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Inflammation remains a complex process in host defense system and the pathology caused by it remains unsolved till date. Several steroidal and non-steroidal anti-inflammatory drugs in current use to treat inflammatory diseases elicit side effects in longer run. Hence, in order to counteract these outcomes, a pharmacognistic study using a Lamiaceae plant, Plectranthus amboinicus has been focused in the present study. P. amboinicus commonly known as Indian borage has been used in folk medicine for years in treating numerous diseases. It is well known for the presence of major phytochemical constituents which possess anti-inflammatory properties. Assessment of biochemical constituents and molecular mechanisms behind the anti-inflammatory property of P. amboinicus leaves were carried out in in vivo and in vitro conditions. In vivo studies of formalin-induced nociception and paw edema in mice and for in vitro condition lipopolysaccharide-induced inflammation using IC-21 macrophage cells were performed. Aqueous and ethyl acetate leaf extracts of P. amboinicus were analyzed for its phytochemical constituents. Variations in phytochemical components were noticed in GC-MS analysis. The carvacrol containing ethyl acetate extract exhibited higher anti-nociception in terms of lower paw licking time in the later phase of nociception and lowered hind paw edema volume up to 35%. The biochemical analysis revealed that the oxidative stress markers such as malondialdehyde and antioxidant enzymes have been modulated upon pretreatment with both extracts and also modulated the expression of inducible nitric oxide synthase, cyclooxygenase 2, Interleukin-1β, histamine 1 receptor genes and nuclear factor kappa B protein. In addition, nitric oxide inhibitory effect was observed in P. amboinicus treated IC-21 macrophages. Therefore, the molecular mechanism put forth in this study provides insights that the presence of carvacrol in ethyl acetate extract enhances anti-inflammatory activity.

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.

Cryptococcus neoformans, an encapsulated fungal pathogen is evolving as a major threat to immune-compromised patients and rarely to healthy individuals also. The cell wall bound capsular polysaccharide, melanin pigment and biofilm formation are major virulence factors that are known to contribute to cryptococcal meningitis. In the present study, a furanone derivative, (E)-5-benzylidenedihydrofuran-2(3H)-one (compound-6) was evaluated against biofilm of seven different strains of C. neoformans in melanized and non-melanized condition. In addition, the efficacy of compound-6 in activation of TLR-2, opsonophagocytosis, and modulation of cytokine expression during phagocytosis were studied. During the biofilm study, we found that moderate capsule size favored biofilm formation. Interestingly, the minimum biofilm eradication concentration (MBEC0.5) of melanized biofilm was found to be achieved at 1- to 1.7-fold higher MBEC0.5 of non-melanized cells. The maximum eradication of 77% and 69% of non-melanized and melanized biofilm were observed. The capsule size was reduced to half of its size with marked changes in morphology. Furthermore, expression of TLR2, iNOS and pro-inflammatory cytokines such as TNF-α, IL-12, and IFN-γ were also facilitated by compound-6. The correlation analysis showed a positive correlation between phagocytosis and the expression of TLR-2, iNOS, IL-6, IL-12. Collectively, the significant effect of compound-6, anti-melanization activity, antibiofilmand effective immunomodulant could be an interesting dual strategy drug agonist against cryptococcal meningitis.

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.

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.

The emergence and spread of pathogens harboring extended spectrum beta-lactamase (ESBL) like carbapenem resistant Gram negative bacteria are the major emerging threat to public health. Of particular concern Klebsiella pneumoniae carbapenamase- producing strains have been recorded worldwide. Catheter associated urinary tract infections (CAUTI) caused by K. pneumoniae are significantly associated with morbidity and mortality. Hence the present work was aimed to develop a strategy for addressing these issues through an innovative approach of antibiofilm and immunomodulation. These two independent activities were analyzed in a Streptomyces derived ASK2 bioactive compound. While analysing the effect of sub-minimum inhibitory concentrations (sub-MICs), 0.5x of Minimum Inhibitory Concentration (MIC) was found to be more effective in preventing biofilm formation on coverslip and silicone catheter. The minimum biofilm eradication concentration (MBEC) was found to be 15-fold higher MIC with eradication of 75% of 3 day old biofilm. Apart from its antibiofilm potential, ASK2 also acts as an opsonin and enhances phagocytic response of macrophages against multidrug resistant K. pneumoniae. In addition, ASK2 resulted in elevated levels of nitric oxide generation by the macrophages and has a stimulating effect on IL-12, IFN-γ, and TNF-α proinflammatory cytokines. The opsonic role of ASK2 and its potential in modulating proinflammatory cytokines secreted by macrophages implies the importance of ASK2 in modulating cellular immune response of macrophages against MDR K. pneumoniae. The present study proposes ASK2 as a promising candidate for treating MDR K. pneumoniae infections with its dual properties of antibiofilm and immunomodulatory activities.

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.

Made available in DSpace on 2018-11-26T17:06:11Z (GMT). No. of bitstreams: 0 Previous issue date: 2016-11-01 University Grants Commission Department of Biotechnology, New Delhi, India UGC, New Delhi, India Hemocyanins are large extracellular respiratory proteins distributed within the hemolymph of arthropods, molluscs and larval stages of certain insects. In the present study, we characterized the humoral immune functions of cation specific antibacterial hemocyanin (Ab-Hcy) from mud crab, S. serrata. The bacteriolytic activity of Ab-Hcy was against pathogenic and non-pathogenic to crustaceans which includes, Bacillus sp. N1, B. flexus N3, E. coli, P. aeruginosa, V. harveyi, V. parahaemolyticus and V. vulnificus and also expressed bacteriostatic activity against E. coli, B. flexus N3 and V. harveyi. The agglutination activity of Ab-Hcy ranged from 4-16 against B. flexus N3, E. coli, V. harveyi and V. vulnificus and Ab-Hcy also agglutinated with rat and human O erythrocytes (HA titer: 2). The Ab-Hcy lysed the panel of mammalian erythrocytes and strong hemolytic activity of 1.40, 1.28 and 1.22 units.min(-1) . mg protein(-1) against human A, B and O erythrocytes respectively. It was also found that the Ab-Hcy possess phenoloxidase (PO) activity by oxidizing L-DOPA. These results suggest that Ab-Hcy from the serumofmud crab, S. serrata, is capable of performing multiple humoral immune functions in addition to oxygen transportation. Statement of relevance: This manuscript deals with the humoral immune functions, particularly antibacterial property of hemocyanin in crustacean. In this paper we report the different type of antibacterial activity and other humoral immune functions of a hemocyanin isolated from the serum of mud crab, Scylla serrata. This work will help to understand the role of respiratory molecule in immune system, also will be helpful improve disease free crustacean aquaculture industry. And we also state this research work is relevant and suitable to publish in the Journal Aquaculture. (C) 2016 Elsevier B.V. All rights reserved. Univ Madras, Dept Zool, Guindy Campus, Madras 600025, Tamil Nadu, India Sao Paulo State Univ, Sch Dent Araraquara, Dept Diag & Surg, Rua Humaita 1680, BR-14801903 Sao Paulo, Brazil Sao Paulo State Univ, Sch Dent Araraquara, Dept Diag & Surg, Rua Humaita 1680, BR-14801903 Sao Paulo, Brazil University Grants Commission: 4-3/2006 (BSR)/5-133/2007 Department of Biotechnology, New Delhi, India: BT/PR11998/AAQ/03/462/2009

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.

The genomic DNA from ten isolates of the cattle tick, Boophilus microplus collected in and around Chennai, India, was analyzed by random amplified polymorphic DNA (RAPD) using PCR. Selected five random primers were used for the study of genetic variability among different isolates of B. microplus. A high degree of genetic polymorphism with a different pattern of RAPD profiles for each tick isolate was detected with all these random primers. This variability was also confirmed by similarity coefficient values and dendrogram which were performed using mean RAPD profiles for all the primers between various isolates of ticks. The findings suggest the existence of a complex genotypic diversity of the tick B. microplus in an endemic region such as Chennai.