Combinatorial effect of diclofenac with piperine and D-limonene on inducing apoptosis and cell cycle arrest of breast cancer cells

Objective:To investigate the potential synergistic activity of diclofenac with piperine and D-limonene in inducing apoptosis and cell cycle arrest in breast cancer MCF-7 cells.Methods:Molecular docking study was conducted to evaluate the binding affinity of diclofenac with piperine and D-limonene against p53,Bax,and Bcl-2.The MTT assay was used to determine IC50,and the Chou-Talay method was used to determine the synergistic concentration of the combination treatment of diclofenac plus piperine and diclofenac plus D-limonene.Apoptosis detection,cell cycle arrest,reactive oxygen species production,and mitochondrial membrane potential were also investigated.Results:Diclofenac,piperine,and D-limonene showed potent binding affinity for p53,Bax,and Bcl-2.Diclofenac plus piperine and diclofenac plus D-limonene enhanced the formation of reactive oxygen species,which also had an effect on the mitochondrial membrane’s integrity and caused DNA fragmentation.Diclofenac plus piperine and diclofenac plus D-limonene arrested the cells in the sub-G0phase while drastically lowering the percentage of cells in the G2/M phase.Furthermore,the elevated apoptosis in the combined therapy was confirmed by annexin V/propidium iodide staining.Conclusions:The combined therapy prominently enhanced the antiproliferative and apoptotic effects on MCF-7 cells compared with treatment with diclofenac,piperine,and D-limonene alone.

Survivability of freeze-dried probiotic Pediococcus pentosaceus strains GS4,GS17 and Lactobacillus gasseri(ATCC 19992)during storage with commonly used pharmaceutical excipients within a period of 120 days

Objective: To examine the survivability and stability of probiotic strains in presence and absence of pharmaceutical excipients for a long period of time at(4 ± 1)℃.Methods: The survival rates of probiotic strains, Pediococcus pentosaceus GS4(MTCC12683)(NCBI HM044322), GS17(NCBI KJ608061) and Lactobacillus gasseri(ATCC 19992), were evaluated. Probiotic strains were lyophilized individually and in combination of excipients(sorbitol, ascorbic acid, fructose and skim milk). The preparation was monitored for 120 d storing at(4 ± 1)℃. During storage, all the preparations were evaluated for viability and stability of probiotic properties like lactic acid production, antimicrobial effect, water activity, and adherence to epithelial cells.Results: Sorbitol, ascorbic acid and skim milk favoured the viability of freeze-dried cells and sustained probiotic properties during storage. Without excipients(control group),strains showed percentage of survivability not more than 70% while strains with excipients survived for 73%–93% for a long period of time.Conclusions: Commonly used excipients can be considered as a vehicle for delivering active principle in probiotic formulation and for sustaining the viability and stability of probiotic strains for a period of 120 d.

Efficacy of boswellic acid on lysosomal acid hydrolases,lipid peroxidation and anti-oxidant status in gouty arthritic mice

Objective:To evaluate the efficacy of boswellic acid against monosodium urate crystal-induced inflammation in mice.Methods:The mice were divided into four experimental groups.GroupⅠserved as control;mice in groupⅡwere injected with monosodium urate crystal;groupⅢconsisted of monosodium urate crystal-induced mice who were treated with boswellic acid(30mg/kg/b.w.);groupⅣcomprised monosodium urate crystal-induced mice who were treated with indomethacin(3mg/kg/b.w.).Paw volume and levels/activities of lysosomal enzymes,lipid peroxidation,anti-oxidant status and inflammatory mediator TNF-αwere determined in control and monosodium urate crystal-induced mice.In addition,the levels ofβ-glucuronidase and lactate dehydrogenase were also measured in monosodium urate crystal-incubated polymorphonuclear leucocytes(PMNL)in vitro.Results:The activities of lysosomal enzymes,lipid peroxidation,and tumour necrosis factor-αlevels and paw volume were increased significantly in monosodium urate crystal-induced mice,whereas the activities of antioxidant status were in turn decreased.However,these changes were modulated to near normal levels upon boswellic acid administration.In vitro,boswellic acid reduced the level ofβ-glucuronidase and lactate dehydrogenase in monosodium urate crystal-incubated PMNL in concentration dependent manner when compared with control cells.Conclusions:The results obtained in this study further strengthen the anti-inflammatory/antiarthritic effect of boswellic acid,which was already well established by several investigators.

Acaricidal activity of synthesized titanium dioxide nanoparticles using Calotropis gigantea against Rhipicephalus microplus and Haemaphysalis bispinosa

Objective:To assess the acaricidal activity of titanium dioxide nanoparticles(TiO_2 NPs)synthesized from flower aqueous extract of Calotropis gigantea(C.gigantea)against the larvae of Rhipicephalus(Boophilus)microplus[R.(B.)microplus]and the adult of Haemaphrysalis bispinosa(H.bispinosa).Methods:The lyophilized C.gigantea flower aqueous extract of 50 mg was added with 100 mL of TiO(OH_2)(10 mM)and magnetically stirred for 6 h.Synthesized TiO_2 NPs were characterized by X-ray diffraction(XRD).Fourier transform infrared spectroscopy(FTIR),Scanning electron microscopy(SEM),and Energy dispersive X-ray spectroscopy(EDX).The synthesised TiO_2 NPs were tested against the larvae of R(B.)microplus and adult of H.bispinosa were exposed to filter paper impregnated method.Results:XRD confirmed the crystalline nature of the nanoparticles with the mean size of 10.52 nm.The functional groups for synthesized TiO_2NPs were 1405.19,and 1053.45 cm^(-1)for-NH_2 bending,primary amines and amides and 1053.84and 1078.45 cm^(-1)for C-O.SEM micrographs of the synthesized TiO_2 NPs showed the aggregated and spherical in shape.The maximum efficacy was observed in the aqueous flower extract of C.gigantea and synthesized TiO_2 NPs against R.(B.)microplus(LC_(50)=24.63 and 5.43 mg/L and r^2=0.960 and 0.988)and against H.bispinosa(LC_(50)=35.22 and 9.15 mg/L and r^2=0.969 and 0.969).respectively.Conclusions:The synthesized TiO_2 NPs were highly stable and had significant acaricidal activity against the larvae of R.(B.)microplus and adult of H.bispinosa.This study provides the first report of synthesized TiO_2 NPs and possessed excellent anti-parasitic activity.

Phytochemical composition and in vitro antioxidant activity of aqueous extract of Aerva lanata(L.) Juss.ex Schult.Stem(Amaranthaceae)

Objective:To analyze the phytochemical composition and in vitro antioxidant properties of aqueous extract of Aerva Lanata（A.lanata） stem.Methods:During the preliminary phytochemical analysis,the aqueous extract of A.Ianata was screened for the presence of carbohydrates,proteins,phenolic compounds,oil and fats,saponins,flavonoids,alkaloids. tannins and phytosterols.Antioxidant activity of the extract was determined by 2.2-dipbenyl- 1-picrylhydrazyl radical scavenging activity,metal chelating activity,reducing power activity and DNA damage inhibition activity.Analysis of phenolic compounds was performed by FolinCiocaiteau reagent method and gradient high performance liquid chromatography technique. Results:Preliminary phytochemical analysis exhibited the presence of phenolic compounds, saponins.flavonoids.tannins and phytosterols as major phytochemical groups.The extract exhibited high 2.2—diphenyl-1-picrylhydrazyl radical scavenging activity(IC50= 110.74μg/ mL).metal chelating activity(IC50= 758.17μg/mL).reducing power activity and DIA damage inhibition efficiency.The extract was reported to possess a high amount of total phenolic content and some of them were identified as gallic acid（3,4,3-OH）,apigenin-7—O-glucoside tapigetrin), quercetin-3—O-rutinoside（rutin） and myricetin（3,5,7,3,4,5-OH）by high performance liquid chromatography analysis.The extract was found non toxic towards human erythrocytes in the hemolytic assay(IC50= 24.89 mg/mL).Conclusions:These results conclud that A.lanata stem possesses high antioxidant activity and can he used for the development of natural and sale antioxidant compounds.

Influence of Substrate Feeding and Process Parameters on Production of Coenzyme Q₁₀Using <i>Paracoccus denitrificans</i>ATCC 19367 Mutant Strain P-87

Coenzyme Q10 (CoQ10), an important antioxidant molecule playing a major role in electron transport chain, has been commercially produced by fermentation process for the use in oral nutraceutical formulations. Constructing the high-yielding CoQ10 producing strains is a pre-requisite for cost-effective production. A superior mutant strain P-87 generated from Paracoccus denitrificans ATCC 19367, which showed 1.25-fold improvement in specific CoQ10 content higher than the wild type strain at shake flask level, was selected to carry out the studies on CoQ10 yield improvement through fermenter process optimization. In the course of study, initially the cane-molasses-based medium and fed-batch fermentation strategy using pHBA in combination with sucrose were standardized in shake flask using wild type strain. This strategy was subsequently translated at 2 L laboratory fermenter while optimizing the fermentation process parameters using improved mutant strain P-87. Under optimized fermentation condition, mutant strain P-87 produced 49.85 mg/L of CoQ10 having specific content of 1.63 mg/g of DCW, which was 1.36 folds higher than the specific CoQ10 content of wild-type strain under similar optimized condition. The temperature and DO were found to be critical parameters for CoQ10 production by mutant strain P-87. The optimum temperature was found to be 32°C and the optimum DO concentration to be maintained throughout the fermentation cycle was found to be 30% of air saturation. Overall, a new cost-effective process has been established for the production of CoQ10 using the cheaper substrate “cane molasses” and higher CoQ10 producing mutant strain P-87.

Antioxidant and hepatoprotective potentials of novel endophytic fungus Achaetomium sp.,from Euphorbia hirta

Objective: To isolate, identify and evaluate the antioxidant, antimicrobial, hepatoprotective potentials, total phenolic content, flavonoid content, tannin content of ethyl acetate extract of endophytic fungus Achaetomium sp., isolated from Euphorbia hirta. Methods: Hepatoprotectivity of ethyl acetate extract of Achaetomium sp., was evaluated by CCl4 induced toxicity in Hep G2 cells and subsequently analyzed for cell viability using MTT assay. It also demonstrates antioxidant and antimicrobial potentials by DPPH radical scavenging assay and well diffusion assay respectively. Quantification of total phenolic content, tannin content and flavonoid content were assessed by spectroscopic methods. Results: Phenols, flavonoids and tannins were the phytochemicals present in ethyl acetate extract of Achaetomium sp., with rich phenolic content exhibited potent hepatoprotective, antimicrobial and antioxidant activities. The hepatoprotective activity was recorded as of 72.13%±2.948% of cell viability at a concentration of 150 μg/m L, whereas the standard silymarin showed 93.260%±0.784%. It was observed to be dose dependent, when CCl4 exposed Hep G2 cells were treated with different concentrations of ethyl acetate extract. Antibacterial activity showed significant inhibition against Staphylococcus aureus, Pseudomonas aeroginosa, and Klebsiella pneumonia. The antioxidant activity ranged from 66.890%±1.385% to 87.340%±0.289% with(44.02±1.57) μg of total phenolics,(54.54±1.82) μg of flavonoid content and(18.790±1.018) μg of tannin content. Ascorbic acid, BHT(butylated hydroxyl toluene) Gallic acid and Pyrogallol were used as standards which showed 98.370%±0.763%; 97.080%±0.636%; 94.890%±1.103% and 96.980%±0.098% reducing potential respectively. Conclusions: The results reveal that the metabolites produced by endophytic fungi isolated from Euphorbia hirta could be novel natural products that could lead to new drug discovery.

In-vitro antimicrobial activity of marine actinobacteria against multidrug resistance Staphylococcus aureus

Objective:To investigate the antibacterial aclivily of marine actinobacteria against multidrug resistance Staphylococcus aureus(MDRSA).Methods:Fifty one actinobacterial strains were isolated from salt pans soil,costal area in Kothapattanam,Ongole,Andhra Pradesh.Primary screening was done using cross-streak method against MDRSA.The bioaclive compounds are extracted from efficient actinobacteria using solvent extraction.The antimicrobial activity of crude and solvent extracts was perfomied using Kirby-Bauer method.MIC for ethyl acetate extract was determined by modified agar well diffusion method.The potent actinobacteria are identified using Nonomura key,Shirling and Gottlieb 1966 with Bergey's manual of determinative bacteriology.Results:Among the fifty one isolates screened for antibacterial activity,SRB25were found efficient against MDRSA.The ethyl acetate extracts showed high inhibition against test organism.MIC test was performed with the ethyl acetate extract against MDRSA and found to be 1 000μg/mL.The isolaled actinobacteria are identified as Streptomyces sp with the help of Nonomura key.Conclusions:The current investigation reveals that the marine actinobacteria from salt pan environment can be able to produce new drug molecules against drug resistant microorganisms.

Antioxidant activity of newly discovered lineage of marine actinobacteria

Objective:To investigate the antioxidant activity of marine actinobacteria.Methods:The content of total phenolics,the level of antioxidant potential by DPPH radical scavenging activity,metal chelating activity,FRAP method,βcarotene assay and NO scavenging activity in extract were determined.Results:In all the methods the extract exhibited good scavenging activity except NO scavenging activity.The IC50 values of marine actinobacteria extract on DPPH radical were found to be 41.09μg/mL.The zone of color retention was 12 mm inβ-carotene bleaching assay. DNA protective efficiency of the extracts was also studied using UV- photolysed H2O2-driven oxidative damage to pBR322.HPLC analysis identified some of the major phenolic compounds in extracts,which might he responsible for the antioxidant potential and cyto-protection.It showed a 100%cytotoxic effect in brine shrimp lethality assay within 10 mins.The novel actinobacteria was identified as Streptomyces LK-3（JF710608） through I6S rDNA Sequencing.Conclusions:The results obtained suggest that the extracts bear anti-cancer metabolites and could be considered as a potential source for anti-cancer drug development.

Hyperbaric oxygen therapy and chemokine administration - a combination with potential therapeutic value for treating diabetic wounds

Non-healing wounds impart serious medical problems to people with diabetes.Amongst 15%of diabetic patients,the incidence of foot ulcer is the most prevailing,which confers a significant risk of limb amputation,mainly due to hypoxia and impairment in cell signaling.Alteration in the expression of chemokines and the related factors in diabetic conditions delays the recruitment of different cell types,including fibroblasts,keratinocytes,and immune cells such as macrophages to the site of injury,further impairing neovasculogenesis,reepithelialization,and extracellular matrix formation.Thus,proper activation of effector cells through an accurate signal pathway is necessary for better therapeutic application.Hyperbaric oxygen therapy(HBOT)is the current treatment prescribed by medical practitioners,shown to have increased the wound healing rate by reducing the need for significant amputation among the diabetic population.However,the risk of morbidity associated with HBOT needs complete attention through rigorous research to avoid adverse outcomes.Altering the level of pro-angiogenic chemokines may regulate the inflammatory response,further promote vascularization,and enhance the complete healing of wounds in diabetic patients.Thus,a combination of better therapeutic approaches could pave the way for developing a successful treatment for diabetic foot and wound healing.

Laxative Effect of Eugenia Jambolana Crude Leaf Bud Extract

The therapeutic value of Eugenia jambolana Lam. commonly known as ‘Jamun’ has been recognized in different system of traditional medication for the treatment of different diseases and ailments. It contains several phytoconstituents belonging to category of alkaloids, glucosides, flavonoides and volatile oils. It has been reported as digestive, astringent to the bowels, anthelmintic, sore throat, bronchitis, asthma, thirst, biliousness, dysentery, blood purifier, ulcers and diabetes. There are few reports available on clinical uses of Eugenia jambolana in diabetes that have shown promising results. In south India ayurvedic practitioners were using the leaf buds of Eugenia jambolana to induce laxative effect and to clean up the intestinal contents before starting any medication. The result showed that of E. jambolana stimulates the contractile action of frog and mice through an acetylcholine-like mechanism and effectively stimulates gastrointestinal motility in mice and frogs. In this paper we have discussed the laxative effect of Eugenia jambolana leaf bud extract which was never reported scientifically.

Molecular basis of fluoride toxicities:Beyond benefits and implications in human disorders

Detrimental impacts of fluoride have become a global concern for several decades.Despite its beneficial role which is restricted only in skeletal tissues,deleterious effects are also observed in soft tissues and systems.The generation of enhanced oxidative stress is the commencement of excess fluoride exposure which may lead to cell death.Fluoride causes cell death through autophagy via Beclin 1 and mTOR signaling pathways.Beside these,several or-gan specific anomalies through different signaling pathways have been documented.Mitochon-drial dysfunction,DNA damage,autophagy and apoptosis are the damaging outcomes in case of hepatic disorders.Urinary concentration defects and cell cycle arrest have been reported in renal tissues.Abnormal immune response has been characterized in the cardiac system.Cogni-tive dysfunction,neurodegenerative condition and learning impairment have also been observed.Altered steroidogenesis,gametogenic abnormalities,epigenetic alterations and birth defect are the major reprotoxic conclusions.Abnormal immune responses,altered immu-nogenic proliferation,differentiation as well as altered ratio of immune cells are well-defined anomalies in the immune system.Though the mechanistic approach of fluoride toxicity in phys-iological systems is common,it follows different signaling cascades.This review emphasizes diverse signaling pathways which are the targets of overexposed fluoride.

The journey of gut microbiome - An introduction and its influence on metabolic disorders

BACKGROUND： Metabolic disorders such as Obesity, Diabetes Type 2 （T2DM） and Inflammatory Bowel Diseases （IBD） are the most prevalent globally. Recently, there has been a surge in the evidence indicating the correlation between the intestinal microbiota and development of these metabolic conditions apart from predisposing genetic and epigenetic factors. Gut microbiome is pivotal in controlling the host metabolism and physiology. But imbalances in the microbiota patterns lead to these disorders via several pathways. Animal and human studies so far have concentrated mostly on metagenomics for the whole microbiome characterization to understand how microbiome supports health in general. However, the accurate mechanisms connecting the metabolic disorders and alterations in gut microbial composition in host and the metabolites employed by the microorganisms in regulating the metabolic disorders is still vague. OBJECTIVE： The review delineates the latest findings about the role of gut microbiome to the pathophysiology of Obesity, IBD and Diabetes Mellitus. Here, we provide a brief introduction to the gut microbiome followed by the current therapeutic interventions in restoration of the disrupted intestinal microbiota. METHODS： A methodical PubMed search was performed using keywords like ＂gut microbiome,＂ ＂obesity, diabetes,＂ ＂IBD,＂ and ＂metabolic syndromes.＂ All significant and latest publications up to January 2018 were accounted for the review. RESULTS： Out of the 93 articles cited, 63 articles focused on the gut microbiota association to these disorders. The rest 18 literature outlines the therapeutic approaches in maintaining the gut homeostasis using probiotics, prebiotics and faecal microbial transplant （FMT）. CONCLUSION： Metabolic disorders have intricate etiology and thus a lucid understanding of the complex host-microbiome inter-relationships will open avenues to novel therapeutics for the diagnosis, prevention and treatment of the metabolic diseases.

Computational Identification of miRNAs and Their Target Genes from Expressed Sequence Tags of Tea(Camellia sinensis)

MicroRNAs （miRNAs） are a newly identified class of small non-protein-coding post-transcriptional regulatory RNA in both plants and animals. The use of computational homology based search for expressed sequence tags （ESTs） with the Ambros empirical formula and other structural feature criteria filter is a suitable combination towards the discovery and isolation of conserved miRNAs from tea and other plant species whose genomes are not yet sequenced. In the present study, we blasted the database of tea （Camellia sinensis） ESTs to search for potential miRNAs, using previously known plant miRNAs. For the first time, four candidate miRNAs from four families were identified in tea. Using the newly identified miRNA sequences, a total of 30 potential target genes were identified for 11 miRNA families; 6 of these predicted target genes encode transcription factors （20%）, 16 target genes appear to play roles in diverse physiological processes （53%） and 8 target genes have hypothetical or unknown functions （27%）. These findings considerably broaden the scope of understanding the functions of miRNA in tea.

对香豆酸对佐剂性关节炎大鼠的抗过氧化作用(英文)

目的:研究对香豆酸,一种食物中含有的酚酸对佐剂性关节炎大鼠的抗过氧化治疗作用。方法:大鼠右后肢脚垫皮内注射0.1mL弗氏完全佐剂诱发关节炎。于佐剂注射后第11～18天连续给药8d,治疗组用药为对香豆酸(100mg/kg大鼠体质量,腹腔内给药),阳性对照组药物为吲哚美辛(3mg/kg大鼠体质量,腹腔内给药)。对佐剂性关节炎大鼠的肝脏、脾脏、血浆内脂质过氧化作用水平和抗氧化水平的变化进行测量,同时分析大鼠后肢的影像学改变以衡量对香豆酸的抗氧化作用。结果:佐剂性关节炎大鼠经对香豆酸治疗后,脂质过氧化作用和抗氧化水平恢复近正常值。影像学观察发现,治疗组大鼠的软组织肿胀、骨侵蚀及关节间隙狭窄情况得到缓解。结论:对香豆酸对佐剂性关节炎大鼠有明显的治疗作用,这种治疗作用可能与其抗氧化作用相关。

生姜提取物6-姜酚对对乙酰氨基酚致小鼠肝脏毒性的保护作用(英文)

目的:研究生姜提取物6-姜酚对对乙酰氨基酚致小鼠肝脏毒性的保护作用。方法:实验组小鼠在腹腔注射对乙酰氨基酚(900 mg/kg)30 min后,分别给予6-姜酚(30 mg/kg)或标准对照药水飞蓟素(25 mg/kg)。注射对乙酰氨基酚4 h后处死小鼠,检测血清中天冬氨酸氨基转移酶、丙氨酸氨基转移酶、碱性磷酸酶活性,总胆红素的含量及肝匀浆中脂质过氧化及抗氧化情况,如超氧化物歧化酶、过氧化氢酶、谷胱甘肽过氧化物酶、谷胱甘肽还原酶、谷胱甘肽转移酶活性及还原型谷胱甘肽含量。结果:与对照组相比,6-姜酚及水飞蓟素均能显著降低对乙酰氨基酚引起的小鼠血清天冬氨酸氨基转移酶、丙氨酸氨基转移酶、碱性磷酸酶活性及总胆红素含量的升高(P<0.05)。此外,6-姜酚及水飞蓟素有效控制了肝脏内丙二醛的形成及各种抗氧化酶的降低(P<0.05)。结论:本研究的结果证实了6-姜酚具有与水飞蓟素相当的保肝作用。

Evaluating the effectiveness of marine actinobacterial extract and its mediated titanium dioxide nanoparticles in the degradation of azo dyes

Aim of the present study was to synthesize titanium dioxide nanoparticles （YiO2 NPs） from marine actinobacteria and to develop an eco-friendly azo-dye degradation method. A total of five actinobacterial isolates were isolated from Chennai marine sediments, Tamilnadu, India and analyzed for the synthesis of TiO2 NPs using titanium hydroxide. Among these, the isolate PSV 3 showed positive results for the synthesis of TiO2 NPs, which was confirmed by UV analysis. Further characterization of the synthesized TiO2 NPs was done using XRD, AFM and FI＇-IR analysis. Actinobacterial crude extract and synthesized TiO2 NPs was found efficient in degrading azo dye such as Acid Red 79 （AR-79） and Acid Red 80 （AR-80）. Degradation percentage was found to be 81% for AR-79, 83% for AR-80 using actinobacterial crude extract and 84% for AR-79, 85% for AR-80 using TiO2 NPs. Immobilized actinobacterial ceils showed 88% for AR-79 and 81% for AR- 80, dye degrading capacity. Degraded components were characterized by FT-IR and GC-MS analysis. The phytotoxicity test with 500 μg/mL of untreated dye showed remarkable phenotypic as well as cellular damage to Tagetes erecta plant. Comparatively no such damage was observed on plants by degraded dye components. In biotoxicity assay, treated dyes showed less toxic effect as compared to the untreated dyes.

Exploring the bioactive potential of Serriatia marcescens VITAPI （Acc： 1933637） isolated from soil

BACKGROUND： Serratia is one of the most important groups of bacteria which produces proteolytic enzymes effectively and known to possess anti-inflammatory properties. The main focus of the current study was to extract the enzyme serratiopeptidase and pigment prodigiosin from Serratia mascescens. Prodigiosin is a red colored pigment produced by the bacterium Serratia marcescens. It is emerging as a valuable molecule because of its large applications. It has already been proved that pigmented strain of Serratia marcescens is less virulent than non-pigmented strains. Moreover the strain we have obtained is from farm soil which indicates that prodigiosin production can be carded safely using this strain. METHODS： In the present study, the isolate VITASP strain was confirmed by morphological, biochemical and molecular studies. The enzyme and pigment were analyzed for anti-oxidant, anti-inflammatory and cytotoxic properties. RESULTS： The isolate was further confirmed and identified as Serratia marcescens with 99% similarity. The extracted pigment showed potent radical scavenging effect with 86% and the enzyme was found to inhibit 83%, which was significant in comparison to ascorbic acid standard. The in vitro anti-inflammatory effect of pigment in controlled experimental conditions revealed its protection at 88% and the enzyme with 90%. Aspirin was used as the reference drug. The present findings exhibited a concentration dependent inhibition. The cytotoxic bioassay of pigment showed the IC5o value as （50） ~tg/mL with 63% cytotoxicity which was statistically significant compared to positive control. CONCLUSIONS： Therefore, it appears to be an essential remedial and application research. It may turn out to be highly beneficial to mankind in solving many problems associated with human health.

Isolation and characterization of tyrosinase produced by marine actinobacteria and its application in the removal of phenol from aqueous environment

The present study was focused on screening and characterization of tyrosinase enzyme produced by marine actinobacteria and its application in phenolic compounds removal from aqueous solution. A total of 20 strains were isolated from marine sediment sample and screened for tyrosinase production by using skimmed milk agar medium. Among 20 isolates, two isolates LK-4 and LK-20 showed zone of hydrolysis and these were taken for secondary screening by using tyrosiue agar medium. Based on the result of secondary screening LK-4 was selected for further analysis, such as tyrosinase assay, protein content and specific activity of the enzyme. The tyrosinase enzyme was produced in a SS medium and was partially purified by ammonium sulfate precipitation, dialysis and SDS PAGE. The isolate （LK-4） was identified as Streptomyces espinosus using 16S rRNA gene sequencing and named as ＂Streptomyces espinosus strain LK4 （KF806735）＂. The tyrosinase enzyme was immobilized in sodium alginate which was applied to remove phenolic compounds from water. The enzyme efficiently removed the phenolic compounds from aqueous solution within few hours which indicated that tyrosinasc enzyme produced by Streptomyces espinosus strain LK-4 can be potently used for the removal of phenol and phenolic compounds from wastewater in industries.