed on the human RBC and normal NIH3T3 murine fibroblasts cells proved the biocompatibility and non-toxic nature of the synthesized nanostructures with membrane stabilization percentage up to 94.5 ± 0.001. Additionally, the antioxidant and antidiabetic studies carried out have corroborated the radical scavenging and α-amylase inhibition capability up to 85.87 ± 0.001 and 89.60 ± 0.002 % respectively. Thus the overall results of the study substantiate the superlative antioxidant, antidiabetic, and antiproliferative property of green synthesized AgVI and Ag/ZnOVI nanostructures with excellent biocompatibility.

A new antidiabetic and anti-inflammatory biflavonoid from Schinus polygama (Cav.) Cabrera leaves.

A new biflavonoid, luteolin-(6→8'')-apigenin was isolated from 80% methanol extract of Schinus polygama (Cav.) Cabrera leaves (Anacardiaceae). The structure was elucidated by 1D and 2D-NMR spectroscopic data. This compound exhibited in vitro antidiabetic effect via α-amylase assay. Furthermore, it possesses anti-inflammatory activity through membrane stabilization effect on erythrocytes.

Multiple Na,K-ATPase Subunits Colocalize in the Brush Border of Mouse Choroid Plexus Epithelial Cells.

(1) Background: The unusual accumulation of Na,K-ATPase complexes in the brush border membrane of choroid plexus epithelial cells have intrigued researchers for decades. However, the full range of the expressed Na,K-ATPase subunits and their relation to the microvillus cytoskeleton remains unknown. (2) Methods: RT-PCR analysis, co-immunoprecipitation, native PAGE, mass spectrometry, and differential centrifugation were combined with high-resolution immunofluorescence histochemistry, proximity ligase assays, and stimulated emission depletion (STED) microscopy on mouse choroid plexus cells or tissues in order to resolve these issues. (3) Results: The choroid plexus epithelium expresses Na,K-ATPase subunits α1, α2, ß1, ß2, ß3, and phospholemman. The α1, α2, ß1, and ß2, subunits are all localized to the brush border membrane, where they appear to form a complex. The ATPase complexes may stabilize in the brush border membrane via anchoring to microvillar actin indirectly through ankyrin-3 or directly via other co-precipitated proteins. Aquaporin 1 (AQP1) may form part of the proposed multi-protein complexes in contrast to another membrane protein, the Na-K-2Cl cotransporter 1 (NKCC1). NKCC1 expression seems necessary for full brush border membrane accumulation of the Na,K-ATPase in the choroid plexus. (4) Conclusion: A multitude of Na,K-ATPase subunits form molecular complexes in the choroid plexus brush border, which may bind to the cytoskeleton by various alternative actin binding proteins.

Chemical characterization and anti-arthritic appraisal of Monotheca buxifolia methanolic extract in Complete Freund's Adjuvant-induced arthritis in Wistar rats.

The current study was designed to evaluate the anti-oxidant and anti-arthritic potential of a traditionally used herb, Monotheca buxifolia. The M. buxifolia methanolic extract (MBME) was prepared from the aerial parts of the plant followed by chemical characterization with GC-MS. The anti-oxidant potential of the MBME was demonstrated by DPPH scavenging activity. The effects of MBME on protein denaturation and membrane stabilization were determined by inhibition of egg albumin denaturation and RBC membrane stabilization assays, respectively. The in vivo anti-arthritic potential of the MBME at 50, 100, and 150 mg/kg/day was evaluated in Complete Freund's Adjuvant-induced polyarthritis in Wistar rats treated for 21 days. Phytochemicals, such as linolenic acid methyl ester, n-hexadecanoic acid, vitamin E, α-amyrin, and ß-amyrin were detected in the GC-MS analysis. The plant extract exhibited a 55.20 ± 0.69% scavenging of free radicals at 100 µg/ml concentration. It significantly (p < 0.05) stabilized human RBC membrane (65.06 ± 0.22%) and inhibited protein denaturation (70.53 ± 0.34%) at 100 mg/ml concentration. The diclofenac sodium (DS) and MBME at 150,100, and 50 mg/kg reduced the paw edema, restored the body weight, and altered blood parameters including CRP. The MBME significantly reduced the MDA and increased the SOD, CAT, and GSH levels in liver tissue homogenate in treated rats. The serum concentration of TNF-α and PGE2 was remarkably (p < 0.01-< 0.0001) restored by the DS and MBME dose dependently. The histopathological study showed that MBME 150 mg/kg commendably restored the ankle joint inflammation, bone erosion, and cartilage damage in polyarthritic rats. It was concluded that the anti-oxidant, anti-inflammatory and anti-arthritic effects of MBME might be attributed to phenols, flavonoids, triterpenoids, vitamin E, phytol, and other fatty acids. This study showed the anti-arthritic potential of Monotheca buxifolia and thus validates its traditional claim.

Supplements of an aqueous combination of Justicia adhatoda and Ocimum tenuiflorum boost antioxidative effects and impede hyperlipidemia.

BACKGROUND: Justicia adhatoda and Ocimum tenuiflorum, locally known as bashok and tulsi, are two ethnomedicinally important herbs that have been using as dietary supplements for several therapeutic applications. This study evaluated the combined effect of both the herbs as an antioxidative and antihyperlipidemic agent. METHODS: Antihyperlipidemic effect was assessed in a high-fat diet-induced hyperlipidemic model in Wistar albino rats. The rats were treated orally with extracts of bashok (J adhatoda, 200 mg/kg bw), tulshi (O tenuiflorum, 200 mg/kg bw), and a combination of bashok and tulshi (50:50), as well as with a reference drug, atorvastatin (10 mg/kg/day), with or without high-fat diet for 14 days. The antioxidative effect was studied using established in vitro models. The studies were supported by experimentally testing the effects of the extracts on membrane stabilization and inhibition of protein denaturation. RESULTS: The results showed that the serum lipid profile was significantly decreased in the different treatment groups, with bashok having the greatest effect. Body weights, total serum protein, LDH, and relative liver and adipose tissue weights were markedly restored towards baseline values, the lowest atherogenic index being achieved with the combined extract. The combination treatment significantly enhanced total phenolic content and antioxidative capacity and greatly potentiated membrane stabilization, but inhibition of protein denaturation was not significantly affected. CONCLUSION: The data demonstrate that a combination of Justicia adhatoda and Ocimum tenuiflorum could be developed as a food supplement with antioxidative and antihyperlipidemic benefits.

Similar Yet Different-Structural and Functional Diversity among Arabidopsis thaliana LEA_4 Proteins.

The importance of intrinsically disordered late embryogenesis abundant (LEA) proteins in the tolerance to abiotic stresses involving cellular dehydration is undisputed. While structural transitions of LEA proteins in response to changes in water availability are commonly observed and several molecular functions have been suggested, a systematic, comprehensive and comparative study of possible underlying sequence-structure-function relationships is still lacking. We performed molecular dynamics (MD) simulations as well as spectroscopic and light scattering experiments to characterize six members of two distinct, lowly homologous clades of LEA_4 family proteins from Arabidopsis thaliana. We compared structural and functional characteristics to elucidate to what degree structure and function are encoded in LEA protein sequences and complemented these findings with physicochemical properties identified in a systematic bioinformatics study of the entire Arabidopsis thaliana LEA_4 family. Our results demonstrate that although the six experimentally characterized LEA_4 proteins have similar structural and functional characteristics, differences concerning their folding propensity and membrane stabilization capacity during a freeze/thaw cycle are obvious. These differences cannot be easily attributed to sequence conservation, simple physicochemical characteristics or the abundance of sequence motifs. Moreover, the folding propensity does not appear to be correlated with membrane stabilization capacity. Therefore, the refinement of LEA_4 structural and functional properties is likely encoded in specific patterns of their physicochemical characteristics.

Anti-inflammatory properties of the ethanol extract from Clerodendrum cyrtophyllum Turcz based on in vitro and in vivo studies.

ETHNOPHARMACOLOGICAL RELEVANCE: Clerodendrum cyrtophyllum Turcz, a plant belonging to the Verbenaceae family, has been used in traditional medicine for the treatment of various inflammatory diseases in many Asian countries. AIM OF THE STUDY: The study aimed to evaluate anti-inflammatory properties of the ethanol extract from Clerodendrum cyrthophyllum Turcz leaves (EE-CC) through in vitro and in vivo models. MATERIAL AND METHODS: Total phenolic and flavonoid contents in the extract were determined using colorimetric methods and HPTLC. In red blood cell membrane stabilization model, rat erythrocyte suspension was treated with crude ethanol extract at different concentrations, the hemoglobin content of the supernatant solution released by red blood hemolysis was estimated. We also evaluated the effects of the ethanol extract from this plant on the production of nitric oxide (NO), tumor necrosis factor alpha (TNF-α) in stimulated RAW 264.7 cells. In order to elucidate its anti-inflammatory molecular mechanisms, we further evaluated the effects of the EE-CC on the expression of the inflammatory genes in inflammation-induced zebrafish model by tail-cutting using qPCR analysis. RESULTS: Colorimetric methods and HPTLC revealed high phenolic and flavonoid contents in the extract. In the red blood cell membrane stabilization model, the amount of hemoglobin released by red blood hemolysis significantly decreased in the presence of EE-CC, demonstrating a strong membrane stabilizing activity. EE-CC did not cause any toxic effect on cell viability but strongly inhibited NO and TNF-ɑ release due to LPS induction. The association with EE-CC significantly reduced the expression of cox-2, pla2, c3a, il-1(il1fma), il-8 (cxcl8b.1), tnf-α, and nf-ƙb, while increased the expression of the anti-inflammatory cytokine il-10 gene in cut-tail induced inflammation of zebrafish model. CONCLUSIONS: Taken together, the results suggest that the raw ethanol extract from C. cyrtophyllum Turcz leaves presents potent anti-inflammatory activities and may be useful for the treatment of various inflammatory diseases.

Bioactivity evaluations of leaf extract fractions from young barley grass and correlation with their phytochemical profiles.

BACKGROUND: The pressed juice of Barley Grass (BG) has become very popular among people for various assumed benefits along with many testimonies of people who have been healed from various ailments such as anemia, cancer, GI problems by consuming BG. The aim of our research was to validate the claims of its medicinal values such as chemo-protective action, high anti-oxidants, RBC membrane stabilization activity, and toxicity level. METHODS: Extracts of hexane, ethyl acetate and methanol were quantitatively estimated for total phenolic contents (TPC) and total flavonoid contents (TFC). The same extracts were assessed for their antioxidative potentials with the use of DPPH free radical scavenging assay followed by determination of HRBC membrane stabilization method, Brine Shrimp Lethality Assay (BSLA) and GC-MS analysis. RESULTS: All the extracts showed high TPC and TFC along with the stronger correlation with the antioxidant activity of the extracts suggesting phenolics and flavonoids contents of the extract might be attributed to showing antioxidant activity. The methanolic and ethyl acetate extracts of the plant also showed remarkable anti-inflammatory activity where methanolic extracts had the lowest EC50. During Brine Shrimp Lethality Assay, all extracts of BG were found to be bioactive and the degree of lethality was found to be concentration dependent. The GC-MS analysis of the methanolic extract of BG revealed 23 compounds which are reported to possess different biological activities. CONCLUSION: The study reveals the strong antioxidant and RBC membrane stabilization activity of BG. The Brine Shrimp Lethality Assay found extracts to be bioactive suggesting extracts as a promising candidate for plant-derived anti-tumor compounds. Further, studies are needed to validate the data on cancer cell lines.