Denaturation Method Research Articles

Phenolic Content, Antioxidant Activity and In Vitro Anti-Inflammatory and Antitumor Potential of Selected Bulgarian Propolis Samples

Background/objectives: Propolis (bee glue) is a valuable bee product widely used as a natural remedy, a cosmetic ingredient, a nutritional value enhancer and a food biopreservative. The present research aims to investigate the phenolic content, antioxidant activity and in vitro anti-inflammatory and antitumor potential of six propolis samples from three regions of Bulgaria (Vidin, Gabrovo and Lovech). Methods: the analysis of propolis phenolic compounds was determined by high-performance liquid chromatography (HPLC); the antioxidant activity of ethanolic propolis extracts was assessed by the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay and ferric-reducing antioxidant power (FRAP) assay; the in vitro anti-inflammatory activity was assessed by the inhibition of albumin denaturation method; the in vitro antitumor activity was determined in human metastatic breast cancer cell line MDA-MB-231 using 3-(4,5-Dimethyl -2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. Results: The ethanolic propolis extracts exhibited the total phenolic content from 190.4 to 317.0 mg GAE/g, total flavonoid content from 53.4 to 79.3 mg QE/g and total caffeic acid derivatives content from 5.9 to 12.1 mg CAE/g. The studied propolis extracts showed significant antioxidant capacity (between 1000.3 and 1606.0 mM TE/g determined by the DPPH assay, and between 634.1 and 1134.5 mM TE/g determined by the FRAP assay). The chemical composition analysis indicated high concentrations of caffeic acid benzyl ester, chrysin, pinocembrin and pinobanksin-3-O-propionate, predominantly in three of the propolis samples originating from Gabrovo and Lovech regions. All propolis samples demonstrated promising in vitro anti-inflammatory activity, expressed as the inhibition of thermally induced albumin denaturation by 73.59% to 78.44%, which was higher than that of the conventional anti-inflammatory drugs Aspirin (58.44%) and Prednisolone Cortico (57.34%). The propolis samples exhibited high in vitro cytotoxicity against cancer cells MDA-MB-231 with IC50 values ranging between 9.24 and 13.62 µg/mL as determined by MTT assay. Conclusions: Overall, we can suggest that the high phenolic content of Bulgarian propolis from respective areas contributes to its enhanced antioxidant, anti-inflammatory and antitumor activity. Taken together, our results support the beneficial properties of Bulgarian propolis, with potential application as a promising therapeutic agent.

Study of the Biological Activities of Aethiopinone Isolated from the Hexane Extract of Salvia argentea L. Roots and its Combinations with Reference Products

Background: In recent years, chemical and biological studies have explored medicinal plants to develop new treatments for oxidative stress, inflammation, and diabetes. Objective: This study aims to characterize for the first time the chemical composition of the hexane extract from the roots of Salvia argentea using GC and GC/MS techniques. We also isolated its major compound and evaluated its biological activities, including its antioxidant, antiinflammatory, antidiabetic, and hemolytic effects, as well as the combination of this major compound with reference drugs. Methods: Antioxidant activity was assessed using DPPH, β-carotene, and phosphomolybdenic tests. The anti-inflammatory effect was measured by the egg albumin denaturation method and protein denaturation inhibition. The antidiabetic activity was determined by evaluating the inhibition of α-amylase, with acarbose as a positive reference. The combined effect of aethiopinone with standards was also studied to reduce the minimum effective dose and minimize side effects. Results: The hexane extract of Salvia argentea is mainly composed of aethiopinone (53.3%) and ferruginol (20.5%). aethiopinone was isolated and identified using spectroscopic methods, including 1H NMR, 13C NMR, and IR. The biological activity assessment showed that hexane extract and aethiopinone had promising antioxidant, anti-inflammatory, and antidiabetic properties. In addition, synergistic effects were observed when aethiopinone was combined with positive references, resulting in a significant reduction of the required minimum inhibitory concentrations. The human erythrocyte toxicity assessment showed that hexane extract and aethiopinone induced very low hemolysis levels, reaching 24.18% and 31.13%, respectively, at high concentrations of 2000 μg/mL. Conclusion: Further research is needed to confirm their therapeutic effects and assess their potential for use in the pharmaceutical industry.

Optimization of Lipopeptide Biosurfactant Production with Anti-Inflammatory and Antimicrobial Activities

Background: Biosurfactants are very important amphiphile compounds due to their interesting advantages such as low toxicity, biodegradability and also their many biological properties. Methods: In the present study, antimicrobial and anti-imflammatory activities were evaluated to determine the biological proprieties of biosurfactant BLA 2906 produced by Alcaligenes aquatilis YGD 2906 using different assays. Screening and optimization component concentrations in the medium were investigated using PBD and SRM to increase surfactant yield in term emulsification activity value (E24%). Results: The halos of antifungal activity presented a mean value of 12.33 mm to 17.67 mm. For antibacterial activity, the diameter varied from 10.33 to 12.67 mm with a very important antiinflammatory activity using a protein denaturation method that showed a maximum inhibition of 92.79%. Conclusion: These results suggest that BLA 2906 may be used as a new therapeutic and antiinflammatory agent. The PBD selected 7 significant components out of the 14 screened. The RSM resulted in the production in terms of emulsification activity of 68.37% in the optimized medium.

Gastroprotective properties of flavonoid-rich extract of Pulicaria odora against ethanol-induced gastric ulcer in mice.

Pulicaria odora has been traditionally used in northeastern Algeria for treating gastrointestinal disorders, particularly ulcers. This study aimed to assess the gastroprotective, anti-inflammatory, and antioxidant properties of a crude hydroalcoholic extract derived from the leaves of Pulicaria odora, as well as its subsequent fractions. The gastroprotective effect was studied in an ethanol-induced ulcer model in mice. The in vitro antioxidant activity was quantified via the DPPH, and ABTS methods. In vitro anti-inflammatory activity was investigated through the human red blood cell membrane stabilization method and the bovine serum albumin denaturation method. The in vivo anti-inflammatory properties of the crude extract were assessed in mice via a carrageenan-induced acute paw edema model. The crude extract significantly inhibited gastric ulcer formation in a dose-dependent manner, achieving a reduction of up to 99.91% at a dose of 200mg/kg. Among the fractions, the aqueous fraction exhibited the most potent gastroprotective effect, with a notable dose-dependent response and high ulcer inhibition rates (95.56% at 50mg/kg). Additionally, the crude extract displayed strong in vitro and in vivo anti-inflammatory activity and a high free radical scavenging capacity; with IC50 values comparable to those of ascorbic acid. LC-ESI-MSn analysis led to the identification of nine flavonoid derivatives, which were subsequently subjected to molecular docking techniques to predict their antiulcer activity. These results suggest that both glycosylated and aglycone flavonoids derived from P. odora have the potential to inhibit gastric acid secretion by targeting the proton pump.

A simple freeze-thaw based method for efficient purification of recombinant human proinsulin from inclusion bodies.

Insulin is a pivotal peptide hormone essential for regulating glucose homeostasis. It has been known for over 100 years, but its production and purification methods are still under improvement. Escherichia coli based bacterial expression system is primarily used for insulin production. The human insulin protein expressed in bacteria usually forms inclusion bodies, complicating the purification process. Traditionally, insulin purification is a time-consuming process involving urea-based denaturation methods, and various refolding techniques, followed by extensive chromatographic methods. Here, we report an easy and efficient purification of human proinsulin involving freeze-thaw based solubilization method. The extracted proinsulin inclusion bodies are treated with different concentrations of urea, followed by a freeze-thaw based solubilization. The freezing was carried out at various temperatures, mainly -80°C, -20°C, and -196°C to determine the optimum condition for solubilization. Highest solubilization of proinsulin from the inclusion body was achieved with 0.5M urea and -20°C. Further Nickel NTA-based purification was performed, and the purified protein was characterized for disulfide mapping by high-resolution mass spectrometer (HRMS). We also performed glucose uptake assays to validate the functional properties of purified proinsulin. This freeze-thaw based mild solubilization approach is a fast and effective method for getting bioactive proinsulin, which will help further design better purification and processing strategies for insulin production.

Exploring the enhanced bioactivity and reactivity of novel Fe(III) and Co(II) complexes incorporating sulfadiazine tetradentate ligand: Structural, DFT, molecular docking, and biological applications

This current work presented the synthesis, characterization and bio-evaluation of new Fe(III) (FePSD), and Co(II) (CoPSD) complexes based on N-[4-amino-5-cyano-6-(methylthio)pyridin-2-yl]-3-oxobutanamide in pyridine afford N-(4-amino-5-cyano-6-(methylthio) pyridin-2-yl)-3-oxo-2-([4-(pyrimidin-2-ylsulfamoyl) phenyl]hydrazono) butanamide (PSD). Interestingly, the FePSD and CoPSD complexes exhibit notable stability and solubility in organic solvents, with molar conductivity values of 79.35 Ω−1 cm2 mol−1 for FePSD and 8.89 Ω−1 cm2 mol−1 for CoPSD, indicating a 1:1 electrolytic nature for FePSD and non-electrolytic behavior for CoPSD. FTIR analysis confirmed metal–ligand interactions, as shifts in key bands were observed, while UV–Vis spectra and magnetic moment measurements supported octahedral geometries for both complexes. DFT calculations provided insights into the electronic structure, revealing that CoPSD has higher reactivity and electrophilicity, while FePSD showed moderate reactivity. Thermal and mass spectral analyses further characterized their stoichiometry and stability. The antibacterial and antifungal activities were evaluated against a range of Gram-positive and Gram-negative bacteria and fungal strains. The FePSD and CoPSD complexes demonstrated significantly enhanced antimicrobial efficacy compared to the parent ligand, as evidenced by larger inhibition zones, higher activity indices, and lower minimum inhibition concentrations (MIC). CoPSD exhibited superior antibacterial and antifungal activity compared to FePSD, nearing the effectiveness of standard antibiotics and antifungal drugs. Additionally, the anti-inflammatory potential of these compounds was assessed using the egg albumin denaturation method. Both FePSD and CoPSD showed remarkable inhibition of protein denaturation, surpassing the parent ligand and even outperforming the standard drug Ibuprofen at higher concentrations. The enhancement in biological activity is attributed to the metal chelation effect, which improves cell membrane permeability and reactive oxygen species (ROS) generation. Molecular docking studies further elucidated the binding interactions of PSD, FePSD, and CoPSD with target proteins, revealing stronger hydrophobic interactions and hydrogen bonding in the metal complexes, correlating with their superior biological activity. These findings highlight the potential of FePSD and CoPSD as promising antimicrobial and anti-inflammatory agents.

Anticholinesterase and Anti-Inflammatory Activities of the Essential Oils of Sawdust and Resin-Rich Bark from Azorean Cryptomeria japonica (Cupressaceae): In Vitro and In Silico Studies.

Alzheimer's disease (AD), a multifactorial neurodegenerative disorder characterized by severe cognitive impairment, affects millions of people worldwide. However, AD therapy remains limited and mainly symptomatic-focused, with acetylcholinesterase (AChE) inhibitors being the major available drugs. Thus, AD is considered by the WHO as a disorder of public health priority. Among several strategies that have been identified to combat AD, the use of natural multi-target drug ligands (MTDLs) appears to be a promising approach. In this context, we previously found that the essential oils (EOs), obtained via hydrodistillation, from Azorean Cryptomeria japonica sawdust (CJS) and resin-rich bark (CJRRB) were able to exert antioxidant activity via different mechanisms of action. Therefore, in the present work, these EOs were screened for their (i) in vitro anti-AChE and anti-butyrylcholinesterase (BChE) activities, evaluated by a modified Ellman's assay; (ii) in vitro anti-inflammatory potential, using the albumin denaturation method; and (iii) toxicity against Artemia salina. The CJRRB-EO exhibited both anti-AChE and anti-BChE activities (IC50: 1935 and 600 µg/mL, respectively), whereas the CJS-EO only displayed anti-BChE activity, but it was 3.77-fold higher than that of the CJRRB-EO. Molecular docking suggested that α-pinene and ferruginol compounds contributed to the anti-AChE and anti-BChE activities, respectively. Moreover, the anti-inflammatory activity of the CJS-EO, the CJRRB-EO, and diclofenac was 51%, 70%, and 59% (at a concentration of only 2.21 μg/mL), respectively, with the latter two presenting comparable activity. Concerning the EOs' potential toxicity, the CJRRB-EO exhibited a lower effect than the CJS-EO (LC50: 313 and 73 µg/mL, respectively). Overall, the EOs from C. japonica biomass residues, chiefly the CJRRB-EO, displayed antioxidant, anticholinesterase, and anti-inflammatory activities in a concentration-dependent manner. These properties demonstrate that these residues may be suitable natural MTDLs for AD complementary therapy when administered through aromatherapy, or, alternatively, could serve as low-cost sources of valuable ingredients, such as α-pinene.

Exploring the anti-inflammatory potential of Vitex altissima L.f. bark - An in vitro and in silico approach

Inhibition of the inflammatory response through bark polyphenolics can help to downregulate the effects of chronic inflammation and related conditions. This study mainly investigates the polyphenolic properties and their corresponding bioactivities of Vitex altissima L.f. bark. The extracts assessed for their yield percentage, secondary metabolites, and anti-inflammatory properties of V.altissima were determined using the protein denaturation method. Through in silico modeling, it confirms the binding affinity of compounds from GC-MS analysis to the inflammation protein. The results indicated that Thalavadi-III’s V.altissima recorded the highest phenolic content on 48.23 mg GAE/g extract, while Mundathurai-I exhibited the highest flavonoid content at 25.8 mg RE/g extract. Anti-inflammation properties showed the highest percentage of inhibition in Iggalore’ V.altissima . To add more significance, the compounds from GC-MS analyses were predetermined by in silico docking analyses [benzenamine, 4,4'-[sulfonylbis(4,1-phenyleneoxy)]bis- (-6.73); hematoporphyrin (-6.22)] against inflammation protein, which confirmed

Antidiabetic potential of Lavandula stoechas aqueous extract: insights into pancreatic lipase inhibition, antioxidant activity, antiglycation at multiple stages and anti-inflammatory effects.

With the increasing global prevalence of type 2 diabetes (T2D) and obesity, there is a pressing need for novel therapeutic interventions. Lavandula stoechas, a medicinal plant traditionally used for various ailments, holds promise as a potential agent for T2D management, particularly in Morocco, where it is commonly used to treat diabetes. This study aims to evaluate the pharmacological potential of L. stoechas aqueous extract (AqLs) by assessing its lipase inhibition antioxidant and anti-inflammatory activities, identifying phenolic compounds, and examining its efficacy in reducing diabetic complications. The pharmacological potential of L. stoechas aqueous extract was investigated using in vitro assays. The inhibitory effect on pancreatic lipase, antioxidant power (FRAP), and anti-inflammatory activity (albumin denaturation method) was assessed. High-performance liquid chromatography (HPLC) analysis identified phenolic compounds. Additionally, albumin glycation was evaluated by estimating fructosamine, carbonyl groups, and amyloid β-structures to assess efficacy in mitigating diabetic complications. The extract demonstrated concentration-dependent inhibition of pancreatic lipase (IC50 = 0.132 ± 0.006mg/mL), potent antioxidant activity (IC50 = 604.99 ± 1.01μg/mL), and dose-dependent anti-inflammatory effects (IC50 = 207.01 ± 34.94mg/mL). HPLC analysis revealed phenolic compounds: naringin (38.28%), syringic acid (25.72%), and cinnamic acid (15.88%) were the most abundant, with 4-hydroxybenzoic acid, hydrated catechin, and catechin ranging from 9.60% to 5.24%, and p-coumaric acid (1.73%). Furthermore, the extract inhibited albumin glycation and fructosamine production, suggesting efficacy in mitigating diabetic complications. These findings highlight the multifaceted pharmacological potential of L. stoechas aqueous extract in T2D management, suggesting that this plant can be highly beneficial for diabetic individuals.

Evaluation of the in vitro toxicity and anti-inflammatory activity of the methanolic extract of the leaves of Pistacia lentiscus L. harvested from northwestern Algeria.

Medicinal and aromatic plants have been used for thousands of years for their therapeutic properties, to treat various ailments and maintain health. This study was carried out to test the in vitro toxicity and anti-inflammatory activity of the methanolic extract of the leaves of Pistacia lentiscus L., native to the north-western region of Algeria. Toxicity was studied in vitro by the hemolysis test and anti-hemolytic activity on human red blood cells, while anti-inflammatory activity was assessed in vitro by the protein denaturation method. The study of toxicity by hemolysis of red blood cells showed a high rate of hemolysis at a dose of 200 mg/mL, with a hemolytic percentage of around 88.64%, while a high rate of anti-hemolysis was observed at a dose of 25 mg/mL, with an inhibition percentage of around 83.17%. Evaluation of anti-inflammatory activity revealed a high percentage of inhibition of ovalbumin denaturation, reaching 63.53% at the 2000 ug/mL dose, while the lowest percentage was obtained at the 3500 ug\mL concentration with 10.16%. This extract has a toxic substance but does not exceed the toxicity threshold which achieves significant anti-inflammatory activity with depandant doses.

Synthesis of 2-Phenoxyacetic Acid Analogs for Evaluation of the Anti-inflammatory Potential and Subsequent <i>In Silico</i> Analysis

Several 2-phenoxyacetic acid derivatives and their isosteres were synthesized in moderate to high yield (57–96%) for searching newer and safer alternatives to these existing anti-inflammatory agents. The compounds were characterized by 1H-NMR, 13C-NMR and HRMS. The synthesized compounds have been subjected to in vitro anti-inflammatory activity using egg albumin denaturation and human red blood cell membrane stabilization methods. The compounds were also evaluated for their anti-inflammatory activity in the mice model. The synthesized compounds were subsequently subjected to docking analysis. The in vitro pharmacokinetic properties were analyzed by applying the pkCSM software and drug-likeness was checked by using Lipinski’s rule. Some synthesized esters and amides, like SM61, SM81, SM82, SM83, SM91 and SM101, were significantly more powerful as anti-inflammatory agents. The efficacies were correlated with their orientations in the binding pocket of the cyclooxygenase enzyme. The encouraging in vitro and in vivo activities, the optimum pharmacokinetic profile, and good drug-likeness indicated that these scaffolds could be considered for future discovery of newer nonsteroidal anti-inflammatory drugs (NSAIDs).

Synthesis of New Indazole Analogs of Curcumin as Antioxidant and Anti-inflammatory Candidates: An <i>In Vitro</i> Investigation

The development of analog curcumin compounds by modifying the structure of monocarbonyl into an analog indazole of curcumin (AIC) is recognized to have a great potential. Still, only a few reports have been available. Rarely occurring in nature, indazole molecules are typically created through chemical synthesis. Therefore, this study aimed to synthesize six new AIC compounds with a particular focus on testing in vitro antioxidant activity using the DPPH and FRAP methods, as well as anti-inflammatory activity using the protein denaturation method. The results showed that the compounds formed had high anti-inflammatory activity but low antioxidant activity. All synthesis products produced higher anti-inflammatory activity than standard diclofenac sodium and curcumin compounds. Specifically, compound 3a showed the highest anti-inflammatory activity with an IC50 = 0.548 ± 0.062 μM. Therefore, it was concluded that compound 3a has the potential to be further studied for anti-inflammatory activity.

Synthesis, Molecular Docking Studies and Biological Evaluation of Thiazolyl Hydrazone Derivatives of Chromone-3-carbaldehyde as Potent Anti-Oxidant and Anti-Inflammatory Agents.

A series of 15 thiazolyl hydrazone derivatives of chromone-3- carbaldehyde have been designed and synthesized by the cyclization of thiosemicarbazone derivatives of chromone-3-carbaldehydes with 4'-substituted-2-bromo acetophenones. All these derivatives were evaluated for antioxidant activity by their direct scavenging activity objects to reactive oxygen species such as DPPH, and nitric oxide, as well as in vitro antiinflammatory activity by a protein denaturation method. Most of these synthesized compounds have shown significant antioxidant activity, among which the compounds 5b, 5c, 5e, 5g, and 5j showed very good antioxidant activities in comparison with the standard ascorbic acid. The in vitro anti-inflammatory activity revealed that the compounds 5b, 5g, and 5h possessed significant activity compared to standard diclofenac sodium. Additionally, molecular docking studies of these molecules using ovalbumin as the protein showed remarkable interactions with its active site residues, and the results indicated that the binding mode of these compounds closely resembled that of the reference compound, diclofenac sodium. Thus, these compounds represent an attractive template for the evaluation of new antiinflammatory and antioxidant agents and might be useful for exploring new therapeutic tools.

Eco-Friendly Synthesis of Cerium Nanoparticles Using Spirulina platensis: Assessing Antibacterial and Anti-inflammatory Efficacy.

Spirulina platensis, a type of cyanobacterium (blue-green algae), is well known for its rich abundant nutritional profile and bioactive compounds, which contribute to various biological functions within the human body. The application of nanotechnology toSpirulina has the potential to further enhance its biological activity in biomedical assays. This study aimed to utilize Spirulina platensisfor the green synthesis of cerium oxide nanoparticles (CeO-NPs) and evaluate their physiochemical properties. The research will assess the antibacterial and anti-inflammatory efficacy of the synthesized nanoparticles and explore the underlying mechanisms of action. Spirulinaplatensis-mediatedcerium oxide nanoparticlesare synthesized by thegreen synthesis (titration method). The biosynthesized CeO-NPs are characterized by using techniques such as UV-visible spectroscopy (UV-Vis), Fourier transform infrared spectroscopy, X-ray diffraction spectroscopy, scanning electron microscopy (SEM), and energy-dispersive X-ray (EDX). Antibacterial activity was carried out by the agar-well diffusion method and anti-inflammatory activity was carried out by the albumin denaturation method. The green synthesis of cerium oxide nanoparticles (CeO-NPs) using Spirulina, a sustainable and eco-friendly method has potential application in antibacterial and anti-inflammatory therapies. This study focuses on the green synthesis of CeO-NPs and characterizes them by using UV-Vis, Fourier transform infrared spectroscopy (FT-IR), X-ray diffraction analysis (XRD), SEM, and EDX. The UV-vis analysis confirmed the presence of CeO-NPs at a wavelength of 320 nm. FT-IR reveals four functional groups, such as C-O, N-O, and C=Cstretches. XRD analysis showed higher crystallineand less amorphous content. SEM and EDX spectra were utilized to confirm the morphology (agglomerated square shape) and the elemental composition [Ce, O, C]in the CeO-NPs. The antibacterial activity was evaluated against multidrug-resistant (MDR) clinical strains and the anti-inflammatory activity revealed significant activity in a dose-dependent manner. This study concluded that Spirulina-mediated CeO-NPs have potential as a drug in biomedical assays. Further in vitro and in vivo analysis is required to fully confirm their viability as a potential drug.

Design, Synthesis, and Anti-Prostate Cancer Potential of 2-(4-Nitrobenzyl) Malonates In Vitro and DAL Acute Oral Toxicity Assessment In Vivo.

New 4-nitrobenzyl derivatives were designed and synthesised by nucleophilic substitution reactions of 4-nitrobenzyl bromide with malonic acid and its derivatives. The synthesised molecules were characterised using mass analysis and spectroscopic techniques and tested for their antioxidant properties using various methods, such as nitric oxide, DPPH, and hydrogen peroxide radical scavenging methods. The anti-inflammatory activities of the molecules were assessed using RBC membrane stabilisation and albumin denaturation methods. We evaluated the compounds' potential anti-prostate cancer activity using the DU145 cell line. The MTT assay determined the cell viability, indicating good anti-proliferative activity. The molecule 3 c exhibited the highest potency, with a CTC50 of 11.83 μg/mL. Molecular dynamics simulations were performed to study the stability of the ligand within the protein after docking and the resulting protein-ligand complex. The in vivo analysis of molecule 3 c in the DAL xenograft model demonstrated promising results. The increase in life span, reduction in tumor volume, and comparable effects to standard drugs are encouraging features that suggest that molecule 3 c may possess significant potential as an anti-cancer agent. The research also implies that these molecules might be potential lead compounds for developing new prostate cancer drugs.

Unveiling the therapeutic potential of Canavalia rosea leaves: Exploring antioxidant, anti-inflammatory, anti-arthritic, and cytotoxic activities through biological and molecular docking evaluation with DFT analysis

BackgroundCanavalia rosea is a common tropical seacoast flowering plant from the family of Fabaceae which is reported as bay bean and coastal jack bean; has a wide range of therapeutic and nutraceutical properties. AimThe present research aims to explore some pharmacological insights of the methanol extract C. rosea of leaves (MECR) and its chloroform fraction (CFCR) and n-hexane fraction (NFCR) through in-vitro and in-silico approaches. MethodsDifferent fractions of C. rosea were subjected to ferric reduction assay and total phenolic and flavonoid content assay to explore their antioxidant potential. The anti-inflammatory activity was assessed on the hypotonic-induced human erythrocyte-lysis model, while the protein denaturation method was applied for screening anti-arthritis properties of plant extract; and the cytotoxic activity was evaluated by brine shrimp lethality bioassay. In the in-silico study, molecular docking, pass prediction and ADME/T, analysis was used to investigate anti-arthritic, anti-inflammatory, antioxidant and cytotoxic potency of five selected compounds of C. rosea. Finally, the quantum chemical density functional test analysis was applied to investigate the chemical and physical properties of those compounds. ResultsAll the soluble organic extracts of C. rosea demonstrated moderate toxicity with strong antioxidants potential, in which MECR manifested the peak level of scavenging activity (2.49) on ferric reduction assay. MECR, CFCR & NFCR significantly protected lysis of human erythrocyte membrane induced by hypotonic solution, whereas MECR and CFCR were exhibited partially equal inhibitory activity. The in-vitro anti-arthritis assay, MECR, NFCR and CFCR showed strongly significant (p˂0.001) inhibitory potency at 500 μg/mL & 1000 μg/mL concentrations. The molecular docking simulations revealed strong binding of all compounds to specific receptors, with Rutin showing the highest biological reactivity followed by Daucosterol, β-Sitosterol, Stigmasterol, and Guanidine. All the compounds showed favorable reactivity patterns, with O and H atoms poised for nucleophilic and electrophilic attacks in chemical density functional calculations. ConclusionThe recent investigation suggests that C. rosea could have the potential source of antioxidant, anti-inflammatory, anti-arthritis and cytotoxic activity, prompting further investigation into their mechanisms.

Unraveling the Potential of Mononuclear Zn(II) and Cu(II) Schiff Base Metal Complexes: Microwave‐Aided Synthesis, Theoretical Analysis and Application in Biomimetic Catalysis, CT‐DNA Interaction, and In Vitro Biological Assays

ABSTRACTGiven the widespread and diverse effects of transition metal ions such as copper and zinc in multiple biological processes, this research endeavor addresses herein the microwave‐aided synthesis of two novel tetradentate ONNO (L1) and tridentate NNO (L2) donor Schiff base ligands derived from 4,5‐dimethyl‐1,2‐phenylenediamine and 4‐bromo‐2‐hydroxybenzaldehyde in a 1:2 ratio and from 4,5‐dimethyl‐1,2‐phenylenediamine and 5‐methoxy‐2‐hydroxybenzaldehyde in a 1:1 ratio, with their respective zinc, ZnL1(1) and ZnL2(3), and copper, CuL1(2) and CuL2(4), Schiff base metal complexes, which are comprehensively characterized utilizing physiochemical and analytical techniques, along with geometry optimization using the DFT approach. Spectrophotometric analyses have been used to assess the biomimetic activity in the conversion of 3,5‐DTBC to 3,5‐DTBQ with turnover numbers 53.4 and 78 h−1 in methanol for 2 and 4, whereas the zinc complexes are found to be ineffective. With CT‐DNA, possible modes of interaction with the synthesized complexes have been explored using absorption spectroscopy, where the associated binding constants ranged between 1.541 × 105 and 2.186 × 105 M−1. EtBr‐bound DNA was used in the fluorescence quenching experiments with Stern–Volmer constant (Ksv) values in the range 4.084 × 104 and 3.402 × 104 M−1, showing a stronger association with CT‐DNA. The compounds' ability to inhibit bacterial strains was also examined in vitro, as well as their ability to reduce inflammation, through the protein denaturation method and the DPPH assay for antioxidant activity have been performed. Furthermore, molecular docking simulations have been carried out to obtain a deeper comprehension of the molecular‐level interactions with CT‐DNA and cyclooxygenase‐2 (COX‐2).

Chemical characterization and biotechnological potential of Salvia rosmarinus Spenn essential oil nanoemulsions

This study aimed to determine the total phenolic compounds, evaluate the antioxidant and antiinflammatory activities of nanoemulsions (O/W) and essential oils (EOs) from Salvia rosmarinus (rosemary). The plant material was obtained in the city of São Luís (MA). The EO was obtained by the hydrodistillation technique in a modified Clevenger extractor, and the NOE’s by phase inversion. The chemical constituents of EO were determined by GC-MS. The determination of total phenolic compounds (TPC) was performed by the Folin-Ciocalteu method. The anti-inflammatory activity was performed by the method of protein denaturation, and the antioxidant activity was performed by the spectrophotometric method of scavenging hydroxyl radicals. The GC-MS allowed quantifying 1,8-cineol (30.22%), α-pinene (22.14%), camphor (18.33%), and camphene (10.36%) as major components of the EO. The TPC of the EO was quantified at 26.74 mg EAT g-1 and the refractive index at 1.466 nD 25°. In the antioxidant activity test, an EC50 of 80.33 mgL-1 was obtained for the EO and from 19.56 to 408.85 mg L-1 for the nanoemulsions. In the anti-inflammatory activity assay, an EC50 of 62.46 mgL-1 was obtained for EO and 64.96 to 4220.25 mg L-1 for NOE’s. Finally, the pharmacological activities tested showed efficient values for EC50, therefore being considered active. This activity is attributed to the chemical compounds present, thus encouraging studies with this species aiming at its potential application in a formulated bioproduct.

Phytochemical investigation and in vitro and in vivo pharmacological activities of methanol extract of whole plant Argyreia capitiformis (Poir.) Ooststr

BackgroundA. capitiformis (Poir.) Ooststr has a long history of usage as a medicinal cure for a wide variety of illnesses in many different cultures. Pharmacological properties and phytochemical characterization of the crude A. capitiformis whole plant are evalutted, in this paper.MethodsAntioxidant activity was tested by the DPPH free radical scavenging method. In vitro anti-arthritic, anti-inflammatory, and cytotoxic effects were assessed using Bovine serum albumin (BSA), protein denaturation method, and brine shrimp mortality assays, respectively with antihelmintic activity through Pheretima Posthuma worms. Acetic acid-induced writhing, hot plate and tail immersion testing assessed in vivo analgesia. CNS activity was evaluated through elevated plaze maize, open field, hole cross, and head dipping method.ResultsPhytochemiical investigation of A. capitiformis showed the presence of alkaloid, saponin, terpennoids, steroid and flavonoids etc. with the % yield of crude 2.04%.With an IC50 of 45.35 µg/ml, the whole plant methanolic preparation has antioxidant activity equivalent to ascorbic acid. Anti-arthritic protein blocking dropped from 74.25 ± 0.12% to 12.18 ± 0.12%. 1000 µg/ml extract demonstrated 54.05 ± 0.12*% anti-inflammatory activity with protein denaturation. In the cytotoxicity assay, the extract had 129.72 µg/ml LC50 and the positive group 34.67 µg/ml. Unlike Albendazole, the methanol extract triggered mature earthworms at 50 mg/ml. The extract’s analgesic efficacy at 200 and 400 mg/kg was statistically significant (p < 0.001) in the acetic acid writhing and tail immersion method. The hot plate technique yielded statistically significant results only at 400 mg/kg (p < 0.001). Only 400 mg/kg was statistically significant in the Elevated Plaze Maize and Hole Board Procedure (p < 0.01). The hole cross and open field methods yielded highly statistically significant outcomes at 200 and 400 mg/kg (p < 0.001).ConclusionIn this research, the whole crude methanol extract of A. capitiformis revealed phytochemicals, antioxidants, in vitro anti-inflammatory and anti-arthritic properties, cytotoxicity, anti-helminthic, in vivo analgesic, and CNS inhibitory activities.

Anti-Inflammatory and Antimicrobial Activity of Silver Nanoparticles Green-Synthesized Using Extracts of Different Plants.

In this study, an easy, efficient, economical, and eco-friendly green bio-synthesis method was utilized to synthesize silver nanoparticles (AgNPs) using the extracts of four plants: Ginkgo biloba, Cichorium Intybus, Adiantum Capillus-Veneris, and Rosmarinus Officinalis. The synthesis of AgNPs was confirmed by using a uv-vis spectrometer, which showed distinct surface plasmon resonance (SPR) bands. The surface of AgNPs was characterized using scanning electron microscopy and Fourier-transform infrared spectroscopy. The anti-inflammatory activity of Tenoxicam/Meloxicam-loaded AgNPs has been studied using the inhibition of albumin denaturation method. Tenoxicam-loaded AgNPs showed higher % Inhibition, but Meloxicam-loaded AgNPs showed lower % Inhibition. Furthermore, the AgNPs showed excellent antimicrobial activity on both Gram-negative and Gram-positive bacteria.