Arumugam Munusamy

Protective effect of ferulic acid and resveratrol against alloxan-induced diabetes in mice.

The present study was to investigate the effect of ferulic acid and resveratrol on alloxan-induced diabetic mice, through analysis of basic biochemical parameters, enzymic as well as non-enzymic activities, lipid peroxidation and immunohistochemical studies. Alloxan was administered as a single dose (75 mg/kg body weight) to induce diabetes in mice. A dose of ferulic acid (10 mg/kg body weight) and resveratrol (20 mg/kg body weight) were administrated orally, to the alloxan-induced diabetic mice. The levels of basic biochemical markers and lipid peroxidation were significantly (P<0.05) increased in alloxan-induced diabetic mice. The levels of antioxidants were significantly (P<0.05) decreased in liver, kidney and serum. Immunohistochemical studies in alloxan induced mice demonstrated a marked increase in the immunoreactivity of nuclear transcription factor (NF-κB). Treating the diabetic mice with doses of ferulic acid and resveratrol restored the changes in the above parameters analyzed. The present study, showed that ferulic acid and resveratrol exerted antioxidant as well as anti-diabetic effects, consequently alleviate liver, kidney and pancreas damage caused by alloxan-induced diabetes, probably through inhibition of the proinflammatory factor, NF-κB.

Biosynthesis of silver nanoparticles using ethanolic petals extract of Rosa indica and characterization of its antibacterial, anticancer and anti-inflammatory activities.

The present study was aimed at biosynthesis of silver nanoparticles (AgNPs) using ethanolic extract of rose (Rosa indica) petals and testing their potential antibacterial activity using selective human pathogenic microbes, anticancer activity using human colon adenocarcinoma cancer cell line HCT 15 as well as anti-inflammatory activity using rat peritoneal macrophages in vitro. The biologically synthesized AgNPs were also characterized by UV-visible spectroscopy, scanning electron microscopy (SEM), transmission electron microscopy (TEM), Fourier transform infrared spectroscopy (FTIR), energy-dispersive X-ray spectroscopy (EDX) and X-ray diffraction (XRD). The characterized AgNPs showed an effective antibacterial activity against Gram negative (Escherichia coli, Klebsiella pneumoniae) than Gram positive (Streptococcus mutans, Enterococcus faecalis) bacteria. MTT assay, analysis of nuclear morphology, mRNA expression of Bcl-2, Bax and protein expression of caspase 3 as well as 9, indicated potential anticancer activity. In addition, green synthesized AgNPs also attenuated cytotoxicity, nuclear morphology and free radical generation (O2(-) and NO) by rat peritoneal macrophages in vitro. The results of our study show the potential green synthesis of silver nanoparticles in mitigating their toxicity while retaining their antibacterial activities.

Synthesis of silver nanoparticles using Solanum trilobatum fruits extract and its antibacterial, cytotoxic activity against human breast cancer cell line MCF 7

In the present study, we have synthesized silver nanoparticles by a simple and eco-friendly method using unripe fruits of Solanum trilobatum. The aqueous silver ions when exposed to unripe fruits extract were reduced and stabilized over long time resulting in biosynthesis of surface functionalized silver nanoparticles. The bio-reduced silver nanoparticles were characterized by UV-visible spectroscopy, Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), transmission electron microscopy (TEM), energy-dispersive spectroscopy (EDX) and X-ray diffraction (XRD). These biologically synthesized silver nanoparticles were tested for its antibacterial activity against few human pathogenic bacteria including Gram-positive (Streptococcus mutans, Enterococcus faecalis) and Gram-negative (Escherichia coli, Klebsiella pneumoniae) bacteria. In addition, we also demonstrated anticancer activity of these nanoparticles in vitro against human breast cancer cell line (MCF 7) using MTT, nuclear morphology assay, Western blot and RT-PCR expression. These results taken together show the potential applications of biosynthesized silver nanoparticles using S. trilobatum fruits.

Opsonic function of sialic acid specific lectin in freshwater crab Paratelphusa jacquemontii

The sialic acid specific humoral lectin, Pjlec of the freshwater crab Paratelphusa jacquemontii was investigated for its opsonin function with rabbit erythrocyte as target cell for phagocytosis by the crab’s hemocyte. The untreated or trypsin treated erythrocyte induced lectin response after challenge however failed when treated with neuraminidase evidently indicating glycan dependency for elicited immune response. Our observation of in vitro phagocytosis of the erythrocyte untreated or coated with serum, clarified serum appeared to be recognized and engulfed by hemocytes but when coated with isolated lectin Pjlec, the response was elicited. Moreover, with trypsin treated erythrocyte the response remained unchanged but neuraminidase or O-glycosidase treatment eliminated the response reaction. This suggested the sialic acid specific reaction of lectin with the erythrocyte and was essential for recognition to allow the lectin Pjlec to act as an opsonin. The flowcytometry observation affirmed the enhancement of phagocytosis by Pjlec coated hemocyte. The efficiency of in vitro hemolysis of Pjlec coated erythrocyte with hemocyte when compared to untreated erythrocyte with or without hemocyte also established the opsonic function of the lectin. The mechanism of phagocytosis and induction were dependent on specific recognition of the erythrocyte by the multivalent binding site of the lectin protein, and the elicitation of the immune response was a function of the sialoglycan surface. The pathway of the challenge suggested that after entry of nonself recognition by lectin was followed by induction and activation of phagocytosis by opsonic binding of the lectin.

Phenoloxidase activity in humoral plasma, hemocyanin and hemocyanin separated proteins of the giant freshwater prawn Macrobrachium rosenbergii

Hemocyanin is a copper containing protein and its role in the immune function of phenoloxidase (PO) activity was investigated in the giant freshwater prawn Macrobrachium rosenbergii. Hemocyanin, sedimented by ultracentrifugation from the plasma appeared on polyacrylamide gel electrophoresis (PAGE 7%) on Coomassie Brilliant Blue and bathocuproine sulfonic acid stain as four copper containing proteins of molecular masses 50, 60, 114 and 325kDa. Accordingly, on diethylaminoethyl-cellulose anion exchange column hemocyanin separated into four proteins designated as MrHc1, MrHc2, MrHc3 and MrHc4 with electrophoretically (PAGE) determined molecular masses of 60, 114, 50 and 325kDa respectively. The reduction of proteins in sodium dodecyl sulphate (SDS)-PAGE revealed that MrHc1 and 3 were monomeric for 60and 50kDa respectively, MrHc2 dimeric of 56 and 58kDa subunits and MrHc4 appeared with three subunits of 74, 76 and 78kDa. The PO activity was determined in plasma, hemocyanin and the four separated hemocyanin proteins in vitro using L-3,4-dihydroxyphenylalanine (L-DOPA) at pH7.5, 25°C and appeared elicited by exogenous activators such as trypsin, SDS, cell wall components of bacteria and polysaccharide laminarin. This study clearly demonstrated hemocyanin as the major copper containing protein in the plasma of M. rosenbergii with potent PO activity.

Cation metals specific hemocyanin exhibits differential antibacterial property in mud crab, Scylla serrata

Abstract Serum of mud crab, Scylla serrata had antibacterial activity against certain host specific bacteria as well as other known crustacean pathogenic bacteria. The respective antibacterial molecule was isolated by ion-exchange chromatographic method using diethylaminoethyl-cellulose matrix and eluted with gradient increase of sodium chloride (0-0.5 M). There were three major peaks on the elution profile and fractions from second major peak exhibit differential antibacterial response against Bacillus flexus N3, Escherichia coli, Vibrio harveyi and Vibrio vulnificus. The electrophoretic analyses of this fraction resulted in single protein band and the native molecular weight of this protein was found to be approximately 305 kDa. The bathocuproine sulfonic acid staining (a specific stain for copper), copper/protein ratio (0.129%) reveals this isolated antibacterial protein to be hemocyanin. This antibacterial hemocyanin is also capable of binding to cations, namely Ca2+, K+, Mg2+, Mn2+ and Zn2+ apart from carrying Fe2+ ions. This native antibacterial hemocyanin was found to contain three or four subunits with possible molecular size of 70-98 kDa under reducing conditions. These findings indicate that the respiratory protein hemocyanin in the presence of cations functions as humoral immune molecule and the detailed investigation would reveal additional immune functional characteristics of this respiratory protein as well as subunits-specific functions of this antibacterial hemocyanin.

Activation of phenoloxidase activity by humoral lectin in hemocytes of freshwater crab Paratelphusa jacquemontii

The lectin, Pjlec isolated from the hemolymph of the freshwater crab Paratelphusa jacquemontii hemagglutinated (HA) with mice, rabbit and rat erythrocytes. However, the lectin failed to agglutinate neraminidase treated asialylated erythrocytes showing its sialic acid specificity. The poyacyrlamide gel electrophoresis of lectin yielded 310kDa proteins, on sodium sulphate dodecyl (SDS) gel appeared as a tetramer with subunits of 76kDa. The observation of in vitro phagocytosis in granular hemocytes of lectin opsonized rabbit erythrocyte by Transmission electron microscopy (TEM) showed the release of lytic vesicles by exocytosis prior to engulfment. The Pjlec lectin also showed an ability to oxidize L-3, 4 dihydroxyphenylalanine (L-DOPA) and in hemocyte lysate preparation (HLS) was enhanced on reduction with SDS and on proteolytic cleavage with trypsin. The lectin appeared to have a regulatory role in activation of enzyme activity associated with phagocytosis and melanin formation.

Immune response of anti-lectin Pjlec antibody in freshwater crab Paratelphusa jacquemontii

Sialic acid specific lectin Pjlec isolated from serum of the freshwater crab Paratelphusa jacquemontii served as an antigen for the production of immunoglobulin (Ig) in Balb/c mice sera. Enzyme-linked immunosorbent assay (ELISA) of mice anti-sera with Pjlec lectin affirmed the induction and production of antibody. Anti-Pjlec antibody was isolated from the antisera of mice by Protein A Sepharose affinity chromatography and checked for purity by immunoblot with lectin. Mass spectrometry (MS/MS) of papain digethe peptide sequence of antigen binding fragment (Fab) and fragment crystallizable (Fc). Coatingsted anti-Pjlec revealed of anti-Pjlec to the target cell, rabbit erythrocyte failed to enhance in vitro phagocytosis in the crab. However, inoculation of anti-Pjlec in the hemolymph of the crab elicited in vitro phagocytosis. Proteins in hemocyte lysate supernatant (HLS) were separated by electrophoresis failed to immunoblot with Pjlec or anti-Pjlec. Peptide sequences of trypsin digested lectin protein appeared homologous to deuterostome chordate. The protostome crab that lack the ability to synthesize sialic acid however bind to sialic acid a deuterostome sugar to suggest the complexity in innate immune system of invertebrates. The application of lectin and its antibody require further study on application of pathological conditions associated with alterations in sialylated cell surface.

Isolation and analysis of mannose/trehalose/maltose specific lectin from jack bean with antibruchid activity

A lectin with insecticidal property against the stored product pest, Callosobruchus maculatus was successfully isolated from the seeds of Canavalia virosa using standard affinity chromatography. The isolated molecule typically behaved like a lectin in its characteristics. It agglutinated indicator red blood cells (RBC) in its native as well as enzyme treated conditions. The enzyme treated RBC types exhibited a very high hemagglutination (HA) titre values and this property of isolated molecule behaved like arcelin, the lectin-like molecules reported from several species of Phaseolus. As a characteristic feature of a lectin, the isolated molecule effectively inhibited the agglutination of indicator RBC types with simple and complex carbohydrates including glycoproteins. This nature of the isolated molecule also relate with characteristic feature of arcelin isoforms in inhibiting HA activity with complex glycoproteins as reported in many studies. Most interestingly, the present study disclosed trehalose as a potent inhibitor of C. virosa lectin. Therefore, feeding insect pests on the lectin like arcelin could serve as antibiosis factor/anti-insect activity. The molecular characteristics of this isolated molecule and its mass studies too revealed its homology with arcelin, arcelin-1, 2 and 6 isoforms of P. vulgaris and lectin from Canavalia cathartica, C. lineata and C. brasiliensis.

Bio-efficacy of crude leaf extracts of Acalypha fruticosa Forssk. against some agriculturally important insect pests

Abstract Objective To investigate the antifeedant and larvicidal activities of Acalypha fruticosa Forssk. (Euphorbiaceae) ( A. fruticosa ) leaf extracts. Methods Efficacy of various organic solvent extracts of dichloromethane, acetone, dimethyl sulfoxide and aqueous extracts of the leaves of A. fruticosa were evaluated for their feeding inhibition and larvicidal activities against third instar larvae of Leucinodes orbonalis ( L. orbonalis ) , Helicoverpa armigera ( H. armigera ) , Spodoptera litura ( S. litura ) and Earias vittella ( E. vittella ). Antifeedant and larvicidal activities were performed by leaf and fruit discs no-choice method at 0.625%, 1.25%, 2.5% and 5% concentrations. Results The results revealed that 5% concentration of dichloromethane extract had significant antifeedant activity on L. orbonalis (77.1%), H. armigera (66.2%), S. litura (74.8%) and E. vittella (67.2%) followed by acetone, dimethyl sulfoxide and aqueous extracts. The result on larvicidal activity also showed that the 5% concentration of dichloromethane extract exhibited significantly higher larval mortality for L. orbonalis (62.12%), H. armigera (62.14%), S. litura (55.11%) and E. vittella (77.15%) when compared to acetone, dimethyl sulfoxide and aqueous extracts. Conclusions From this study, it is concluded that the dichloromethane extracts of leaves of A. fruticosa could serve as a potential natural pesticide for further exploration of active compounds.