Chloroform Fraction Of Extract Research Articles

Endothelium-dependent vasorelaxation effects of F5 fraction of Crinum amabile chloroform extract

BackgroundVascular dysfunction can lead to many health problems including hypertension and heart disease. The complexities of vascular dysfunction and vascular disorder-related diseases have prompted the search for many new biologically active compounds in the efforts of resolving the problems. Previous studies have shown that Amaryllidaceae alkaloids exert multiple biological activities, including the vasorelaxation effect. Crinum amabile, which is a family member of Amaryllidaceae, is believed to possess a promising pharmacological activity as a vasorelaxant.MethodThe vasorelaxation activities of Crinum amabile extracts and fractions were determined using in vitro models of phenylephrine pre-contracted intact and denuded rat aortic rings. The mechanistic pathways of vasorelaxation were investigated by pre-treatment of endothelium-intact rat aortic rings with L-NG Nitro Arginine Methyl Ester (L-NAME), methylene blue (MB), indomethacin, atropine and propranolol, respectively.ResultsThe results showed that chloroform extract (CE) of Crinum amabile exhibited the highest vasorelaxation activity, and further fractionation of CE found that its F5 fraction exerted the strongest activity. An in-depth study on the mechanistic pathway revealed that the endothelium-dependent vasorelaxation induced by F5 fraction was primarily achieved through stimulation of prostaglandin (PGI2) production and partially associated with NO/cGMP activation pathway.ConclusionFindings from this study suggest that Crinum amabile can serve as a promising candidate for the discovery and development of the new vasorelaxant drug.

Anticonvulsant Activity of Bombyx batryticatus and Analysis of Bioactive Extracts Based on UHPLC-Q-TOF MS/MS and Molecular Networking

Bombyx batryticatus (BB) is an anticonvulsant animal medicine in traditional Chinese medicine (TCM) and acts on the central nervous system. This research aimed to study the anticonvulsant effects of different polarity fractions of extracts from BB and to explore the components conferring anticonvulsant activity. Materials and methods: Crude extracts of BB at 20 g/kg were divided into different polarity fractions (petroleum ether, chloroform, ethyl acetate, water) and were administered to groups of mice before injecting pentylenetetrazol (PTZ) to induce convulsions. The animals were placed in chambers, and their behaviors were recorded for 30 min following the injection. Latency time, percent of protection, convulsion, convulsion rate, and convulsion score were determined for these mice. The compounds present in the different fractions were analyzed, and those from the fraction that conferred anticonvulsant activity were identified by high-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry (UHPLC-Q-TOF MS) and molecular networking (MN). The chloroform extract fractions (B-C) clearly increased the seizure latency time and protection percentage and decreased the convulsion percentage compared to the control group. The anticonvulsant effect of other extract fractions was not significant. Our study shows that the chloroform extract fractions (B-C) of BB have a significant anticonvulsant effect. We also identified 17 compounds including lumichrome, pheophorbide A, and episyringaresinol 4'-O-beta-d-glucopyranose that were found for the first time. The results of this study may lay the groundwork for studying compounds derived from Bombyx batryticatus and their anticonvulsant effect.

In Vitro Antimalarial Activity and Phytochemical Analysis of Aerial Parts of Artemisia fragrans Willd.

Background: Malaria is one of the most momentous transmittable diseases globally. Medicinal herbs like Artemisia species might be possible sources of new, effective, and cheap antiplasmodial products, making up the leading molecules to investigate new antimalarial drugs. The Artemisia genus, which belongs to the Asteraceae family, is a widely distributed medicinal plant in Iran. Methods: In this study, the antimalarial activities of essential oil, different extracts, and vacuum liquid chromatography (VLC) fractions of A. fragrans Willd. were examined by a cell-free β-hematin formation assay. The aerial parts of A. fragrans were extracted by a Soxhlet extractor, and essential oil was obtained by a Clevenger apparatus. Then, GC-MS analysis was used to identify volatile compounds of essential oil and the 100% VLC fraction of chloroform. Results: Among the extracts, chloroform extract illustrated considerable antimalarial activity compared to the control (P < 0.001), with the IC50 value of 1.22 ± 0.05 mg/mL. Among the fractions, 100% VLC fraction of chloroform extract illustrated potent antimalarial effects compared to the control (P < 0.001). The volatile oil demonstrated moderate antimalarial effects (P < 0.001) compared with the control. Besides, GC-MS determined that sesquiterpenes in the 100% ethyl acetate fraction of the chloroform extract and oxygenated monoterpenes in the essential oil might be responsible for the potent antimalarial activity of this plant. Conclusions: The 100% ethyl acetate fraction of chloroform extract along with the essential oil of A. fragrans indicated potent and moderate activity, possibly due to sesquiterpenes and oxygenated monoterpenes, respectively.

Bauhinia acuminata L. attenuates lung cancer cell proliferation: in vitro, in vivo and in silico approaches

BackgroundThe present study aims to explore the efficacy and suitability of Bauhinia acuminata L. to attenuate lung cancer on in vitro lung cancer cell lines, in vivo benzo(a)pyrene (BaP) induced mice model and in silico molecular docking. MethodsBoth in vitro and in vivo models were used to assess the efficacy of chloroform crude extracts of B. acuminata (CEBA) leaves fractions. For in vitro cytotoxicity analysis, A549 and Vero cancer cell lines were used. The BaP treated and untreated C57BL/6 mice were grouped and the in vivo acute toxicity levels were analyzed. Ethyl acetate (EA) fraction of CEBA was orally administrated in BaP treated mice and various parameters were measured in serum, liver, lung and kidney tissues. In addition, the histological, enzymatic, immunohistochemical, p53 expression and in silico molecular docking was carried out. ResultsCEBA was found to be cytotoxic against lung cancer cell lines. The oral administration of CEBA revealed good anti-lung cancer potential in BaP induced lung cancer mice. The enzymatic and non-enzymatic antioxidants level significantly increased after the treatment of CEBA. In addition, the biochemical and histological parameters were normal in CEBA treated animals. The molecular docking showed good binding between ligands of ethyl acetate fraction and the receptors of lung. ConclusionThis study confirmed the anti-proliferative capability of ethyl acetate fraction of chloroform extract of B. acuminata on lung cancer models. These findings may represent a promising candidate for cancer therapy and used for further investigations in the clinical studies.

English

Two new compounds were isolated from the chloroform extract fraction of stems of Solenostemma argel, purified by column chromatography and elucidated by phytochemical and spectroscopic methods as Stigmasterol and β-sitosterol glucoside. Chloroform had the highest insecticide activity against the growth of the third larval instar of Tribolium castaneum used as a test insect, when it was compared with n-hexane, ethyl acetate, n-butanol and water extracts.

α-Amylase and dipeptidyl peptidase-4 (DPP-4) inhibitory effects of Melicope latifolia bark extracts and identification of bioactive constituents using in vitro and in silico approaches

Context Melicope latifolia (DC.) T. G. Hartley (Rutaceae) was reported to contain various phytochemicals including coumarins, flavonoids, and acetophenones. Objective This study investigates the antidiabetic and antioxidant effects of M. latifolia bark extracts, fractions, and isolated constituents. Materials and methods Melicope latifolia extracts (hexane, chloroform, and methanol), fractions, and isolated constituents with varying concentrations (0.078–10 mg/mL) were subjected to in vitro α-amylase and dipeptidyl peptidase-4 (DPP-4) inhibitory assay. Molecular docking was performed to study the binding mechanism of active compounds towards α-amylase and DPP-4 enzymes. The antioxidant activity of M. latifolia fractions and compounds were determined by 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging and β-carotene bleaching assays. Results Melicope latifolia chloroform extract showed the highest antidiabetic activity (α-amylase IC50: 1464.32 μg/mL; DPP-4 IC50: 221.58 μg/mL). Fractionation of chloroform extract yielded four major fractions (CF1–CF4) whereby CF3 showed the highest antidiabetic activity (α-amylase IC50: 397.68 μg/mL; DPP-4 IC50: 37.16 μg/mL) and resulted in β-sitosterol (1), halfordin (2), methyl p-coumarate (3), and protocatechuic acid (4). Isolation of compounds 2–4 from the species and their DPP-4 inhibitory were reported for the first time. Compound 2 showed the highest α-amylase (IC50: 197.53 μM) and β-carotene (88.48%) inhibition, and formed the highest number of molecular interactions with critical amino acid residues of α-amylase. The highest DPP-4 inhibition was exhibited by compound 3 (IC50: 911.44 μM). Discussion and conclusions The in vitro and in silico analyses indicated the potential of M. latifolia as an alternative source of α-amylase and DPP-4 inhibitors. Further pharmacological studies on the compounds are recommended.

In Vivo Antioxidant Activity of Different Fractions of Indigofera Barberi Against Paracetamol-induced Toxicity in Rats.

To evaluate the in vivo antioxidant activity of chloroform extract fractions of Indigofera barberi (whole plant) against paracetamolinduced toxicity in rats. For 7 days, rats were treated with different chloroform extract fractions and toxicity was induced with a single dose of paracetamol by intraperitoneal injection. The group of animals pretreated with 100 mg/kg p.o of fraction D of Indigofera barberi improved significantly in terms of hepatic superoxide dismutase (SOD), catalase and peroxidase activities, and glutathione levels compared to the control group. The hepatic SOD, catalase, peroxidase activities, and glutathione levels in the animal groups treated with paracetamol were 33.6±0.09 μ/mg protein, 5.5±0.23 μ/mg protein, 0.131±0.15 μ/mL, and 46.1±5.81 μM, respectively. Hepatic SOD, catalase, peroxidase, and glutathione in the fraction D treated group were 61.8±0.07 μ/mg protein, 10.6±0.16 μ/mg protein, 0.913±0.23 μ/mL, and 87.6±1.4 micro molar, respectively. Therefore, the present study revealed that fraction D of Indigofera barberi has significant in vivo antioxidant activity and can be used to protect tissue from oxidative stress. From the results, fraction D of Indigofera barberi at a dose of 100 mg/kg, p.o., improved the SOD, catalase and peroxidase activities, and glutathione levels significantly. Based on this study, we can conclude that fraction D of Indigofera barberi possesses in vivo antioxidant activity and can be employed in protecting tissue from oxidative stress.

Anti-Oxidant, Anti-Malarial, and Phytochemical Studies on Muscari inconstrictum Bulbs Distributed in Iran

Background: Muscari inconstrictum Rech. f. is an ornamental and bulbous species with various medicinal and biological effects. Objectives: This study was designed to evaluate the anti-oxidant and anti-malarial activities of M. inconstrictum as one of the Iranian species of Muscari genus. Additionally, preliminary phytochemical investigation of the extracts with different polarity was performed. Methods: M. inconstrictum bulbs were extracted with n-hexane, chloroform, and methanol (MeOH) by Soxhlet apparatus, in the order of their polarity. Next, vacuum liquid chromatography (VLC) was used for the fractionation of extracts. Free radical scavenging and anti-malarial activities were investigated via DPPH (2,2-diphenyl-1-picrylhydrazyl) and cell-free β-hematin formation methods. Thin layer chromatography (TLC) and gas chromatography-mass spectrometry (GC-MS) were used for the characterization of potent fractions. Results: Among different extracts of M. inconstrictum, bulbs, chloroform, and n-hexane extracts were the most potent anti-oxidant and anti-malarial extracts, respectively. Moreover, methanolic, 80% and 100% ethyl acetate/n-hexane VLC fractions of chloroform extract showed significant anti-oxidant activities, which can be related to the presence of flavonoid and coumarin structures. Furthermore, 40% ethyl acetate/n-hexane VLC fraction of n-hexane extract with saponin structures is introduced as the most potent anti-malarial part. GC-MS analysis of methanol fraction of chloroform extract, 40% ethyl acetate/n-hexane VLC fraction of n-hexane extract, and the volatile oil of plant demonstrated the presence of phenolic monoterpenoid, fatty acid derivatives, and sesquiterpenoid structures as the main ingredients, respectively. Conclusions: It seems that more studies on M. inconstrictum are necessary to focus on pure compounds and their biological activities.

Cell migration and apoptosis in human lung cancer cells by Clove (Syzygium aromaticum) dried flower buds extract

In vitro inhibition of cell migration/wound closure and cellular apoptosis was studied of four chloroform extract fractions of clove (Syzygium aromaticum L.) on human A549 and H1299 cancer cell lines. The flow cytometry analysis was carried out with A549 and H1299 cancer cell lines. The results indicated that chloroform extract fractions of clove showed inhibition of wound closure/cell migration of A549 and H1299 and induced apoptosis of H1299. This was demonstrated by calculating low wound closure percentage in cells; treated with these fractions as compared to control (70%). The morphological features of the nuclei of the cells treated with fractions indicated chromatin compression, nuclear shrinkage and apoptotic bodies formation, which pointed out towards death by apoptosis. The flow cytometer analysis of the treated cancer cells H1299 also revealed death due to the apoptosis. These results confirmed that the chloroform extract of clove bud may be used to treat lung cancer.

Biological and toxicological evaluation of aerial parts extracts of locally grown Cleome austroarabica

The current study was conducted to prepare different polarities extracts and evaluate their antimicrobial and cytotoxic activities from the aerial parts of Cleome austroarabica (C. austroarabica) which was collected from the initial campus of the University of Nizwa. The coarse powder was extracted with direct methanol by using a maceration method. The extract residue was fractionated by using various organic solvents with increasing polarity. The agar disc and brine shrimp lethality (BSL) bioassays were used to assess the antimicrobial and cytotoxic activities of the aerial extracts of C. austroarabica. Four concentrations (2000, 1000, 500, 250 µg/ml) of each extract were used to determine the antimicrobial activity against two Gram (+) bacteria: Staphylococcus aurous (S. aurous), Enterococcus faecalis (E. aurous) and two Gram (−) bacteria: Haemophilus Influenza (H. Influenza), and Escherichia coli (E. coli). Brine shrimp organisms were used to assess the cytotoxic activities of various polarities aerial extracts and the fractions of chloroform extract. The antimicrobial activity results showed that all polarities extracts at different concentrations did not give any activity against the tested bacteria. The cytotoxic activity of all polarities aerial extracts displayed activity within the value of LC50 385.25–640.25 μg/ml in the order of chloroform > hexane > hydro alcoholic > ethyl acetate > methanol extract. However, the isolated all chloroform fractions showed high LC50 activity compared to control. The further extensive study will be needed to confirm the antimicrobial and cytotoxic activities of the crude extracts and to isolate the active ingredients from the highest activity aerial extracts.

Ursane derivatives isolated from leaves of Hylocereus undatus inhibit glycation at multiple stages

The present study was designed to evaluate the therapeutic potential of bioactive compounds from chloroform extract of the leaves of Hylocereus undatus in the formation of advanced glycation end products (AGEs) in vitro. Bioactivity-guided fractionation of chloroform extract from Hylocereus undatus afforded two novel 12-ursen-type triterpenes, 3β, 16α, 23-trihydroxy-urs-12- en-28-oic acid (1) and 3β, 6β, 19α, 22α-tetrahydroxy-urs-12-en-28-oic acid (2), as well as four known triterpenes 2α, 3β, 23-tetrahydroxy-urs-11-en-28-oic acid (3), 3β-acetoxy-28-hydroxyolean-12-ene (4), 3β, 16α-dihidroxyolean-12-ene (5) and 3β-acetoxy-olean-12-ene (6). Our results revealed that triterpenes 1–3 were able to inhibit the formation of AGEs in all tested assays. The data indicated that the triterpenes had inhibitory activity at the múltiple stages of glycation and that there might be a high potential for decreasing protein oxidation and protein glycation that can enhance glycative stress in diabetic complications.

Isolation and Characterization of Bioactive Phenolic Compounds from Abies Pindrow Aerial Parts

Abies pindrow Royle (Himalayan silver fir, family Pinaceae) has a long tradition of use as anxiolytic but not investigated scientifically. Previously, antianxiety activity of chloroform extract (CE) and ethyl acetate fraction (EAF) of methanol extract of the plant have been reported at the dose of 400 and 50 mg/kg, respectively. CE and EAF were subjected to bioactivity-guided-fractionation using column chromatography with a view to isolate anxiolytic constituents. The antianxiety activity of test samples was assessed by comparing with control as well as standard drug, diazepam (2 mg/kg, p.o.) using the elevated plus maze model. Among six fractions (F1 – F6) of CE obtained after column chromatography, F2 and F3 exhibited significant antianxiety activity at doses of 45 and 40 mg/kg, respectively. Further column chromatography of F2 and F3 led to the isolation of maltol and dipalmitin. Column chromatography of EAF yielded five fractions (F1 – F5). F1 Exhibited antianxiety activity at the dose of 15 mg/kg. Column chromatography of F1 led to the isolation of gallic acid, maltol, p-coumaric acid, 4-hydroxy benzoic acid, 3,4-dihydroxy benzoic acid, shikimic acid, pinitol, hesperitin, and apigenin. Among these compounds, maltol, gallic acid, p-coumaric acid, hesperitin, and apigenin exhibited antianxiety activity at doses of 100, 10, 30, 50 and 2 mg/kg, p.o., respectively.

Alpha-bisabolol from brown macroalga Padina gymnospora mitigates biofilm formation and quorum sensing controlled virulence factor production in Serratia marcescens

Biofilm formation, quorum sensing (QS)-regulated virulence and emergence of antibiotic resistance in bacterial pathogens lead to major health problems. In this perspective, antibiofilm agents and QS inhibitors have gained much attention to treat infections caused by antibiotic-resistant pathogens. For the first time, this investigation reports the antibiofilm and QS inhibitory potential of the brown macroalga Padina gymnospora against the nosocomial pathogen Serratia marcescens. The methanolic extract of P. gymnospora inhibited biofilm formation and the production of prodigiosin and protease. Successive solvent extraction, bioassay-guided fractionation of chloroform extract and GC-MS analysis of active fractions showed the presence of alpha-bisabolol with a relative abundance of 69 %. In vitro assays with alpha-bisabolol evidenced the potent inhibition of biofilm and QS-controlled prodigiosin, protease and swarming in S. marcescens, without exerting deleterious effect on its growth and metabolic activity. The results of this study exemplify the use of P. gymnospora and alpha-bisabolol as promising alternatives to antibiotics.

Estimation of Isolated Triterpenoid – Ursolic Acid in Verbena officinalis L. Aerial Parts Using TLC Densitometry

Verbena officinalis L. (Vervain; family-Verbenaceae),a traditionally used and medicinally promising plant, has been reported to contain triterpenoids as major class of phytoconstituents. But no work has ever been carried out on V. officinalis to isolate major chemical constituent(s), and to standardize plant material on the basis of isolated constituent(s) using TLC densitometry.The chloroform extract of V. officinalisaerial parts was prepared by extracting properly identified plant in a Soxhlet apparatus.Preliminary phytochemical screening of chloroform extract showed presence of triterpenoids. Column chromatography of chloroform extract using solvent systems viz., hexane:chloroform, chloroform and chloroform:methanol in different concentrationsyielded five fractions (F1- F5). Preliminary phytochemical screening of various fractions of chloroform extract revealed presence of triterpenoids in F2 and F3 fractions. Therefore, these fractions were further subjected to column chromatography. White colored crystals were obtained in SF1 sub-fraction separated from F2and was designated as VOC-1. Structure of VOC-1 was elucidated by IR and NMRspectral studies and was characterized as ursolic acid. Further, ursolic acid was quantified in V. officinalisaerial parts by validated TLC densitometric method. The content of ursolic acid was found to be 0.1580% in V. officinalis aerial parts.The linearity range, correlation coefficient, intra-day precision, inter-day precision, LOD, LOQ and accuracy were found to be 300-1800 ng, 0.997, 1.2% CV, 1.6% CV, 40 ng/spot, 130 ng/spot and 98.27% respectively.

Anti-arthritic activity of Barleria prionitis Linn. leaves in acute and chronic models in Sprague Dawley rats

AimThe present investigation was designed to evaluate anti-arthritic potential of ethyl acetate fractions of chloroform extract from leaves of Barleria prionitis. Materials and methodsBarleria prionitis L. leaves were defatted by petroleum ether and then successive extraction was done with chloroform and methanol by the hot Soxhlet extraction method. Chloroform extract was further fractionated with solvent ethyl acetate to obtain EABP. This fraction was evaluated at two doses 125 and 250mg/kg, against formaldehyde-induced acute non immunological and Freund’s Complete Adjuvant-induced chronic immunological arthritis in rats. Arthritis assessment, paw volume, body weight, motor incoordination and nociceptive threshold were measured.Haematological assessments of red and white blood cells, erythrocyte sedimentation rate, as well as histopathological studies were also done on day 21, after animals were sacrificed. ResultsDose dependent and significant inhibition of oedema was observed in both acute as well as chronic models. The extract at dose 250mg/kg showed most potent and significant (P⩽0.05–0.01) paw oedema inhibition which is supported by the results of body weight, biochemical parameters, motor incoordination and nociceptive threshold in Freund’s Complete Adjuvant-induced arthritis model.Treatment with EABP also decreased the histopathological alterations induced by Freund’s Complete Adjuvant. ConclusionIn the present investigation, extract protects synovial membrane by improving the health status through haematinic parameters and exhibits promising anti-arthritic activity. This finding thus supports the traditional use of Barleria prionitis for rheumatism. However, further studies are needed to carry out the isolation of active constituents of the fraction responsible for the activity.

Multi-constituent synergism is responsible for anti-inflammatory effect of Azadirachta indica leaf extract

Context: Azadirachta indica A. Juss. (Meliaceaes) leaves have been used traditionally to treat swelling and rheumatism in Indian cultures.Objective: To fractionate A. indica leaf extracts using bioactivity guided manner for identification of the active anti-inflammatory principles.Materials and methods: Polarity-gradient sequential extracts (petroleum ether, chloroform, methanol, and water) of A. indica leaves were screened for their anti-inflammatory potential using the carrageenan-induced rat paw edema model (1 g/kg). The chloroform extract was sequentially fractionated to obtain n-hexane (F-1), n-hexane-chloroform (F-2), and chloroform (F-3) fractions and their inhibitory effect on rat paw edema was evaluated (500 mg/kg). Inhibitory effect of F-2 on granuloma formation, plasma interleukin (IL-1), and tumor necrosis factor (TNF-α) was assessed at the doses of 100, 200, and 400 mg/kg using the cotton pellet assay in rats. Three sub-fractions (SF-1, SF-2, and SF-3) were obtained upon chromatography of F-2, and their inhibitory effect on cyclooxygenase was assessed at 200 µg/mL concentration. The sub-fractions were subjected to gas chromatography–mass spectrometry (GC-MS).Results: All the extracts showed significant anti-inflammatory effect; however, chloroform extract was the most effective against paw edema (53.25% inhibition). The three fractions of chloroform extract showed significant effect, while F-2 being the most potent (51.02%). F-2 demonstrated dose-dependent inhibition of granuloma and cytokines. Interestingly, all the sub-fractions of F-2 inhibited COX-1 and COX-2 with almost equal potential. GC-MS revealed that chemically the sub-fractions were totally different from each other.Discussion and conclusion: Anti-inflammatory effect of A. indica is a result of cumulative and synergistic effects of diversified constituents with varying polarities that collectively exert the effect via suppression of cyclo-oxygenases and cytokines (IL-1 and TNF-α).

Anti‐proliferative and apoptotic activities of constituents of chloroform extract of Juglans regia leaves

To evaluate anti-proliferative as well as apoptotic activities of compounds identified in chloroform extract of Juglans regia leaves, on human breast and oral cancer cell lines (MCF-7 and BHY). Column chromatography, MTT assay, flowcytometry and western blotting have all been used in the study. Bioassay-guided fractionation of chloroform extract of J.regia afforded isolation of 5-hydroxy-3,7,4'-trimethoxyflavone [1], lupeol [2], daucosterol [3], 4-hydroxy-α-tetralone [4], β-sitosterol [5], 5,7- dihydroxy-3,4'-dimethoxyflavone [6] and regiolone [7]. Structures of the compounds were established on the basis of spectroscopic analyses [Nuclear magnetic resonance (NMR) and mass]. All compounds inhibited proliferation of MCF-7 (human breast adenocarcinoma) and BHY (human oral squamous carcinoma) cells in a concentration-dependent manner. Compounds 6 and 7 had potent cytotoxic effects on both MCF-7 and BHY cells (IC50 21-51μm), yet were not toxic to normal cells. MCF-7 growth inhibition was attributed to apoptosis; population of apoptotic cells increased from 1.12% in controls to 5.64 and 8.1% after 48-h treatment with compounds 6 and 7, indicating their potential at inducing early and late apoptosis. The caspase cascade was not activated, as indicated by only insignificant cleavage of caspase-3. Our results suggest that compounds 6 and 7 can induce apoptosis in MCF-7 cells through the caspase-3 independent pathway.

Anti-oxidant studies and anti-microbial effect of Origanum vulgare Linn in combination with standard antibiotics.

Background:Origanum is one of the over 200 genera in the Lamiaceae (mint family), and this genus includes culinary, fragrant, and medicinal properties. The plant is reported to contain anti-microbial properties, but it lacks combination studies with that of synthetic antibiotics.Aim:To investigate the anti-oxidant and anti-microbial interaction studies of Origanum vulgare with standard drugs against Bacillus species of bacteria and Aspergillus niger.Materials and Methods:The anti-oxidant properties of phenolic, non-phenolic fractions of chloroform extract and volatile oil were evaluated by free radical-scavenging, hydrogen peroxide radical-scavenging assay, reducing power, and metal chelating assays.Results:The minimum inhibitory concentration and fractional inhibitory concentration index were determined which demonstrates the behavior of volatile oil, phenolic, and non-phenolic fractions of volatile oil with that of ciprofloxacin and fluconazole. The IC50 value for volatile oil was found to be 15, 30, and 30 μg/ml and that of phenolic fraction was 60, 120, and 120 μg/ml for free radical-scavenging, hydrogen peroxide-scavenging, and metal chelating assays respectively. Non-phenolic fraction was found to act antagonistically along with ciprofloxacin against B. cereus and B. subtilis, while the phenolic fraction exhibited indifferent activity along with ciprofloxacin against both the bacterial strains.Conclusion:This combination of drug therapy will not only prove effective in antibiotic resistance, but these natural constituents will also help in preventing body from harmful radicals which lead to fatal diseases.

In vitro Synergistic/Antagonistic Antimicrobial Effect of Culinary Herb Oreganum vulgare with Antibiotic and Antifungal Agents

Spices and culinary herbs are used traditionally as medicines to cure health problems. Researchers have investigated the phytochemicals present in the spices and culinary herbs that are responsible for biological activities. These are potent antimicrobials and can be used to replace synthetic antioxidants and preservatives like BHA and BHT used in the food industry, the long-term usage of which can cause health hazards. In the present study, antimicrobial activity of Oreganum vulgare essential oil and phenolic and non-phenolic fractions of chloroform extract was evaluated alone and in combination with ciprofloxacin against S. aureus and with fluconazole against A. terreus to combat with the emergence of antibiotic resistance.

Combination Studies of Oreganum Vulgare Extract Fractions and Volatile Oil along with Ciprofloxacin and Fluconazole against Common Fish Pathogens.

The study is aimed at finding new antibiotic therapy for aquaculture due to potential of bacteria to develop resistance to the existing therapies. Use of large quantities of synthetic antibiotics in aquaculture thus has the potential to be detrimental to fish health, to the environment and wildlife and to human health. Antimicrobial potential of volatile oil and fractions of chloroform extract of Oreganum vulgare was evaluated alone and in the presence of standard antimicrobials against common fish pathogens by disc-diffusion, agar well assay and two fold microdilution method by nanodrop spectrophotometric method. The best results were represented by volatile oil followed by phenolic fraction by disc-diffusion, agar well and microdilution assays (Minimum inhibitory concentration). By the interaction studies, it was observed that the volatile oil and phenolic fraction were able to inhibit the pathogens at very low concentration compared to standard drugs. The fractional inhibitory concentration index (FICI) was calculated and volatile oil and phenolic fractions were found to be synergistic against Pseudomonas fluorescens and Candida albicans. The experimental data suggests the use of volatile oil and phenolic fraction in combination with standard antimicrobials to maintain healthy aquaculture with lesser adverse effects as compared to synthetic antibiotic therapy.