Md. Zafaryab

Comparative Analysis of the Antioxidant Activity of Cassia fistula Extracts

Antioxidant potential of various extracts of Cassia fistula was determined by the DPPH, FRAP, Fe3+ reducing power, and hydrogen peroxide scavenging assay. Methanolic extracts of Cassia fistula showed the highest amount of phenolic and flavonoid content and reducing capacity, whereas hexane extracts exhibited the lowest level of reducing capacity. The order of antioxidant activity in Cassia fistula extracts displayed from higher to lower level as methanolic extracts of pulp, methanolic extracts of seed, hexane extracts of pulp, and hexane extracts of seed. The antioxidant potential of Cassia fistula extracts significantly correlated (P < 0.02) with the phenolic content of the methanolic extracts. Ascorbic acid taken as control showed highest antioxidant power in the present study.

Biocompatible nanostructured magnesium oxide-chitosan platform for genosensing application

A novel organic-inorganic platform comprising of chitosan (CH) modified nanostructured magnesium oxide (nanoMgO) has been electrophoretically deposited on the indium-tin-oxide (ITO) substrate. The single stranded probe DNA (ssDNA) sequence of Vibrio cholerae has been covalently functionalized onto CH-nanoMgO/ITO surface. The cytotoxicity assay of nanoMgO particles, examined using human intestinal cell line (INT 407), reveals no significant cytotoxicity at the given doses in the range of 50-350 μg/mL. The X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FT-IR) and various microscopic techniques have been employed for the structural and morphological analysis of the fabricated electrodes. The electrochemical response studies of ssDNA and fragmented genomic DNA hybridized electrode (dsGDNA/CH-nanoMgO/ITO) have been carried out using cyclic voltammetry (CV) and differential pulse voltammetry (DPV) techniques. The dsGDNA/CH-nanoMgO/ITO bioelectrode exhibits a linear response in the range 100-500 ng/μL with improved sensitivity of 36.72 nA/ng/cm(2), faster response time of 3s and high stability of 3-4 months under refrigerated condition. The lower detection limit of fabricated genosensor has been estimated as 35.20 ng/μL and it shows good reproducibility/repeatability.

Phytochemical Composition of Cassia fistula Fruit Extracts and its Anticancer Activity Against Human Cancer Cell Lines

Abstract Cassia fistula Linn. (Fabaceae) fruit has been used in folk medicine and reported for various pharmacological properties. We evaluated the anticancer activity of C. fistula fruit extracts [ethyl acetate extract of pulp (EEP), n-butanol extract of pulp (BEP), ethyl acetate extract of seeds (EES), n-butanol extract of seeds (BES)] against human cervical cancer (SiHa) and breast cancer (MCF-7) cell lines. Phytochemicals analysis revealed that the EEP and EES contained anticancer compounds like 2(3H)-furanone, rhein, thymoland oleic acid. BES contained inositol and palmitic acid, whereas BEP contained inositol and 2-pyrrolidone. IC50 of EEP, EES, BEP and BES against SiHa cells were 415.5±0.19, 435.7±0.11, 535.3±0.32 and 580.2±0.41 µg/mL, whereas against MCF-7 cells were 422.2±0.32, 451.4±0.27, 564.5±0.21 and 575.6±0.47 µg/mL respectively (p ≤ 0.01). At IC50, EEP, EES, BEP and BES treated MCF-7 cells formed colonies of 17.5, 16.3, 22.5 and 18.8 % whereas treated SiHa cells formed colonies of 11.5, 12.8, 19.2 and 16.7 % (p≤0.03). Seeds and pulp treated both cell lines showed up-regulation of p53 and Bax genes, down-regulation of Bcl-2 gene and increased caspase-3, 7 & 10 and -9 enzymes activities. Also, the fragmented genomic DNA in pulp and seeds extracts treated SiHa and MCF-7 cells showed sign of apoptosis.We concluded that the fruit pulp and seeds extracts inhibited MCF-7 and SiHa cells growth and induced cells death by modulating the expression of apoptosis-regulatory genes and caspase enzymes.

Characterization and anti-proliferative activity of curcumin loaded chitosan nanoparticles in cervical cancer.

In the present study the chitosan nanoparticles (CsNPs) and curcumin loaded chitosan nanoparticles (CLCsNPs) were synthesized by tripolyphosphate (TPP) cross-linking method. The nanoparticles were prepared within a zone of appropriate chitosan and TPP concentrations. The average size of CsNPs and CLCsNPs were approximately 189±11.8nm and 197±16.8nm, exhibited a zeta potential of +76±5.6mV and +71±6.4mV respectively and drug entrapment efficiency was ≈85%. The CLCsNPs and CsNPs were further characterized by different physicochemical methods like transmission electron microscopy (TEM), dynamic light scattering (DLS), HPLC, MALDI-TOF, FT-IR, XRD and UV-vis Spectroscopy. In vitro studies revealed a fast release of ≈35% at pH 5 and ≈25% at pH 7.4 of the drug during the first 3h, followed by controlled release of curcumin over a period of 120h and sustained anti-proliferative activity of the drug in a dose and time dependent manner of CLCsNPs and combination with methyl jasmonate. The higher cytotoxicity effect of CLCsNPs may be due to their higher cellular uptake as compared to curcumin. Chitosan nanoparticles were not only stable but also a nontoxic. Our data suggested that curcumin loaded nanoformulations, therefore, might be promising candidates in cancer therapy.

Characterization and carboplatin loaded chitosan nanoparticles for the chemotherapy against breast cancer in vitro studies

Aim of the studies to synthesized chitosan nanoparticles by an ionic interaction procedure. The nanoparticles were characterized by physicochemical methods like, DLS, TEM, Surface potential measurements, FT-IR and DSC. The average particle size of chitosan and carboplatin nanoparticles was found to be 277.25±11.37nm and 289.30±8.15nm and zeta potential was found to be 31±3.14mV and 33±2.15mV respectively with low polydispersity index. The maximum entrapment of carboplatin in nanoparticles was a spherical shape with a positive charge. The maximum encapsulation and loading efficiencies of carboplatin (5mg/ml) were obtained to be 58.43% and 13.27% respectively. The nanocarboplatin was better blood compatibility as compared to chitosan nanoparticles. Finally, the cytotoxic effects of the carboplatin loaded chitosan nanoparticles were tested in-vitro against breast cancer (MCF-7) cell lines. Our studies showed that the chitosan nanoparticles could be used as a promising candidate for drug delivery for the therapeutic treatment of breast cancer.

Monocyclic β-lactam and unexpected oxazinone formation: synthesis, crystal structure, docking studies and antibacterial evaluation

Abstract Novel monocyclic β-lactam derivatives bearing aryl, phenyl and heterocyclic rings were synthesized as possible antibacterial agents. Cyclization of imines (3h, 3t) with phenylacetic acid in the presence of phosphoryl chloride and triethyl amine did not afford the expected β-lactams. Instead, highly substituted 1,3-oxazin-4-ones (4h, 4t) were isolated as the only product and confirmed by single crystal X-ray analysis of 4t. The results of antibacterial activity showed that compound 4l exhibited considerable antibacterial activity with MIC and MBC values of 62.5 µg/mL against Klebsiella pneumoniae. Cytotoxicity assay on Chinese Hamster Ovary (CHO) cell line revealed non-cytotoxic behavior of compounds 4d, 4h, 4k and 4l up to 200 μg/mL conc. Molecular docking was performed for compound 4l with penicillin binding protein-5 to identify the nature of interactions. The results of both in silico and in vitro evaluation provide the basis for compound 4l to be carried as a potential lead molecule in the drug discovery pipeline against bacterial infections.

Diketo acids and their amino acid/dipeptidic analogues as promising scaffolds for the development of bacterial methionine aminopeptidase inhibitors

Using diketoesters as the template, various derivatives were designed and the selected compounds were synthesized as bacterial methionine aminopeptidase (MetAP) inhibitors. The results of in vitro antibacterial screening revealed fifteen compounds (1a–c, 1e–h, 1j, 1l, 2a–c, 3d, 5c and 5e) as potent against different bacterial strains. By using the MTT assay on human cell line (HepG2), the viability of cell proliferation was evaluated and nine compounds (1c, 1e, 1j, 1l, 2a,b, 3d, 5c and 5e) showed no cytotoxic effect at the concentration range of 50–450 μg ml−1. In the biochemical evaluation against methionine aminopeptidase (MetAPs) from Streptococcus pneumonia (SpMetAP), Mycobacterium tuberculosis (MtMetAP), Enterococcus faecalis (EfMetAP) and human (HsMetAP), compounds displayed differential behaviour against these four enzymes. Moreover, compound 1g showed 84% inhibition of SpMetAP, while compound 3d selectively inhibited MtMetAP with 79% inhibition and little effect on HsMetAP at 100 μM concentration. At the same concentration, compound 5e exhibited 87% and 85% inhibition of EfMetAP and SpMetAP, respectively. Understanding the mode of binding through modeling at the active site provided the structural basis for the possible mode of inhibition. Together, these data will be useful for further development of diketo acid based inhibitors with improved potency and selectivity.

Ethyl acetate Salix alba leaves extract-loaded chitosan-based hydrogel film for wound dressing applications

High toxicity and multidrug resistance associated with various standard antimicrobial drugs have necessitated search for safer alternatives in plant-derived materials. In this study, we performed biological examination of chitosan-based hydrogel film loaded with ethyl acetate Salix alba leaves extract against 11 standard laboratory strains. FTIR showed regeneration of saccharide peak in CP1A at 1047 cm−1 and increased in height of other peaks. DSC exothermic decomposition peaks at 112 °C, 175 °C and 251 °C reveal the effect of extract on hydrogel film. From FESEM images, three-dimensional cross-linking and extract easily seen in the globular form from the surface. MTT assay on HEK 293 cells showed that CP1A was non-toxic. Minimum inhibitory concentration ranges from 4000 μg/ml to 125 μg/ml. Enterococcus faecium, Candida glabrata and Candida tropicalis were the most resistant, while Salmonella typhi and Candida guilliermondii were the most susceptible micro-organisms.

Phytochemical Composition of Extracts Prepared from Dietary Cucurbitsand their Cytoprotective Efficacies against Hydrogen Peroxide InducedOxidative Stress in GI Cell-INT407

Compositions of phytochemicals in aqueous extracts prepared from Lagenaria siceraria (Ls), Luffa cylindrica (Lc) and Cucurbita maxima (Cm) fruits were identified by gas chromatography and mass spectroscopy. GC-MS analysis showed the presence of 83, 72 and 74 number of phytochemicals in Ls, Lc and Cm respectively. Extracts were evaluated for their role in protection of INT407 cells against injurious effect of H2O2. MTT, LDH and colony survival assays (CSA) were employed for evaluating H2O2-induced cytotoxicity. Antioxidant properties of extracts were evaluated by enzymatic assays in H2O2 induced oxidative stress in INT407 cells. LD50 and LD25 of H2O2 at 24 h on INT407 cells were evaluated and found 114.25 ± 0.24 and 64.01 ± 0.11 μM respectively. Macromolecules were found to protect INT407 in dose dependent manner (p ≤ 0.05). CSA showed enhanced growth in extracts of Lagenaria siceraria and Luffa cylindrica in comparison to Cucurbita maxima treated cells (p=0.042). H2O2 treated cells showed depleted superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and glutathione reductase (GR) activities by 71.60, 68.81, 45.79 and 22.95% respectively. Lagenaria siceraria and Luffa cylindrica treated cells showed modulation of SOD by 8.88 and 8.95%, CAT by 22.34 and 14.20%, GPx by 5.56 and 1.42%, and GR by 2.20 and 3.22% respectively. Cucurbita maxima modulated CAT by 1.78%, and GR by 0.70%. Synergistic effects of phytochemicals present in these extracts were found non-toxic on INT407.

Nutraceuticals as Chemopreventive and Therapeutic agents in Gut AssociatedPathogenesis

Nutraceuticals have gained great insights in recent years due to their therapeutic implications. Secondary metabolites like glutamate, flavonoids, polyphenols and terpenoids are widespread in nature and are part of human diet. Polysaccharides of prebiotic nature are used both as therapeutic and prophylactic agents. These nutraceuticals in recent times have been reported to posses great potential to reduce antigenic and oxidative pressure in the human gut. Antioxidant rich diet is a potential therapeutic agent against reactive oxygen species induced pathogenesis. Epegenetic compounds present in antioxidant and prebiotics rich nutraceuticals mitigate and remove the causative factors associated with pathogenesis. Flavonoids and polyphenols exhibit antioxidant properties and have been recognized as potential gastroprotective agents and epigenetic modulators as well. Triterpenoids such as cucurbitacin is reported to posses anticancer activities. Glutamate is an enteric neurotransmitter that is used in improving neonatal gut function. This review comprehensively summarizes the current knowledge of gut modulating nutraceuticals which ultimately can play its role from prophylaxis of gut to its therapeutics by employing various macro and micro molecular pathways.