Proteinase Inhibitory Assay Research Articles

Assessment of in vitro Anti-inflammatory and Anti-acne-inducing Bacterial Activities of Kameng (Eclipta prostrata Linn.) Cultivated in Suphan Buri Province, Thailand

This research investigates extracts from Kameng (Eclipta prostrata Linn.), originally employed in traditional medicinal recipes, with the aim of developing an extract that can serve as an alternative to antibiotics. For this purpose, the anti-inflammatory and anti-acne-inducing bacterial activities of Kameng crude ethanol extract, grown in Suphan Buri Province were investigated, with anti-inflammatory properties assessed by the inhibition of protein degradation and proteinase inhibitory assays. Findings indicated the potential of Kameng extracts as anti-inflammatory drugs that reduce protein degradation and proteinase activities. Kameng extract concentrations ranging from 0.4 to 4.0 x 104 ppm inhibited protein degradation and proteinase activity by 14.66%-50.75% and 28.57%-72.08%, respectively. The agar disc diffusion method revealed that Kameng extracts had strong inhibitory effects against S. aureus and C. acnes, with inhibition zone diameters of 31.15±0.47 and 37.28±0.47 mm, respectively. The minimal inhibitory concentration (MIC) of Kameng extract against S. aureus and C. acnes was 5.0 mg/mL and 2.5 mg/mL while minimal bactericidal concentration (MBC) of Kameng extract against S. aureus and C. acnes was 20.0 mg/mL and 10.0 mg/mL, respectively. Results suggested the potential of Kameng extract to cure acne and skin inflammation as a herbal cosmetic product.

Utilization of herbivore defensive latex from the weed Calotropis procera L in the green synthesis of silver nanoparticles and its potential application in the control of dengue vector Aedes aegypti

Milky latex produced by few and not all the plants are the treasure chest of various bioactive compounds with defensive potential against herbivores, insects and pathogens. Mankind have utilized latices in the preparation of novel plant-based therapies for centuries. Therefore, an attempt has been made to synthesis silver nanoparticles (AgNPs) using AgNO3 as a precursor and latex from C. procera as a reducing agent. The surface plasmon absorption peak at 430 nm confirmed AgNPs formation. The sharp peak with narrow width illustrated the homogenous nature. The face centre cubic structure of AgNPs was confirmed by the XRD pattern. FT-IR analysis deduced that the bio-molecules in the latex of C. procera are responsible for the reduction of AgNO3 to AgNPs. Biosynthesised L-AgNPs possessed well pronounced antibacterial activity against P. aeruginosa and K. pneumoniae. Anti-inflammatory activity of the nanoparticle determined by membrane stabilization and proteinase inhibitory assay was found to be in dose-dependent. LC50 value of L-AgNPs was found to be 63.09 µg/mL against IV instar larvae of A. aegypti. Distinct morphological changes were observed in the mosquito larva on exposure to various concentrations of L-AgNPs.

IN VITRO STUDIES ON THE ANTI-INFLAMMATORY EFFECTS OF ESSENTIAL OIL OF Rosmarinus officinalis L. IN MACROPHAGE CELLS

The use of plant-based drugs against various human disorders have attained 
 global interest. Multicomponent essential oils are used to cure inflammation 
 related disorders. The present study was aimed to assess the anti 
 inflammatory potential of essential oil from Rosmarinus officinalis (EO 
 
 RO) using in vitro model of LPS-activated RAW macrophage cell lines. 
 
 EO-RO was hydro-distilled using Clevenger apparatus and evaluated first 
 for protein denaturation and proteinase inhibitory activities, followed by 
 assays for cyclooxygenase (COX) and lipoxygenase (LOX) activities in RAW 
 264.7 macrophage cell lines. The data was statistically analysed by one-way 
 analysis of variance. Dose-dependent inhibition was observed in protein 
 denaturation with 50 µg mL-1 EO-RO exhibiting a maximum inhibition of 
 67.77%. Similarly, proteinase inhibitory assay depicted 72.47% inhibition. 
 The COX and LOX activities showed increased inhibitions in dose-dependent 
 manner i.e. 25 to 100 µg mL-1 EO-RO displayed 58.72 and 75.33%; 28.58- 
 40.86% inhibition, respectively.

Dual Antifungal and Antiproliferative Activities of a Novel Protein Fraction from a Medicinally Important Herb Trillium govanianum Wall. ex. D. Don.

Antimicrobial resistance of microorganisms and the unwanted side effects of chemoradiation therapy in cancer are major issues in healthcare. In recent times, protein-based drugs have emerged as promising candidates due to their high specificity, less side effects, etc. In this context, the rhizome of Trillium govanianum was first explored for biologically active proteins/peptides. For this, three protein fractions namely Aqueous protein fraction (APF), Hexane-Methanol-treated aqueous protein fraction (HMAPF), and Methanol-treated aqueous protein fraction (MAPF) were prepared and evaluated for antimicrobial and antiproliferative activities. In antifungal activity, HMAPF showed the lowest MIC90 values of 1.56µg/ml against Candida parapsilosis and Candida glabrata and 3.12µg/ml against Candida albicans and Candida auris. The antifungal activity was further confirmed by a chitinase assay, a growth kinetics and a proteinase inhibitory assay. Surprisingly, none of the three protein fractions exhibited antibacterial activity against Escherichia coli and Staphylococcus aureus. Moreover, APF exhibited potent antiproliferative and antioxidant activities with IC50 values of 18µg/ml and 227µg /ml, respectively. For HMAPF, an IC50 value of 70µg/ml against the MDA-MB-231 cell line was observed. The present results demonstrate that the protein fractions, particularly HMAPF and APF, might serve as potential sources of a dual antifungal and antiproliferative protein-based drug.

Evaluation of Phytochemicals, and Pharmacological Efficiency Studies of Nyctanthes Arbor-Tristis Linn

Nyctanthes arbor-tristis is a member of the Oleaceae family with various therapeutic characteristics. Different plant components are used intraditional medicine to treat illnesses such as sciatica, chronic fever, and skin diseases. It is today regarded as a rich source of numerous uniquecompounds for creating medicines against various diseases and certain industrial products. In general, the leaves, roots, flowers, and seeds of Parijatare used to cure several ailments in various dose forms, including juice and powder. The current paper discusses the Plant Profile of N. arbor-tristisLinn., as well as the Phytochemical Screening of Extract of N. arbor-tristis Leaves, physicochemical parameters, and the Extraction Procedure andPharmacological Activities conducted on N. arbor-tristis Leaves. The study aims to analyze the antioxidant, antibacterial, and anti-inflammatory potentialof petroleum ether extract of N. arbor-tristis leaves. DPPH scavenging assays and in vitro anti‐inflammatory activity was evaluated using Albumindenaturation, Proteinase inhibitory assay and was subjected to GC-MS analysis to identify its active components. The antibacterial potential wasanalyzed using the agar well diffusion method against Gram-negative and Gram-positive bacteria. Preliminary phytochemical screening confirmed thepresence of alkaloids, flavonoids, phenols, tannins, triterpenoids, Amino acids, Carbohydrate, and the absence of alkaloids, Anthraquinone, Phenols,Saponin, and tannin. Remarkably, free radicals scavenging ability was observed in DPPH with the IC50 of 610.71±13.6μg/mL, and the anti-inflammatoryassay also observed Albumin denaturation with the IC50 of 781.93±13.6μg/mL and the proteinase inhibitory activity with the IC50 of 652.98±32.5μg/mL.In the antibacterial assay, the zone of inhibition was recorded as 20.6±1.6mm, 21.3±1.8mm, 21.3±1.8mm, and 17.9±1.5mm for the strains of Escherichiacoli, Klebsiella pneumonia, Pseudomonas spp., Staphylococcus aureus, and Bacillus spp. respectively. The highest inhibition zone was observed against E. coli(24.8±1.7 and 26.6±1.4mg/mL, respectively), and the least inhibitory was observed against L. acidophilus (19.5±1.1 and 21.7±1.7mg/mL) respectively.GC-MS profile exhibited the presence of 29 components highly accountable for its pharmaceutical activities. The results suggested the presence ofphytochemicals in Nyctanthes arbor-tristis Leaves possess antioxidant and anti-inflammatory properties in the petroleum ether extract. This informationcan be used as a starting point in the search for natural-based drugs that are effective at alleviating inflammatory symptoms and antioxidant aspects.Keywords: , , , , , ,

Valorization of Adhatoda vasica leaves: Extraction, in vitro analyses and in silico approaches.

Adhatoda vasica (also called Vasaka) is a traditional medicinal herb used traditionally for the relief of cough, asthma, nasal congestion, bronchial inflammation, upper respiratory infections, bleeding disorders, skin diseases, leprosy, tuberculosis, diabetes, allergic conditions, rheumatism, tumor, and many more diseases. The present study aims to investigate the biological activities of vasicine, a potent alkaloid from A. vasica with different biological/ pharmacological assays and in silico techniques. Vasicine showed antimicrobial activity as evidenced fromthe colony-forming unit assay. It showed antioxidant activity in ABTS scavenging assay (IC50 = 11.5 μg/ml), ferric reducing power assay (IC50 = 15 μg/ml), DPPH radical scavenging assay (IC50 = 18.2 μg/ml), hydroxyl radical scavenging assay (IC50 = 22 μg/ml), and hydrogen peroxide assay (IC50 = 27.8 μg/ml). It also showed anti-inflammatory activity in proteinase inhibitory assay (IC50 = 76 μg/ml), BSA method (IC50 = 51.7 μg/ml), egg albumin method (IC50 = 53.2 μg/ml), and lipooxygenase inhibition assay (IC50 = 76 μg/ml). Vasicine showed antidiabetic activity in α-amylase inhibition assay (IC50 = 47.6 μg/ml), α-glucosidase inhibition assay (IC50 = 49.68 μg/ml), and non-enzymatic glycosylation of hemoglobin assay. It showed antiviral activity against HIV-protease (IC50 = 38.5 μg/ml). Vasicine also showed anticancer activity against lung cancer cells (IC50 = 46.5 μg/ml) and human fibroblast cells (IC50 = 82.5 μg/ml). In silico studies revealed that similar to the native ligands, vasicine also showed a low binding energy, i.e., good binding affinity for the active binding sites and interacted with α-amylase (-6.7 kcal/mol), α-glucosidase (-7.6 kcal/mol), cyclooxygenase (-7.4 kcal/mol), epidermal growth factor receptor (-6.4 kcal/mol), lipooxygenase (-6.9 kcal/mol), and HIV-protease (-6.4 kcal/mol). The present study ascertains the potential of vasicine as a bioactive compound isolated from A. vasica having therapeutic usefulness in many human diseases.

Design, synthesis and biological evaluation of novel antipyrine based α-aminophosphonates as anti-Alzheimer and anti-inflammatory agent

Herein, a series of novel antipyrine based α-aminophosphonates derivatives were synthesized and characterized. The synthesized derivatives were subjected for in vitro cholinesterase inhibition, enzyme kinetic studies, protein denaturation assay, proteinase inhibitory assay and cell viability assay. For cholinesterase inhibition, the results inferred that the test compounds possess better AChE activity (0.46 to 6.67 µM) than BuChE (2.395 to 12.47 µM). Compound 4j inhibited both AChE and BuChE (IC50 = 0.475 ± 0.12 µM and 2.95 ± 0.16 µM, respectively), implying that it serves as a dual AChE/BuChE inhibitor. Also, kinetic studies revealed that compound 4j exhibits mixed-type inhibition against both AChE and BuChE, with Ki values of 3.003 µM and 5.750 µM, respectively. Further, protein denaturation and proteinase inhibitory assays were used to test in vitro anti-inflammatory potential. It was found that compound 4o exhibited highest activity against protein denaturation (IC50 = 42.64 ± 0.19 µM) and proteinase inhibition (IC50 = 37.57 ± 0.19 µM) when compared to diclofenac. In addition, cell viability assay revealed that active compounds possess no cytotoxicity against N2a cell and RAW 264.7 macrophages. Finally, molecular docking experiments for AChE, BuChE, and COX-2 were conducted to better understand the binding modes of active compounds. Communicated by Ramaswamy H. Sarma

Four new phenolics and antiparasitic secondary metabolites from Flacourtia rukam Zoll. & Mortizi

Phytochemical investigation of Flacourtia rukam Zoll. & Mortizi (F. rukam) leaves and bark led to the isolation and characterization of seventeen compounds of which four phenolics were not previously described; 2-[(benzoyloxy)methyl]-phenyl-O-β-xylosyl-(1→2)-β-glucopyranoside (1), 2-[(benzoyloxy)methyl]-4-hydroxyphenyl-O-β-xylosyl-(1→2)-β-D-glucopyranoside (2), 2-hydroxy-5-(2-hydroxyphenoxy)phenoxy-β-glucopyranoside (3) and biphenyl-1,1′,2,2′-tetraol (5). Interestingly, the later compound is known as a synthetic but this is the first report for its isolation from nature. Chemical structures were established using extensive analysis of spectroscopic data (1 D and 2 D NMR and HRESIMS). Biphenyl-1,1,2,2′-tetrol (5) exhibited a good activity against Trypanosoma brucei trypomastigotes with IC50= 6.66 ug/mL. Compounds 2, 5, 9, 10, 11 and 12 showed a good in-vitro anti-inflammatory activity using proteinase inhibitory assay. On the contrary, all tested compounds were inactive as antileishmanial or antimalarial.

In vitro antioxidant and anti-inflammatory activities of green cardamom essential oil and in silico molecular docking of its major bioactives

Essential oil of Elettaria cardamomum (Green cardamom) is effective in the treatment of various disease conditions. Globally, Guatemala is the leading producer of green cardamom fruits. Green-cardamom fruits from Guatemala were purchased from a Saudi Arabian market. The Guatemala cardamom essential oil (GCEO) was extracted using hydro-distillation method, and analyzed using gaschromatography/ mass-spectrometry (GC-MS). The antioxidant and anti-inflammatory activities were studied using 2,2-diphenyl-1-picrylhydrazyl-hydrate, ferric-chloride, bovine serum-albumin and proteinase inhibitory assays. GC-MS results confirmed the presence of α- terpinyl acetate and 1,8-cineole as major components. The antioxidant and anti-inflammatory activities of GCEO were found to be significant compared with those of standard ascorbic acid and ibuprofen, respectively. The molecular-docking study revealed the considerable binding potential of α-terpinyl acetate and 1,8-cineole with human-peroxiredoxin-5, tyrosine-kinases and Human-5-LOX receptors. The in-silico activity prediction analysis confirmed the antioxidant and anti-inflammatory potential, whereas the pharmacokinetics confirmed the safety of most of the GCEO compounds.

Antioxidant, Anti-inflammatory and Biosorption Properties of Starch Nanocrystals In Vitro Study: Cytotoxic and Phytotoxic Evaluation

This study aimed to investigate the antioxidant, anti-inflammatory and biosorption properties of starch nanocrystals (SNC). The characterization of synthesized SNC was done using various analytical techniques like microscopic and spectroscopic analysis. The antioxidant property was determined using DPPH (2,2-diphenyl-1-picrylhydrazyl) assay and metal ion chelating assay. SNC showed the highest scavenging activity of 70.03 ± 0.74% at 100 µg/mL concentration. Protein denaturation assay and proteinase inhibitory assay depicted the anti-inflammatory property of SNC. The results revealed that the maximum inhibition activity was found at 100 µg/mL with 72.71% inhibition. The maximum removal efficiency was found to be 83.42% at pH 2.0 with 0.15 g biosorbent. As the pH increases, biosorption capacity of SNC were reduced from 8.17 to 6.30 mg/g and the efficiency of the dye removal was decreased from 80.95 to 36.01%. The shape of synthesized SNC was spherical nanoplatelets and it shows agglomeration. The Langmuir isotherm model is best suited for the biosorption experiments with the R2 value of 0.986. SNC were subjected to cytotoxic and phytotoxic evaluation. Cell viability and phytotoxic assays proves the non-toxic nature of the SNC.

TiO2@ZnO nanocomposites decorated with gold nanoparticles: Synthesis, characterization and their antifungal, antibacterial, anti-inflammatory and anticancer activities

Nanocomposite synthesis is of great interest because of their wide variety of applications in various industries. In the current investigation, the nano-sized composites of TiO2 and ZnO (TiO2@ZnO) were prepared. The prepared TiO2@ZnO nanocomposites were modified by decorating gold nanoparticles (AuNPs) (TiO2@ZnO–Au) using a hydrothermal method. Field Emission Scanning Electron Microscope (FESEM), Transmission Electron Microscope (TEM), and Energy Dispersive X-ray spectroscopy (EDX) characterized the TiO2@ZnO – Au nanocomposites. The nanomaterial group study was carried out with FT-IR spectroscopy. Then, the synthesized TiO2@ZnO–Au nanocomposites were studied for their biological applications including antifungal, antibacterial, anti-inflammatory and anticancer activities. Interestingly, TiO2@ZnO-Au nanocomposites exhibited significant antimicrobial activity towards Escherichia- coli (MTCC 25922), Staphylococcus - aureus (MTCC 25923), and Candida- albicans (MTCC 282) by in vitro process. Besides, the nanocomposites represented remarkable anti-inflammatory activity using in vitro assays including albumin denaturation inhibitory assay, proteinase inhibitory assay, and heat induced hemolysis assay. Moreover, the prepared TiO2@ZnO–Au nanocomposites exhibited significant anticancer activity towards MCF 7 cells use an IC −50 value of 59.45 μg / mL through in vitro cytotoxicity assay using MTT assay. Future studies are required to show the effective biological activity of TiO2@ZnO–Au nanocomposites through in vitro model.

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.

Probing the binding effects of zinc and cadmium with garlic phytocystatin: Implication of the abiotic stress on garlic phytocystatin.

Abiotic stress induced by heavy metals retards the growth and development of plants. Therefore, it is essential to have an insight into the potential toxic effects of heavy metals. The present article investigates the effect of zinc and cadmium on the structure and function of garlic phytocystatin (GPhyCys). The cysteine proteinase inhibitory assay showed a reduction in the inhibitory activity upon binding with zinc and cadmium. UV-vis absorption spectroscopy revealed the complex formation of zinc and cadmium with garlic phytocystatin. Fluorescence quenching experiment confirmed the quenching of fluorophores upon binding of zinc and cadmium. Synchronous and 3-dimensional fluorescence spectroscopy suggest the alteration in the microenvironment around aromatic residues of garlic phytocystatin upon binding with the above metals. Circular dichroism showed a reduction in the alpha-helical content of native garlic phytocystatin. Scanning electron micrographs showed the morphological changes in the native garlic phytocystatin upon addition of zinc and cadmium. The observations confirmed the alteration in structure and conformation of garlic phytocystatin upon interaction with zinc and cadmium. It can be safely concluded that the high concentration of zinc and cadmium can alter the functioning of cysteine proteinase present in garlic and affects the growth and development of plants.

In-vitro assessment of the binding mechanism of oxyfluorfen (herbicide) with garlic phytocystatin: multi-spectroscopic and isothermal titration calorimetric study

Oxyfluorfen (2-chloro-1-(3-ethoxy-4-nitrophenoxy)-4-(trifluoromethyl)benzene) is a nitrophenyl ether herbicide. Phytocystatins are crucial plant proteins which regulate various physiological processes and are also responsible for maintaining protease–antiprotease balance within plants. Thus, the present article deciphers the interaction of oxyfluorfen with garlic phytocystatin (GPC) through various spectroscopic and calorimetric techniques. The cysteine proteinase inhibitory assay was done to assess the inhibitory action of GPC in the presence of oxyfluorfen. The GPC loses its inhibitory activity in the presence of oxyfluorfen. The complex formation of GPC-oxyfluorfen was shown by UV absorption spectroscopy. The intrinsic fluorescence experiment affirmed the quenching of GPC in the presence of oxyfluorfen. The Stern–Volmer quenching constant and binding constant was obtained as 6.89 × 103 M−1 and 9.72 × 103 M−1, respectively. Synchronous fluorescence showed the alteration in the microenvironment around tyrosine residues. 3D fluorescence suggested the perturbation in the polarity around aromatic residues. The isothermal titration experiment suggests that the interaction of oxyfluorfen with GPC is a thermodynamically favorable reaction. Secondary structure alteration of GPC in the presence of oxyfluorfen was studied by circular dichroism (CD). The CD result showed a reduction in the α-helical content of GPC on interaction with oxyfluorfen. Consequently, all these outcomes affirmed the formation of GPC–oxyfluorfen complex along with the structural and conformational alteration. This study identifies and signifies that the exposure of oxyfluorfen induces stress within the plant system.Communicated by Ramaswamy H. Sarma

Ameliorative effect of biofabricated ZnO nanoparticles of Trianthema portulacastrum Linn. on dermal wounds via removal of oxidative stress and inflammation.

An impediment in the process of wound healing can be attributed to reactive oxygen species and inflammation. The curative efficacy of green synthesized Trianthema portulacastrum Linn. zinc oxide nanoparticles (ZnOTP) was investigated in the present study for evaluation of their wound healing potential in rodents. Total phenolic and flavonoid content of ZnOTP was determined, and antioxidant potential was evaluated by the DPPH method. In vitro anti-inflammatory activity of ZnOTP was evaluated by membrane stabilization and albumin denaturation, along with proteinase inhibitory assays. The synthesized ZnOTP were characterized by UV-Visible spectroscopy, Fourier transform infrared (FT-IR) studies, Field Emission Scanning Electron Microscopy (FESEM) and Energy Dispersive X-ray (EDX) studies. The wound healing potential of ZnOTP was monitored by excision and incision wound models. Analyses confirmed the formation of spherical nanoparticles of 10–20 nm size along with strong signals of zinc and oxygen atoms. Significant results (p < 0.05) of wound contraction rate, epithelialization and histopathology of the healed tissues of rats confirmed the promising wound healing property of ZnOTP. In addition, inflammatory markers, biochemical estimation such as the hydroxyproline content of granulation tissue, and the profile of antioxidant enzymes also supported the wound healing potential of ZnOTP. The present study advocated the attenuation of wounds via antioxidant and anti-inflammatory activities of a green synthesized nano-ointment.

In vitro anti-inflammatory activity of mangrove plant Rhizophora mucronata Lam. (Malpighiales: Rhizophoraceae)

In the present study, analysis of in vitro inflammatory showed whole plant of Rhizophora mucronata Lam. (Malpighiales: Rhizophoraceae) can be the potent source. The data from this study showed that the R. mucronata leaf, bark and root extract could serve as an important anti-inflammatory agent. Moreover, among the three extracts, the stilt root and leaves extract showed highest anti inflammatory. In vitro anti-inflammatory activity of the selected plant extracts was evaluated using albumin denaturation, membrane stabilization and proteinase inhibitory assays. As part of the investigation on the mechanism of the anti-inflammation activity, ability of extract protein denaturation was studied. Maximum inhibition (296.26%) was observed from root extract followed by bark (259.48%) and leaf (237.62%). The extracts inhibited the heat induced hemolysis of RBCs to varying degree as show in table below. The maximum inhibition 284.17% was observed from bark extract followed by root (265.05%) and leaf (232.61%). It reveals that these phytochemical constituents are responsible to maximum protection of protein denaturation, albumin denaturation and membrane stabilization assay. The future work will be determination of anti-inflammatory and anti-arthritic activities by in vivo models.

Proteinease inhibition, membrane stabilization, antioxidant and phytochemical evaluations of leaves, seeds and calyces of four selected edible medicinal plants

AbstractThe aqueous extracts of the edible parts of four common medicinal plants (Hibiscus sabdariffa, Telfairia occidentalis, Moringa oleifera and Ocimum gratissimum) readily consumed as food or supplements in many African settings are examined for some of their acclaimed pharmacological activities and possible potential associated risks. The classes of phytochemicals present are examined qualitatively and quantitatively, while the antioxidant activities were determined using three assays which include 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2′-azinobis-3-ethylbenzothiazoline-6-sulfonate (ABTS) and hydrogen peroxide (H2O2) assays. The anti-inflammatory activities were also examined using proteinase inhibitory assays and red blood cell membrane stabilisation assays. Both the antioxidant and anti-inflammatory activities of the plants indicated that the plants have potentials for applications in foods and nutraceuticals that can be used for the management of various ailments and diseases. However, the consu...

Qualitative phytochemical screening and evaluation of anti-inflammatory, analgesic and antipyretic activities of Microcos paniculata barks and fruits

ObjectiveThe main objectives of this study were to qualitatively evaluate the profile of phytochemical constituents present in methanolic extract of Microcos paniculata bark (BME) and fruit (FME), as well as to evaluate their anti-inflammatory, analgesic and antipyretic activities. MethodsPhytochemical constituents of BME and FME were determined by different qualitative tests such as Molisch's test, Fehling's test, alkaloid test, frothing test, FeCl3 test, alkali test, Salkowski's test and Baljet test. The anti-inflammatory, analgesic and antipyretic activities of the extracts were evaluated through proteinase-inhibitory assay, xylene-induced ear edema test, cotton pellet-induced granuloma formation in mice, formalin test, acetic acid-induced writhing test, tail immersion test and Brewer's yeast-induced pyrexia in mice. ResultsM. paniculata extracts revealed the presence of carbohydrates, alkaloids, saponins, tannins, flavonoids and triterpenoids. All of the extracts showed significant (P<0.05, vs aspirin group) proteinase-inhibitory activity, whereas the highest effect elicited by plant extracts was exhibited by the BME (75.94% proteinase inhibition activity) with a half-maximal inhibitory concentration (IC50) of 61.31 μg/mL. Each extract at the doses of 200 and 400 mg/kg body weight showed significant (P<0.05, vs control) percentage inhibition of ear edema and granuloma formation. These extracts significantly (P<0.05, vs control) reduced the paw licking and abdominal writhing of mice. In addition, BME 400 mg/kg, and FME at 200 and 400 mg/kg showed significant (P<0.05, vs control) analgesic activities at 60 min in the tail immersion test. Again, the significant (P<0.05, vs control) post-treatment antipyretic activities were found by BME 200 and 400 mg/kg and FME 400 mg/kg respectively. ConclusionStudy results indicate that M. paniculata may provide a source of plant compounds with anti-inflammatory, analgesic and antipyretic activities.

A novel single WAP domain-containing protein isoform with antibacterial relevance in Litopenaeus vannamei

Single WAP domain (SWD)-containing protein is a small protein containing a whey acidic protein (WAP) domain at the C-terminal region. SWD-containing protein exhibits structural similarity to the family of serine proteinase inhibitors. As of this writing, some SWD domain-containing proteins have been identified in crustaceans, and their functions included antibacterial and anti-proteinase activities. We identified a SWD protein isoform gene in Litopenaeus vanname (Lv-SWDi). Very high sequence similarity was found between Lv-SWDi and Lv-SWD. Results of time-course analysis for the gene expression profile showed that Lv-SWDi could produce a rapid feedback and an obvious upregulation at 12 h after Vibrio injection. Endogenous Lv-SWDi protein was obviously upregulated, and the highest expression level was reached at 24 h after Vibrio injection. The purified rLv-SWDi could directly bind to Gram-positive and Gram-negative bacteria. Results of the proteinase inhibitory assay also showed that rLv-SWDi could inhibit secretory protease activity from Bacillus subtilis. Lv-SWDi is a part of an important immunity-relevant gene and may serve important functions in defense against bacteria.

Study of Antioxidant, Antibacterial and Anti-Inflammatory Activity of Cinnamon (Cinamomum Tamala), Ginger (Zingiber Officinale) and Turmeric (Curcuma Longa)

Ethanol extract of ginger, turmeric and cinnamon was assessed for its antioxidant, antimicrobial and anti-inflammatory activity. The antioxidant activity was determined by measuring FRAP (ferric reducing-antioxidant power) assay. The antibacterial efficacy was determined using paper disc method against different gram negative bacterial and sensitivity in terms of zones of inhibition of all extract were also determined. In vitro anti-inflammatory activity was evaluated using Proteinase inhibitory assay. Aspirin was used as a standard drug for the study of anti-inflammatory activity. The result shows that the ethanolic extract of the ginger and turmeric were effective against all the bacteria tested, where as the ethanolic extract of cinnamon was failure in inhibiting the growth of all bacteria tested. The ethanolic extract of ginger possessed strong antioxidant activity in FRAP method. The ethanolic extracts of ginger shows the largest antioxidant FRAP value where as the turmeric ethanolic extract showed the minimum antioxidant FRAP value which were given as 3.86 mM/100gm and 0.38 mM/100gm respectively. The FRAP value for the ethanolic cinnamon extract was found to be 0.40 mM/100gm. The ethanolic extract of ginger and turmeric also showed in vitro anti-inflammatory activity by inhibiting the proteinase activity. Proteinase activity was significantly inhibited by ginger (78.49%), turmeric (66.48%) and cinnamon (58.72%) at 800ug/ml concentration. From the result it is concluded that the ginger, turmeric and cinnamon ethanol extract showed the antioxidant and anti-inflammatory activity where as the ginger and turmeric ethanol extract exhibited the antibacterial activity.