Membrane Stabilization Assay Research Articles

A Meretrix meretrix visceral mass derived peptide inhibits lipopolysaccharide-stimulated responses in RAW264.7 cells and adult zebrafish model

The Meretrix meretrix is abundantly present in the Indian coastal areas which can be used as an important useful bioactive source for industrial applications. The M. meretrix visceral mass (MMV) was hydrolysed with four different enzymes and verified for anti-inflammatory activity with the help of HRBC membrane stabilization (HMS) and albumin denaturation (AD) assay. Among the hydrolysates, the tryptic 6th hour hydrolysate was selected for purification using ultrafiltration and size-exclusion chromatography (SEC). Further, the purified peptide was identified to have six amino acid sequence (HKGQCC, 675.582 Da). However, to confirm the anti-inflammatory effects of the purified peptide, it was investigated for nitric oxide synthase (iNOS), pro-inflammatory cytokines production as well as cyclooxygenase-2 (COX-2) activation in lipopolysaccharide (LPS)-stimulated RAW264.7 cells and also evaluated for its functional properties. The in-vitro gastrointestinal digestion was performed on the peptide which cleaved the peptide into two i.e. MMV1 (HK, 284.1 Da) and MMV2 (GQCC, 410.1 Da). The data suggested that the MMV2 peptide have maximum activity and was found to be stable at high temperatures. The MMV2 peptide demonstrated abrupt localization throughout the adult zebrafish body and successfully downregulated the mRNA levels of inflammation-related genes in LPS-induced adult zebrafish. This study indicates that the peptide MMV2 possesses anti-inflammatory activity by suppressing the induced inflammation and can be a strong competitor against non-steroidal anti-inflammatory drugs (NSAIDs).

In vitro assessment of anti-inflammatory and anti-arthritic effects of Helicanthes elasticus (Desv.) Danser accessions collected from six different hosts.

Arthritis and other rheumatoid or non-rheumatoid joint inflammations are prevalent among people all around the world. The plant-derived drugs and other herbal therapeutic practices are widely used against these malfunctions. Helicanthes elasticus (Desv.) Danser, an endemic hemiparasite was found less exploited for these types of disorders and this plant collected from different hosts might be having differential expression towards such ailments. In order to asses this differential expression, in the present study, methanolic extract of Helicanthes elasticus collected from six different hosts were examined for HRBC membrane stabilization assay for anti-inflammatory responses and the protein denaturation method for anti-arthritic assay. Anti-inflammatory and anti-arthritic potential were observed in H. elasticus growing on different hosts. The hemiparasitic plant obtained from Hevea brasiliensis, Nerium oleander and Anacardium occidentale showed high anti-inflammatory responses whereas those from Nerium oleander had high anti-arthritic efficacy. H. elasticus showed a therapeutically significant effect on arthritis and inflammatory responses in humans and the efficacy of the curative property showed variation with respect to the host plant in which the parasite has hemiparasite found growing.

A mechanistic study of Solenostemma argel as anti-rheumatic agent in relation to its metabolite profile using UPLC/HRMS

Ethnopharmacological relevanceSolenostemma argel (Argel) is a traditional perennial edible herb that is commonly used in folkloric medicine for the treatment of rheumatic pain, inflammation, bronchitis, cold, diabetes, gastrointestinal cramps, and urinary tract infections. No previous reports traced the mechanistic activity of this plant for treatment of rheumatoid arthritis in relation to its chemical constituents. Aim of the studyThe present study was designed to substantiate the anti-arthritic potential of S. argel and identification of its secondary metabolites responsible for the action using ultra-performance liquid chromatography coupled to high resolution mass spectrometry (UPLC/HRMS). Materials and methodsThe air-dried powder of S. argel was subjected to liquid-liquid fractionation method to yield polar metabolites fraction (PMF) and nonpolar metabolites fraction (NPMF) where the metabolites that represent each fraction were identified using UPLC/HRMS. The in-vitro anti-arthritic effects of both fractions were tested using protein denaturation, membrane stabilization and proteinase inhibition assays, in addition to in-vitro enzyme inhibition assays of COXs, LOX and collagenases. Adjuvant-induced arthritis (AIA) model was also established to evaluate their anti-arthritic effects in-vivo at two doses (200 and 400 mg/kg) in compared to the standard ibuprofen (5 mg/kg). Physical changes with hind paw edema and body weight gain as well as the assessment of serum rheumatoid biomarkers, inflammatory cytokines, oxidative stress markers, and the activity of hyaluronidase and β-glucouronidase enzymes were studied. The histopathological study of ankle and knee joints and immunohistochemistry of caspase-3 and TNF-α in joint synovium were also examined. ResultsThe PMF significantly (P < 0.05) reduced paw edema, serum rheumatoid markers, pro-inflammatory mediators, degeneration enzymes of cartilage and bone, and oxidative stress biomarkers. Interestingly, flavonoid glycosides and phenolic acids dominated the polar fraction, which showed the promising anti-arthritic activity of Argel compared to the NPMF which was dominated by pregnane glycosides. ConclusionsSince arthritis is a chronic disease and there are imperative needs for a lifelong treatment with desirable pharmacological action and lower cost than the currently approved synthetic drugs having severe side effects, the PMF of Argel could be used as a potent anti-rheumatic agent.

In vitro bioactivity, antimicrobial and anti-inflammatory efficacy of modified solvent evaporation assisted Trametes versicolor extract.

In the present study, modified solvent evaporation assisted methanolic Trametes versicolor extract was evaluated for its antimicrobial and anti-inflammatory efficacy. Mushroom extract showed significantly (p < 0.05) higher total phenolic content (48.71mg/g) followed by total flavonoid content (13.13mg/g), ascorbic acid content (11.03mg/g), β-carotene content (8.34), and lycopene content (6.85). Fourier transform infrared spectroscopy revealed the functional groups of the observed bioactive compounds, while HPLC chromatogram showed significantly (p < 0.05) higher gallic acid content (45.72mg/g) as compared to rutin content (12.50mg/g). Statistically, mushroom extract and artificial antioxidant (BHA) showed a non-significant (p < 0.05) difference in terms of percentage inhibition during DPPH and N2O2 scavenging assay. During the time-kill kinetics, the mushroom extract significantly inhibited the growth of S. aureus in comparison with the growth of P.aeruginosa, K. pneumonia, and E.coli. Mushroom extract showed effective anti-inflammatory activity during membrane stabilization (33.71-73.24%) and protein denaturation (23.11-74.56%) assays.

In Vitro Toxicity, Antioxidant, Anti-Inflammatory, and Antidiabetic Potential of Sphaerostephanos unitus (L.) Holttum.

Pteridophytes have been widely used in several systems of medicine. Several reports have increasingly assessed their bioactive effects, but for Sphaerostephanos unitus (L.) Holttum, only its antibacterial potential has been assessed. In this sense, the present study was carried out to reveal the phytochemical profile and to determine the toxicity, antioxidant, antidiabetic, and anti-inflammatory potential of S. unitus. Brine shrimp lethality, 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, phosphomolybdenum assay, superoxide radical scavenging activity, 2,2-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) assay (ABTS), and in vitro α-amylase inhibitory and membrane stabilization assays were applied. S. unitus extract toxicity showed variable mortality percentages, with LC50 values ranging from 4 to 30 mg/mL. DPPH radical scavenging effects of S. unitus extracts were as follows: methanol > acetone > petroleum ether > chloroform. S. unitus acetone extract displayed the strongest phosphomolybdenum reduction (10 ± 2 mg Ascorbic Acid Equivalent/g). The studied extracts also revealed efficient, superoxide scavenging effects in a dose-dependent manner. In S. unitus, the highest ABTS radical scavenging rate was observed in the chloroform extract (3000 ± 40 µmol/g). The S. unitus anti-inflammatory effect was as follows: petroleum ether > chloroform > methanol > acetone. In S. unitus extract, the highest percentage of α-amylase activity (80%) was observed for the petroleum ether extract (25 µg/mL). Faced with these findings, further studies should be performed to isolate and identify the S. unitus compounds responsible for their antioxidant, antidiabetic and anti-inflammatory effects.

GC/MS Characterization, in-vitro Antioxidant, Anti-inflammatory and Antimicrobial Activity of Essential Oils from Pinus Plant Species from Himachal Pradesh, India

The present study was aimed to explore the chemical composition and evaluate the biological efficacy of needles and stem bark oils of three Pine plant species: Pinus roxburghii, P. wallichiana, and P. gerardiana for oxidative, inflammatory and infectious diseases. The essential oils of three Pinus plants (needles and bark) were obtained by hydrodistillation and analyzed using GC/MS. The essential oils were further assessed for antioxidant, anti-inflammatory, and anti-microbial activities in-vitro. The antimicrobial activity was assessed against four bacterial strains by using agar well diffusion test. The antioxidant activity was evaluated utilizing the DPPH free radical scavenging assay and Nitric oxide assay. The GC/MS analysis leads to the identification of compounds in essential oils of P. roxburghii, P. wallichiana, and P. gerardiana. The oils showed significant anti-oxidant activity by scavenging DPPH and nitric oxide. The oils also showed promising in-vitro anti- inflammatory activities on albumin denaturation assay and HRBC membrane stabilization assay. The oils showed significant inhibition in the growth of microorganisms: Escherichia coli, Pseudomonas aurignosa, Staphylococcus aureus, and Klebsiella pneumonia, when compared to control. The observed results back the suggestion that non-polar constituents from the essential oils of Pinus plant species have the biological potential to treat oxidative, inflammatory, and bacterial infections. This may derive rationale for the medicinal properties of Pinus oil lends credence to age-old use and holds for the production of novel drugs.

In-vitro Anti-lipoxygenase and Membrane Stabilization Activity of Crude Extracts of Euphorbia lateriflora (Schum and Thonn)

Aims: This study was aimed at evaluating the in-vitro anti-inflammatory potential of crude extracts of Euphorbia lateriflora. &#x0D; Place of Study: Department of Biochemistry, Ladoke Akintola University of Technology, Ogbomoso, Nigeria.&#x0D; Methodology: The in-vitro anti-inflammatory activity of aqueous and ethanol extracts of E. lateriflora were evaluated via anti-lipoxygenase and red cell membrane stabilization assays using spectrophotometric technique.&#x0D; Results: The aqueous and ethanol extracts demonstrated significant anti-lipoxygenase activity with the aqueous extracts (IC50: 0.34 mg/ml) and the ethanol extracts (IC50: 0.25 mg/ml), showing comparable activity with standard indomethacin (0.27 mg/ml). Contrary to what was observed in the anti-lipoxygenase study, the aqueous extract demonstrated a better membrane stabilization activity with IC50 (1.13 mg/ml) than the ethanol extracts (5.44 mg/ml). The standard diclofenac drug had an IC50 of 1.02 mg/kg. Both the aqueous and the ethanol extracts demonstrated significant anti-inflammatory activity.&#x0D; Conclusion: Observation from this study established the considerable anti-inflammatory potential of this plant by inhibiting lipoxygenase activity and stabilization the membrane of the red blood cells.

Isolation of marine crab (Charybdis natator) leg muscle peptide and its anti-inflammatory effects on macrophage cells

Abstract Marine organisms provide an intriguing number of bioactive compounds. These bioactive compounds show immunomodulatory, antioxidant, anti-inflammatory and antimicrobial activities. Crabs rank third after shrimps and lobster as seafood delicacies and provide major health benefits when consumed due to presence of proteins, vitamins and unsaturated essential fatty acids. Due to their diverse biological activities, the peptides derived from marine sources may treat various diseases such as migraine, inflammatory bowel disease, rheumatoid arthritis etc. In this study the Crab Leg (CL) muscle was isolated and further hydrolysed using trypsin, alcalase and papain to produce enzymatic hydrolysates. The results obtained from degree of hydrolysis showed that the enzymes significantly cleave the proteins. Furthermore, the HPLC analysis for amino acid compositions of CL showed the sample contained both essential and non-essential amino acids where histidine (%) was highest. In vitro assays for anti-inflammatory activity like Albumin Denaturation (AD) and HRBC Membrane Stabilization (HMS) assay helped to determine the active hour hydrolysate showing the trypsin 3rd hr hydrolysate as having highest activity in CL (90.57 ± 0.95% and 49.94 ± 1.52%). This active hour hydrolysate was purified using gel filtration chromatography and the fractions were determined using AD and HMS assay which resolved fraction Ⅰ to be more active in CL. The low molecular weight peptide sequence was found to be L-G-L-G-A-A-V-L (713.456 Da) by using LC-MS/MS. Further, cell proliferation assay and ELISA were performed for the purified peptide to test the suppression of LPS-mediated induction of COX-2 in RAW264.7 macrophage cells.

Evaluation of phytochemical and In vitro anti-inflammatory activity of leaf and fruit extracts of Casuarina equisetifolia

The plant Casuarina equisetifolia, commonly called as horse tail belonging to family Casuarinaceae is used traditionally for the treatment of infections, ulcers, cough, diarrhea etc. The plant is a rich source of tannins and flavonoids. In the current study the plant leaf, fruit aqueous and ethanolic extracts were determined for total flavonoid content followed by in vitro anti-inflammatory activity study by HRBC membrane stabilization and protein denaturation assays. The results indicate that highest amount of rutin equivalent flavonoids were present in ethanolic extract of leaf, aqueous and ethanolic extracts of fruit. Fruit extracts exhibited highest % inhibition of lysis of HRBC. Aqueous leaf and fruit extracts exhibited highest inhibition of protein denaturation, The results indicate that further in vivo studies, phytochemical isolation, characterization studies could be conducted for plant extracts

Pharmacognostical Investigation of Andrographis paniculata (Green Chiretta) and Crystallization of the Bioactive component Andrographolide

Andrographis paniculata (Family: Acanthaceae) is one the most commonly used ethno-medicinal plants in certain parts of Asia and European countries. The phytochemical analysis of the leaves of A. paniculata in aqueous, methanolic, ethanolic, hydromethanolic (1:1) and hydroethanolic (1:1) extracts revealed the presence of carbohydrates, amino acid, alkaloids, saponins, tannins, flavonoids, terpenoids, glycosides, xanthoproteins and phenols. The total phenolic, flavonoid contents and FRAP values were found to be highest in the hydromethanolic extract i.e., 0.23 ± 0.008 mg GAE/g of FWt, 0.031± 0.00 mg QE/g FWt and1.261 ± 0.03 mM FeSO4 respectively. Invitro antioxidant capacity by linear regression analysis was measured by assaying DPPH radical and H2O2 scavenging capacities. The respective IC50 values of the hydromethanolic extract of the plant were found to be 86.51 μg/ml and 298.27 μg/ml. The IC50 values for in vitro anti-inflammatory activities were evaluated by heat induced protein denaturation (IC50 diclofenac = 574.06 μg/ml, IC50 APE = 179.7 μg/ml) and RBC membrane stabilization assay (IC50 diclofenac = 337.64 μg/ml, IC50 APE = 143.07 μg/ml). The IC50 values for in vitro anti-diabetic activities were evaluated by α-amylase inhibition (IC50 acarbose = 379.71 μg/ml, IC50 APE = 328.54 μg/ml). In addition, glucose diffusion was also monitored. Antimicrobial activity of the extracts was studied against common pathogens using well diffusion method. The purification of Andrographolide was carried out using different physical separation techniques such as extraction and crystallization followed by drying.

Evaluation of the antioxidant and anti-arthritic potential of Zingiber officinale Rosc. by in vitro and in silico analysis

Rheumatoid Arthritis (RA) is an autoimmune disorder that leads to joint and bone destruction. The available anti-inflammatory chemo drugs are associated with adverse side-effects providing only temporary relief. Herbal medicine can act as an alternative source for RA treatment with improved efficacy, lesser toxicity and side effects. An attempt was made to evaluate the antioxidant and anti-arthritic potential of ginger (Zingiber officinale Rosc.). Crude ginger extracts were prepared with different organic solvents using soxhlet approach. The extracts were assessed for their anti-oxidant activity using DPPH, ABTS and nitric oxide radical inhibition assays and anti-arthritic activity through membrane stabilization assay, anti-proteinase activity and protein denaturation inhibition assays. The highest scavenging potential was exhibited by Z. officinale methanolic extract (ZOME) as assessed by DPPH assay (86.26 ± 0.97%), ABTS assay (91.04 ± 0.96%) and nitric oxide assay (86.72 ± 1.51%). Similarly, the ZOME showed 84.72 ± 1.38% of inhibition in membrane stabilization assay, 82.72 ± 1.48% in anti-proteinase activity and 81.68 ± 1.66% in protein denaturation inhibition capacity. GC-MS analysis of ZOME showed 30 different phytoconstituents. The major ginger bioactive compounds were virtually docked with novel RA targets using PyRx and the pharmacokinetic properties like ADMET properties were evaluated. Among all, 8-Gingerol showed the highest covalent interaction along with suitable binding affinity to the RA targets of COX-II (−7.2), TNF-α (−4.7), MCSF (−5.0), IL-1β (−5.7) and MMP9 (−6.7) kcal/mol. 8-Gingerol can be a better drug candidate that must be further investigated for mechanistic studies to verify its therapeutic potential in treating RA.

Antioxidant, Anti-inflammatory, and Anticoagulation Properties of Aegiceras corniculatum and Acanthus ilicifolius

Free radical production from different biological and environmental sources is due to an imbalance of natural antioxidants, which further leads to inflammation. Antioxidant metabolites are often characterized by anti-inflammatory and anticoagulation activity. Mangrove plants synthesize different classes of metabolites, including antioxidants, to minimize the devastating effect of oxidation resulting from the elevated salinity, UV, and other unique geochemical components. Accordingly, this study aimed at investigating the antioxidant, anti-inflammatory, and anticoagulation properties, as well as polyphenol content of the two selected mangrove plant species: Aegiceras corniculatum and Acanthus ilicifolius. We used the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay, reducing power, ferric reducing antioxidant power (FRAP) assay, β-carotene-linoleic acid bleaching assay (BCB), total phenolic content (TPC), total flavonoid content (TFC) and total tannin content (TTC) to determine antioxidant activity of the ethanol extract of A. corniculatum bark and leaves and A. ilicifolius leaves. Furthermore, human red blood cell (HRBC) membrane stabilization assay, lipoxygenase (LOX) inhibition assay, and prothrombin time (PT) test were performed for determining anti-inflammatory activity of the samples. A. corniculatum bark is a potent antioxidant (IC50 20.49 ± 2.14 µg/mL in DPPH assay) with anti-inflammatory (IC50 23.58 ± 1.75 µg/mL in LOX inhibition assay) and anticoagulation activity (18.19 ± 0.13 min in prothrombin time assay) compared to other extracts. All extracts were found with significant (P&lt;0.001) antioxidant, anti-inflammatory, and anticoagulation properties. Further studies on liquid chromatography-mass spectrometry (LC-MS) analysis, anti-inflammatory, and anticoagulation are recommended.

Evaluation of anti-inflammatory activity of Justicia secunda Vahl leaf extract using in vitro and in vivo inflammation models

BackgroundJusticia secunda Vahl. is a medicinal plant used in ethnomedical practice as therapy to manage inflammation. Therefore, this study was designed to evaluate the anti-inflammatory activity of methanol extract of J. secunda leaves (MEJSL) using in vitro and in vivo inflammation models.MethodsSeventy-percent MEJSL was prepared following standard procedure. In vitro anti-inflammatory assays were performed using heat-induced bovine serum albumin (BSA) denaturation and erythrocyte membrane stabilization assays. Carrageenan and formaldehyde induced inflammation in rat models were used to evaluate the anti-inflammatory activity of MEJSL in vivo. Diclofenac sodium was used as a reference drug. In addition, liver and kidney function assays and hematological analysis were carried out.ResultsData revealed that varying concentrations of MEJSL significantly (P < 0.05) inhibited heat-induced BSA denaturation and stabilized erythrocyte membrane against hypotonicity-induced hemolysis when compared with diclofenac sodium in a concentration-dependent manner. In vivo study showed that 10 mg/kg body weight (b.w.) diclofenac sodium, 100 and 300 mg/kg b.w. MEJSL suppressed carrageenan-induced paw edema at the sixth hour by 71.14%, 83.08%, and 89.05%, respectively. Furthermore, 10 mg/kg b.w. diclofenac sodium, 100 and 300 mg/kg b.w. MEJSL inhibited formaldehyde-induced paw edema by 72.53%, 74.73%, and 76.48%, respectively. Animals treated with varying doses of MEJSL had reduced plasma aspartate aminotransferase and alanine aminotransferase activities; urea and creatinine concentrations; and modulated hematological parameters when compared with the untreated control group.ConclusionsFindings from this study showed that MEJSL exhibited substantial anti-inflammatory actions in the in vitro and in vivo models. It also indicated that MEJSL anti-inflammatory mechanisms of action could be through interference with phase 2 inflammatory stressors, upregulation of cytoprotective genes, stabilization of inflammatory cell membranes and immunomodulatory activity.

Enzymatic synthesis of fatty acid esters of trehalose: Process optimization, characterization of the esters and evaluation of their bioactivities.

One-pot synthesis of caprylic, lauric and palmitic acid mono- and diesters of trehalose was catalyzed by the lipase Fermase CALB™ 10000. An optimized molar conversion of 35% of trehalose to its palmitate esters was obtained in acetone at 60°C with a trehalose:palmitic acid molar ratio of 1:5 in 4h. Trehalose fatty acid esters (THFAE) were purified by column chromatography and characterized using TLC, HPTLC, HR-MS, ATR-FTIR, and differential scanning calorimetry. THFAE were studied for their antimicrobial potential against four bacterial, and two fungal species. Trehalose monolaurate and trehalose dicaprylate demonstrated MIC of 0.45mM and 16mM against Pseudomonas aeruginosa and Escherichia coli, respectively. Trehalose monocaprylate showed the highest inhibition of biofilm forming property against Staphylococcus aureus (86.25%) at 99.2mM and trehalose dipalmitate had lowest IC50 of 13.23mM. Furthermore, their anti-inflammatory property was studied in vitro using 15-LOX inhibition assay and human red blood cell membrane stabilization assay. In the confirmatory in vivo tests using carrageenan-induced rat paw edema assay, inflammation in disease control group reached up to 63% as against 32% and 20% for trehalose dilaurate and diclofenac treated groups, respectively. THFAE can hence find potential applications in pharmaceuticals, functional foods, and nutraceuticals.

Cytotoxic and decolorizing potential of Colletotrichum gloeosporioides Penz., isolated from Salacia chinensis Linn.

Salacia chinensis Linn., (Family: Celastraceae), commonly known as Saptrangi is extensively used in the treatment of diabetes, skin diseases, amenorrhea, dysmenorrhea, gonorrhoea, rheumatism etc. In the present study, in vitro anticancer activity of ethyl acetate extract of Colletotrichum gloeosporioides, an endophytic fungus isolated from S. chinensis was tested against human pancreatic cancer (PANC-1) cells following 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay method. Ability of the fungus to decolorize carcinogenic dyes such as methylene blue and congo red was carried out in malt extract broth with an incubation period of 21 days. Human red blood corpuscles membrane stabilization assay was carried out to assess the biocompatibility of fungal extract. The presence of different functional groups and few plaucible compounds in the fungal extract were detected by Nuclear magnetic resonance (13C, 1H NMR) spectroscopy and Gas chromatography mass spectroscopy (GCMS) analysis. Ethyl acetate extract of C. gloeosporioides tested at a concentration range of 50–200 μg/ml inhibited PANC-1 cell proliferation with an IC50 value of 32.86 μg/ml suggesting the possible use of fungal extract as a potent source of anticancer agent. Colletotrichum gloeosporioides grown along with the Congo red and Methylene blue, decolorized the dyes approximately upto 65% and 80% after 21 days of incubation, respectively.

Antiangiogenic, anti-inflammatory and their antioxidant activities of Turnera subulata Sm. (Turneraceae)

Abstract Turnera subulata is a substantial medicinal plant used in folk medicine to treat various ailments. The current study was assess the total phenolic and flavonoid contents to evaluate the antioxidant and anti-inflammatory activities of the sequentially extracted T. subulata plant samples. In vitro anti-angiogenic activity was evaluated by chick chorioallantoic membrane (CAM) model for chloroform, ethyl acetate and ethanol extracts. The results obtained revealed that total phenolic content of the chloroform extract (24.13 ± 0.27 mg/g) and total flavonoid content (TFC) of the chloroform extract (22.28 ± 0.40 mg/g) were found to be suggestively higher than the other extracts. A strong antioxidant property was observed for all the six extracts. A study anti-inflammatory activity was observed in chloroform and ethanol extracts, with IC50 ranging from 79 ± 1.01 μg/mL to 81 ± 1.01 μg/mL for protein denaturation assay and from 74 ± 0.11 μg/mL to 76 ± 1.11 μg/mL for HRBC membrane stabilization assay, respectively. The chloroform and ethanol extracts have exhibited good antiangiogenic property. Eventually, these results justified that the chloroform and ethanol extracts of T. subulata with great antioxidant, anti-inflammatory and antiangiogenesis potentials could be promising candidates for the development of a cost effective, potent anticancer drug with minimal side effects.

Synthesis and Molecular Drug Validations of 1H-Benzo[d]imidazol-2-amine Derivatives: Molecular Docking and its Biological Activities

Molecular adaptation of small molecules that are targeted as therapeutic agents is a most anticipated one in drug designing and development. In the present approach, a family of substituted 1H-benzo[d]imidazol-2-amine derivatives (5a-d and 6a-e) were effectively synthesised and testified for their molecular adaptations in order to develop them as novel medications against oxidation, inflammation and inflammation associated cancer types by means of in silico and in vitro assessments. Chronic inflammation, regardless of infectious agents, plays a vital role in various cancer development. Moreover, hypoxia-inflammation-cancer are highly associated together. Hydrogen peroxide free-radical scavenging, HRBC membrane stabilization assay and cell viability test by MTT assay (macrophage) were executed to establish antioxidant, anti-inflammatory and anticancer properties of these compounds. As the prostaglandin-endoperoxide synthase 2 is highly involved in inflammation and cancer development respectively, molecular docking was executed on the corresponding X-ray crystallographic models (PDB structures).

GCMS analysis and invitro antibacterial and anti-inflammatory study on methanolic extract of Thalassiosira weissflogii

Abstract Microalgae possess huge kind of natural bioactive compounds with potent values. In the present study, analysis of invitro anti inflammatory and anti bacterial activity from Thalassiosira weissflogii and anti inflammatory activity of extract was evaluated using albumin denaturation, membrane stabilization and proteinase inhibitory assay. Apart from this the methanolic extract of diatom T. weissflogii was subjected to antibacterial activity against different organisms such as E. coli, S. aureus and B. subtilis by agar well diffusion method. T. weissflogii against E. coli, S. aureus and B. subtilis was found to be 12 mm, 19 mm and 17 mm respectively. This was compared with inhibitory zone showed by 30 μg/ml of streptomycin that exhibit maximum of 18.6 mm for E. coli, 15.2 mm for S. aureus and 26 mm for B. subtilis. We found to contain various chemical constituents such as pigment, phenolic, tocopherol. The secondary metabolites of Diatom, Thalassiosira weissflogii was analysed and characterised using various analytical techniques such as UV–vis Spectroscopy and GCMS. In the present study, methanolic crude extract of T. weissflogii was extracted and examined for in vitro anti-inflammatory and antimicrobial activity. The future work will be determination of anti inflammatory activities by in vivo models.

ANTI-INFLAMMATORY ACTIVITY OF NIGELLA SATIVA SILVER NANOPARTICLES: BIOCHEMICAL STUDY

Objective: The aim of this study is to evaluate the anti-inflammatory activity of Nigella sativa silver nanoparticles (NS AgNPs).&#x0D; Methods: Fourier transform infrared analysis was used to characterize the NS AgNPs and the extract. 2,2-diphenylpicrylhydrazyl assay was done to test the antioxidant potency of NS AgNP. Furthermore, in vitro anti-inflammatory activity of the extract and the NS AgNP was determined by red blood cell (RBC) membrane stabilization assay, protein inhibition assay, and interleukin-1 (IL-1) beta assay.&#x0D; Results: The NS AgNP exhibited dose-dependent antioxidant property. At the concentration 0.01 mg/ml 80% of radical was scavenged by NS AgNP. Inhibition of protein denaturation assay also suggests that NS AgNP shows the highest activity (70%) when compared with the standard drug aspirin (65%). RBC assay suggests that NS AgNP stabilizes the RBC membrane and prevents leaking. In the enzyme-linked immunosorbent assay method the NS AgNP showed better IL-1 beta inhibition activity when compared to aqueous extract.&#x0D; Conclusion: From the study, it was inferred that NS AgNPs are more effective when compared to the extract. These results suggest that NS AgNP can be used to treat inflammatory disorders.

Aloe barbadensis Miller peptide/polypeptide fraction alleviates inflammation through inhibition of proinflammatory cytokines and mediators in vitro and in rats with Freund’s adjuvant-induced hind paw edema

Objective: To evaluate the anti-inflammatory potential of peptide/polypeptide fraction of Aloe vera through in vitro and in vivo studies. Methods: The peptide/polypeptide fraction from Aloe vera was obtained through trichloroacetic acid precipitation. The anti-inflammatory property of the peptide/polypeptide fraction was tested by protein denaturation, membrane stabilization assays. The effect of the fraction on RAW 264.7 cell viability was examined by MTT assays. The nitric oxide level was determined through Griess reagent. TNF-α and IL-6 levels were estimated using ELISA kits. In vivo studies were carried out in male Wistar rats through injection of Freund’s adjuvant in the hind paw. Paw edema was measured through the Vernier scale and levels of alanine aminotransferase, aspartate transaminase, TNF-α, IL-6, and secretory phospholipase A2 were estimated through their respective kits after fourteen days of treatment. GraphPad Prism6 was used for analyzing the results. Results: The peptide/polypeptide extract inhibited protein denaturation with an IC50 value of (218.9±15.6) μg/mL and stabilized the membrane of red blood cells with an IC50 value of (275.9±19.1) μg/mL. The extract showed no changes in cell morphology or cytotoxicity up to the concentration of 20 μg/mL in MTT assays. The peptide/polypeptide fraction markedly reduced the levels of proinflammatory markers and mediators in both in vitro and in vivo studies. Conclusions: The results indicate that the peptide/polypeptide fraction of Aloe vera has anti-inflammatory property through inhibition of inflammatory markers and mediators responsible for NF-κB and mitogen-activated protein kinase pathways.