ANTICARCINOGENIC ACTIVITY OF RICE BRAN PHYTIC ACID AGAINST HUMAN BREAST CANCER CELL LINE (MCF-7)

Phytic acid (PA) has been reported for anti-inflammatory, antioxidant and anticancer activity. However, molecular mechanism of anticancer activity is not clear. This study investigated the anticancer activity of rice bran PA against breast cancer (MCF-7). Cytotoxicty of PA (0 to 7 mM) against MCF-7 cells was examined by MTT and LDH assays after 24 and 48 h treatment. Apoptotic activity was evaluated by expressional analysis of apoptosis-regulatory genes [i.e., p53, Bcl-2, Bax, caspase-3 and -9] by reverse transcriptase-PCR and DNA fragmentation assay. PA inhibited the growth of MCF-7 cells in a concentration dependent manner (p≤0.04). After 48 h treatment, cells viability was recorded 80.9, 71.1, 59.8, 36.6, 26.7 and 15.9% in MTT assay and 85.3, 72.6%, 62.3%, 42.1, 31.7 and 21.7% in LDH assay at concentration of 1.4, 2.2, 3.0, 3.8, 4.6, and 5.4 mM respectively. Hence, treatment of PA for 24 h, recorded viability of cells 84.6, 73.8, 61.0, 47.0, 28.8 and 17.3% in MTT assay and 87.8, 77.5%, 62.9%, 49.8, 35.7 and 23.3% in LDH assay at concentration of 2, 3, 4, 5, 6, and 7 mM, respectively. PA treated MCF-7 cells showed up-regulation of p53, Bax, caspase-3 and -9, and down-regulation of Bcl-2 gene (p ≤0.03). At IC50 (3.4 mM) of PA, the p53, Bax, caspase 3 and -9 genes were up-regulated by 6.34, 4.90, 23.45 and 15.03 folds respectively. Also, the fragmented genomic DNA in PA treated cells showed the signs of apoptosis. Our study endorsed the biological activity of PA and demonstrated the PA induced growth inhibition and apoptosis in MCF-7 cells by modulating the expression of apoptosis-regulatory genes. Keyword: Phytic acid, antioxidant, cytotoxicity, apoptosis, caspases, p53, Bax, Bcl-2, DNA fragmentatio

Anti-Inflammatory Activity of Calotropis gigantea and Tridax procumbens on Carrageenin-Induced Paw Edema in Rats

The anti-inflammatory activities of extract of Calotropis gigantea R.Br. and Tridax procumbens Linn., were assessed on carrageenin-induced paw edema along with standard drug, Ibuprofen. The Ibuprofen significantly reduced paw edema at the dose of 200mg/Kg bw orally. The oral administration equi-effective dose (ED50) of C. gigantea (600mg/Kg bw) and T. procumbens (400 mg/Kg bw) individually revealed about 20-35% more activity than the one rendered by administration of 50mg/Kg bw of Ibuprofen. The effect of C. gigantea and T. procumbens along with various dose regimen of Ibuprofen showed greater anti-inflammatory activities than the Ibuprofen alone

Synthesis and mechanistic studies of diketo acids and their bioisosteres as potential antibacterial agents

A series of diketo esters and their pertinent bioisosteres were designed and synthesized as potent antibacterial agents by targeting methionine amino peptidases (MetAPs). In the biochemical assay against purified MetAPs from Streptococcus pneumoniae (SpMetAP1a), Mycobacterium tuberculosis (MtMetAP1c), Enterococcus faecalis (EfMetAP1a) and human (HsMetAP1b), compounds 3a, 4a and 5a showed more than 85% inhibition of all the tested MetAPs at 100 μM concentration. Compounds 4a and 5a also exhibited antibacterial potential with MIC values 62.5 μg/mL (S. pneumoniae), 31.25 μg/mL (E. faecalis), 62.5 μg/mL (Escherichia coli) and 62.5 μg/mL (S. pneumoniae), 62.5 μg/mL (E. coli), respectively. Moreover, 5a also significantly inhibited the growth of multidrug resistant E. coli strains at 512 μg/mL conc., while showing no cytotoxic effect towards healthy CHO cells and thus being selected. Growth kinetics study showed significant inhibition of bacterial growth when treated with different conc. of 5a. TEM analysis also displayed vital damage to bacterial cells by 5a at MIC conc. Moreover, significant inhibition of biofilm formation was observed in bacterial cells treated with MIC conc. of 5a as visualized by SEM micrographs. Interestingly, 5a did not cause an alteration in the hemocyte density in Galleria mellonella larvae which is considered in vivo model for antimicrobial studies and was non-toxic up to a conc. of 2.5 mg/mL

Plasmodium vivax lineages: geographical distribution, tandem repeat polymorphism, and phylogenetic relationship

<p>Abstract</p> <p>Background</p> <p>Multi-drug resistance and severe/complicated cases are the emerging phenotypes of vivax malaria, which may deteriorate current anti-malarial control measures. The emergence of these phenotypes could be associated with either of the two <it>Plasmodium vivax </it>lineages. The two lineages had been categorized as Old World and New World, based on geographical sub-division and genetic and phenotypical markers. This study revisited the lineage hypothesis of <it>P. vivax </it>by typing the distribution of lineages among global isolates and evaluated their genetic relatedness using a panel of new mini-satellite markers.</p> <p>Methods</p> <p><it>18S SSU rRNA S-type </it>gene was amplified from 420 <it>Plasmodium vivax </it>field isolates collected from different geographical regions of India, Thailand and Colombia as well as four strains each of <it>P. vivax </it>originating from Nicaragua, Panama, Thailand (Pak Chang), and Vietnam (ONG). A mini-satellite marker panel was then developed to understand the population genetic parameters and tested on a sample subset of both lineages.</p> <p>Results</p> <p><it>18S SSU rRNA S-type </it>gene typing revealed the distribution of both lineages (Old World and New World) in all geographical regions. However, distribution of <it>Plasmodium vivax </it>lineages was highly variable in every geographical region. The lack of geographical sub-division between lineages suggests that both lineages are globally distributed. Ten mini-satellites were scanned from the <it>P. vivax </it>genome sequence; these tandem repeats were located in eight of the chromosomes. Mini-satellites revealed substantial allelic diversity (7-21, <it>AE </it>= 14.6 ± 2.0) and heterozygosity (<it>He </it>= 0.697-0.924, <it>AE </it>= 0.857 ± 0.033) per locus. Mini-satellite comparison between the two lineages revealed high but similar pattern of genetic diversity, allele frequency, and high degree of allele sharing. A Neighbour-Joining phylogenetic tree derived from genetic distance data obtained from ten mini-satellites also placed both lineages together in every cluster.</p> <p>Conclusions</p> <p>The global lineage distribution, lack of genetic distance, similar pattern of genetic diversity, and allele sharing strongly suggested that both lineages are a single species and thus new emerging phenotypes associated with vivax malaria could not be clearly classified as belonging to a particular lineage on basis of their geographical origin.</p

Enhanced anti-tumor efficacy of paclitaxel with PEGylated lipidic nanocapsules in presence of curcumin and poloxamer: In vitro and in vivo studies

International audienceCancer chemotherapeutic drug containing PEGylated lipidic nanocapsules (D-LNCs) were formulated by the controlled addition of organic phase (combined solution of paclitaxel and curcumin in a mixture of oleic acid and MPEG2000-DSPE (90:2.5 molar ratio) in acetone) to the aqueous phase (consist of Poloxamer 407 as emulsifying agents and glycerol as a co-solvent) at a temperature of 55-60°C followed by evaporation of organic solvent. The obtained pre-colloidal dispersion of D-LNCs was processed through high pressure homogenization to get more uniformly and nano-sized particles. Effect of concentration of emulsifying agent and process variables of high pressure homogenization (pressure and number of cycles) on average particle size and entrapment efficiency was further investigated by constructing Box-Behnken experimental design to achieve the optimum manufacturing process. D-LNCs were characterized by dynamic light scattering, scanning and transmission electron microscopy, Fourier transform infrared spectroscopy, and differential scanning calorimetry. In vitro release studies showed a sustained release pattern of drug from the PEGylated D-LNCs, whereas in vivo pharmacokinetic studies after a single-dose intravenous (i.v.) administration of paclitaxel (15mg/kg) in Ehrlich ascites tumor (EAT)-bearing female Swiss albino mice showed a prolonged circulation time and slower elimination of paclitaxel from D-LNCs as compared with marketed formulation (Paclitec®). From the plasma concentration vs. time profile, i.v. bioavailability (AUC0-∞) of paclitaxel from D-LNCs was found to be increased approximately 2.91-fold (P<0.001) as compared to Paclitec®. In vitro cell viability assay against MCF-7 and MCF-7/ADR cell lines, in vivo biodistribution studies and tumor inhibition study in EAT-bearing mice, all together prove its significantly improved potency towards cancer therapy

High mobility group box (HMGB) proteins of Plasmodium falciparum: DNA binding proteins with pro-inflammatory activity

High mobility group box chromosomal protein 1 (HMGB1), known as an abundant, non-histone architectural chromosomal protein, is highly conserved across different species. Homologues of HMGB1 were identified and cloned from malaria parasite, Plasmodium falciparum. Sequence analyses showed that the P. falciparum HMGB1 (PfHMGB1) exhibits 45, 23 and 18%, while PfHMGB2 shares 42, 21 and 17% homology with Saccharomyces cerevisiae, human and mouse HMG box proteins respectively. Parasite PfHMGB1and PfHMGB2 proteins contain one HMG Box domain similar to B-Box of mammalian HMGB1. Electrophoretic Mobility Shift Assay (EMSA) showed that recombinant PfHMGB1 and PfHMGB2 bind to DNA. Immunofluorescence Assay using specific antibodies revealed that these proteins are expressed abundantly in the ring stage nuclei. Significant levels of PfHMGB1 and PfHMGB2 were also present in the parasite cytosol at trophozoite and schizont stages. Both, PfHMGB1 and PfHMGB2 were found to be potent inducers of pro-inflammatory cytokines such as TNFα from mouse peritoneal macrophages as analyzed by both reverse transcription PCR and by ELISA. These results suggest that secreted PfHMGB1 and PfHMGB2 may be responsible for eliciting/ triggering host inflammatory immune responses associated with malaria infection

Comparative Study of ZnO-and-TiO₂-Nanoparticles-Functionalized Polyvinyl Alcohol/Chitosan Bionanocomposites for Multifunctional Biomedical Applications

This study aimed to synthesize chitosan/polyvinyl alcohol (CS/PVA)-based zinc oxide (ZnO) and titanium dioxide (TiO2) hybrid bionanocomposites (BNCs) and observe their comparative accomplishment against the skin cancer cell line, A431, and antioxidant potential. CS was blended with PVA to form polymeric films reinforced with the immobilization of ZnO and TiO2 nanoparticles (NPs), separately. The optimization of the BNCs was done via physicochemical studies, viz. moisture content, swelling ratio, and contact angle measurements. The free radical scavenging activity was observed for 1,1-diphenyl-2-picryl-hydrazyl, and the antibacterial assay against the Escherichia coli strain showed a higher zone of inhibition. Furthermore, the anticancer activity of the synthesized BNCs was revealed against the skin cancer cell line A431 under varying concentrations of 50, 100, 150, 200, and 300 μg/mL. The anticancer study revealed a high percent of cancerous cell inhibition (70%) in ZnO BNCs as compared to (61%) TiO2 BNCs in a dose-dependent manner

Nitration of H2B histone elicits an immune response in experimental animals

Histone H2B is an autoantigen that appears in circulation due to altered apoptosis/or insufficient clearance and is likely to be involved in the induction and progression of autoimmune diseases since modified-H2B is immunogenic. Our studies demonstrate that tyrosines of H2B histone spontaneously converts to free and nitrotyrosine bound protein in vivo. Commercially available H2B histone was modified with peroxynitrite in vitro. Modified H2B was found to be more immunogenic than native form in experimental animals. Furthermore, the sera of rabbits were analyzed for the native and modified forms of the H2B histone. The binding specificity of autoantibodies was characterized by competitive enzyme-linked immunosorbent assay (ELISA) and band shift assay. The free 3-nitrotyrosine in systemic lupus erythematosus sera was quantified by high-performance liquid chromatography. Peroxynitrite-modified H2B induced high titre antibodies as compared to native form which were directly proportional to the nitrotyrosine content. Furthermore, the induced antibodies showed specificity towards the immunogen and cross-reacted with tyrosine-nitrated proteins. ELISA showed preferential binding of induced anti-peroxynitrite modified H2B antibodies to modified H2B as compared to native H2B. The present study shows that peroxynitrite modification of self-antigen(s) generates neoepitopes capable of inducing modified-H2B autoantibodies in experimental animals