A Co(III) Complex of 1-Amino-4-hydroxy-9,10-anthraquinone Exhibits Apoptotic Action against MCF-7 Human Breast Cancer Cells.

A Co(III) complex of 1-amino-4-hydroxy-9,10-anthraquinone (QH) (Scheme-1) having the molecular formula CoQ3 (Scheme-2) was prepared and characterized by elemental analysis, FTIR spectroscopy, UV-vis spectroscopy, fluorescence spectroscopy, and mass spectrometry. In the absence of a single crystal, the energy-optimized molecular structure of CoQ3 was determined by employing computational methods that was validated using spectroscopic evidences, elemental analysis, and mass spectrometry data. The electrochemical properties of the complex were analyzed using cyclic voltammetry and indicate a substantial modification of the electrochemical properties of the parent amino-hydroxy-9,10-anthraquinone. CoQ3 was thereafter tested on MCF-7 human breast cancer cells. The IC50 value for a 24 h incubation was found to be (95 ± 0.05) µg/mL. The study showed that such cancer cells underwent both early and late apoptosis following the interaction with CoQ3.

Bio-functionalized zinc oxide nanoparticles: Potential toxicity impact on freshwater fish Cyprinus carpio.

There is a growing concern nowadays over the exposure of nanomaterials and their effects in aquatic life. In spite of reporting the changes in physiology, reproduction and behaviour in fish by different nanoparticles, the molecular events underlying in the aquatic bodies due to the toxicity of zinc oxide nanoparticles (ZnO NPs) are mainly unexplored. Therefore, the present study carried out an ex vivo exposure of ZnO NPs at various concentrations (0.382, 0.573 and 1.146 mg L-1) in freshwater fish Cyprinus carpio to investigate the potential adverse effects. The results revealed that ZnO NPs exposure altered the haematological parameter and induces the reactive oxygen species (ROS) that leads to elevation of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidise (GPx), glutathione S-transferase (GST) and reduced glutathione (GSH) activity in C. carpio. Furthermore, histopathological analysis exhibited that the ZnO NPs caused lamellar fusion, aneurism, cytoplasmic vacuolation, nuclear alteration, necrotic muscle fiber and pyknotic nuclei in the gills, liver and muscles of C. carpio. ZnO NPs exposure significantly up-regulated the overlapping expressions of SOD1, CAT, GPx1a, GST-α, CYP1A, and Nrf-2 genes. A higher level of Zn bioaccumulation was observed in the following order: gill (35.03 ± 2.50 µg g-1), liver (5.33 ± 0.73 µg g-1) and muscle (2.30 ± 0.20 µg g-1) at 1.146 mg L-1 exposure of ZnO NPs. Hence, the current study indicated that the biogenic ZnO NPs generate toxicity in fishes by modifying the antioxidant defense mechanisms, histomorphology, and oxidative stress encoding genes.

Structural characterization, functional and biological activities of an exopolysaccharide produced by probiotic Bacillus licheniformis AG-06 from Indian polyherbal fermented traditional medicine.

An exopolysaccharide (EPS) was purified from the probiotic bacterium Bacillus licheniformis AG-06 isolated from the polyherbal fermented traditional medicine (Ashwagandharishta) of Indian Ayurveda. High-performance liquid chromatography (HPLC) based compositional analysis exhibits the heteropolymeric nature of the EPS consisting of galactose, rhamnose, xylose, mannose, and glucose, as the monomeric units. Fourier-transform infrared (FT-IR) and nuclear magnetic resonance (NMR) spectroscopic analyses confirm the presence of typical carbohydrate polymer functional groups and structural units, respectively. The purified EPS demonstrates the web-like fibrous and porous nature in scanning electron microscopic and atomic force microscopic studies. The purified EPS had shown 71.83% and 67.79% of flocculation and emulsification activities, respectively. Antioxidant activity was evaluated against 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), nitric oxide, and superoxide free radicals and the scavenging actions were increased in a dose-dependent manner. Moreover, the purified EPS exhibits a significant cytotoxic activity against the human lung carcinoma cells (A549), which strongly suggests the anticancer potential of the EPS derived from B. licheniformis AG-06.

Characterization of Hen's Egg White To Use It as a Novel Platform To Culture Three-Dimensional Multicellular Tumor Spheroids.

We are standardizing protocols to develop egg white (EW) as a cost-effective platform for culture of three-dimensional (3-D) multicellular tumor spheroids for application in understanding tumor microenvironments and drug screening. In this article, we describe several physical and physiological characteristics of EW to use it as 3-D cell culture platform. Field emission scanning electron microscopy revealed the presence of different microstructures. Hydrodynamic size distribution data indicated nano- and micron-sized particles. Rheological measurements revealed the viscosity and viscoelastic behavior appropriate for maintaining cell viability and supporting 3-D cell growth under high-sheer conditions. It was found that thereis no autofluorescence, a requirement for imparting transparency and for microscopic observations of the spheroids. The EW facilitated the development of 3-D tumor spheroids, with an emphasis of difference in cell proliferation and intercellular cytoskeletal organization between two-dimensional and 3-D spheroid cultures. Put together, EW proves to be a cost-affordable and simple platform for 3-D culture of tumor spheroids.

Co-delivery of Diverse Therapeutic Compounds Using PEG-PLGA Nanoparticle Cargo against Drug-Resistant Bacteria: An Improved Anti-biofilm Strategy.

Controlling biofilms of bacteria is a challenging aspect because of their drug-resistance potentials against a range of antibiotics, demanding the development of active anti-biofilm agents. Rutin (R), a natural antioxidant, and benzamide (B), a synthetic antibacterial agent, have several pharmacological and antibacterial abilities. Herein, we developed PEG-PLGA NPs that synergistically carried rutin and benzamide as drug candidates, while displaying therapeutic and anti-biofilm  functions. These drug delivery NPs were synthesized by the oil-in-water emulsion (O/W) solvent evaporation technique. The obtained NPs were characterized by UV-vis, FT-IR, SEM, TEM, and DLS measurements. Confocal laser scanning microscopy was employed to evaluate the anti-biofilm capabilities against Staphylococcus aureus and Pseudomonas aeruginosa and further quantified the levels of residual biofilm constituents such as protein and exopolysaccharide (EPS). Drug release experiments showed the controlled release of rutin-benzamide (RB) for several days. Antibacterial analyses showed that the minimum inhibitory concentration (MIC) of NPs was at least two times lower than that of the free drugs. RB-PEG-PLGA NPs revealed that they targeted biofilm-forming bacteria through the disruption of the membrane and biofilm surface and were observed to be nontoxic when tested using human erythrocytes and human cell lines. In vivo evaluations in zebrafish showed that the NPs did not alter the antioxidant functions and histological features of tissues. On the basis of results obtained, it is substantiated that the rutin-benzamide-loaded nanocarrier offers potential anti-biofilm therapy due to its high anti-biofilm activity and biocompatibility.

Dual drug loaded PLGA nanospheres for synergistic efficacy in breast cancer therapy.

Improved therapeutic effects can be achieved by the delivery of combination of drugs through multifunctional cell targeted nanocarrier systems. The present investigation reports the preparation of Poly (D,L-lactic-co-glycolic acid) (PLGA) nanospheres loaded with the novel combination such as Rutin (R) and Benzamide (B) as drugs using water-oil-water (w/o/w) emulsion method. Dual drug loaded PLGA nanospheres (R/B@PLGA) were stabilized by poly (vinyl alcohol) (PVA) coating and characterized in terms of morphology, size, surface charge, and structural chemistry by Scanning electron microscopy (SEM), Dynamic light scattering (DLS), Zeta potential analysis, UV-vis and Fourier transform infrared (FT-IR) spectroscopy. The inhibitory effects of rutin and benzamide on MDA-MB-231 (triple negative breast cancer-TNBC) cells using the drug loaded PLGA nanospheres as well as their non-toxic features were evaluated in vivo. The anticancer activity of the R/B@PLGA nanospheres through cell cycle disruption and apoptotic induction was assessed in vitro by flow cytometry analysis. Further, the in vitro antioxidant capacity, pH-based drug release and hemocompatible property were also investigated. It was shown that the R/B@PLGA nanospheres lacked genotoxic potential and they did not alter the antioxidant enzyme activities and histological features of zebrafish. Hence, this dual drug delivery system (DDS) not only actively targets multidrug-resistance (MDR) associated phenotype but also improves the therapeutic efficiency by its non-toxic nature towards enhanced cancer cell focused delivery and sustained release of therapeutic agents.

Antibacterial synergy between rutin and florfenicol enhances therapeutic spectrum against drug resistant Aeromonas hydrophila.

Emergence of antibiotic resistant bacteria has necessitated the drive to explore competent antimicrobial agents or to develop novel formulations to treat infections including Aeromonas hydrophila. The present study investigates the synergistic antibacterial effects of citrus flavonoid rutin and florfenicol (FF) against A. hydrophila in vitro and in vivo. Rutin is extracted and purified from Citrus sinensis peel through preparative HPLC and characterized through TLC, GC-MS and 1H and 13C NMR analyses. Though rutin did not display significant antibacterial activity, it modulated FF activity resulting in four-fold reduction in the MIC value for FF. The anti-biofilm potential of synergistic association of rutin and FF was validated by protein analysis, quantification of exopolysaccharide (EPS) and microscopy studies using sub-MIC doses. Besides antibacterial action, in vivo studies showed that Rutin/FF combination enhanced host immunity by improving blood cell count, anti-protease, and lysozyme activities as well as decreased the oxidative stress and the pathological changes of tilapia Oreochromis niloticus against A. hydrophila infection. No significant DNA damages or clastogenic effects were detected in tilapia challenged with A. hydrophila under Rutin/FF treatment. It is shown that an acute-phase Lipopolysaccharide binding protein (LBP) enhances the innate host defence against bacterial challenge. Semi quantitative RT-PCR and western blot results revealed the significant increase of LBP in the supernatant of tilapia monocytes/macrophages challenged with A. hydrophila upon treatment. The study findings substantiate that the combination of natural molecules with antibiotics may open up possibilities to treat MDR strains.

A novel rutin-fucoidan complex based phytotherapy for cervical cancer through achieving enhanced bioavailability and cancer cell apoptosis.

Recent studies on flavonoids forming complexes with macromolecules attract researchers due to their enhanced bioavailability as well as chemo-preventive efficacy. In this study, a flavonoid rutin (Ru) is non-covalently complexed with fucoidan (Fu) using the functional groups to obtain a therapeutic polymeric complex overcoming the limitations of bioavailability of rutin. The prepared novel rutin-fucoidan (Ru-Fu) complex is characterized for spectroscopic features, particle size and distribution analysis by DLS. It is shown that the complex displayed the nanostructural features that are different from that of the usual rutin-fucoidan mixture. The studies on drug release profiles at different pH (5.5, 6.8 and 7.4) show that the sustained release of compounds from complex occurs preferentially at the desired endosomal pH (5.5). Further, the chemopreventive potential of Ru-Fu complex is investigated against HeLa cells by cellular apoptotic assays and flow cytometric analysis. It showed that the complex is able to disrupt cell cycle regulation and has the ability to induce cellular apoptosis via nuclear fragmentation, ROS generation and mitochondrial potential loss. In vitro cell viability assay with Ru-Fu complex shows that the complex is biocompatible on normal cells. The hemolysis assay also reveals that the complex does not release hemoglobin from human red blood cells (RBCs). Thus, the study is envisaged to open up interests for developing such formulations against cervical cancer and other cancers.

Combinatorial nanocarrier based drug delivery approach for amalgamation of anti-tumor agents in breast cancer cells: an improved nanomedicine strategy.

Combination therapy of multiple drugs through a single system is exhibiting high therapeutic effects. We investigate nanocarrier mediated inhibitory effects of topotecan (TPT) and quercetin (QT) on triple negative breast cancer (TNBC) (MDA-MB-231) and multi drug resistant (MDR) type breast cancer cells (MCF-7) with respect to cellular uptake efficiency and therapeutic mechanisms as in vitro and in vivo. The synthesized mesoporous silica nanoparticle (MSN) pores used for loading TPT; the outer of the nanoparticles was decorated with poly (acrylic acid) (PAA)-Chitosan (CS) as anionic inner-cationic outer layer respectively and conjugated with QT. Subsequently, grafting of arginine-glycine-aspartic acid (cRGD) peptide on the surface of nanocarrier (CPMSN) thwarted the uptake by normal cells, but facilitated their uptake in cancer cells through integrin receptor mediated endocytosis and the dissociation of nanocarriers due to the ability to degrade of CS and PAA in acidic pH, which enhance the intracellular release of drugs. Subsequently, the released drugs induce remarkable molecular activation as well as structural changes in tumor cell endoplasmic reticulum, nucleus and mitochondria that can trigger cell death. The valuable CPMSNs may open up new avenues in developing targeted therapeutic strategies to treat cancer through serving as an effective drug delivery podium.

Immune response and disease resistance of carotenoids supplementation diet in Cyprinus carpio against Aeromonas hydrophila.

The effect of carotenoid-supplementation diet on immune response and disease resistance in common carp, Cyprinus carpio against Aeromonas hydrophila at weeks 1, 2, and 4 is reported. The cumulative mortality was 10% when fish were fed with 50 or 100 mg kg(-1) supplementation diets while the un-supplementation diet treated group suffered 90% mortality against the pathogen. The phagocytic activity and complement activity significantly increased with 50 and 100 mg kg(-1) diet groups from weeks 2 and 4 but not in other groups. The reactive oxygen species (ROS) production was significantly enhanced with 50 and 100 mg kg(-1) diets from weeks 1 to 4 while the production of reactive nitrogen species (RNS) enhanced on weeks 2 and 4. The lysozyme activity significantly increased when fed with 50 and 100 mg kg(-1) diets on weeks 2 and all supplementation diets on week 4. These results suggest that diet enriched with carotenoid pigment positively enhance the immune status and protects C. carpio from A. hydrophila infection.

Protective effect of chitin and chitosan enriched diets on immunity and disease resistance in Cirrhina mrigala against Aphanomyces invadans.

The effect of diet enriched with 1% chitin or chitosan on innate immune response and disease resistance in Cirrhina mrigala against Aphanomyces invadans was investigated at weeks 1, 2, and 4. In the un-infected and infected groups the white blood cells (WBC), red blood cells (RBC), haematocrit (Ht), lympocytes, monocytes, and neutrophils significantly increased when fed with 1% chitin (CH) or chitosan (CT) enriched diet from weeks 1 to 4 when compared to control; however, the haemoglobin (Hb) and thrombocytes significantly increased only on weeks 2 and 4. The total protein and albumin levels also significantly increased with any enriched diet on weeks 2 and 4; but the globulin and albumin:globulin ratio increased on week 4 as compared to control; similarly the phagocytic activity significantly increased on weeks 2 and 4 while the lysozyme activity increased from weeks 1 to 4. The complement activity was significantly enhanced in CT and CTI fed groups on weeks 2 and 4. In un-infected fish fed with 1% CH and CT diets, the cumulative mortality was 10% and 5% whereas the infected fish suffered 20% and 25% mortality. The present results suggest that infected fish fed with 1% chitin or chitosan enriched diet modulates the immune system conferring disease resistance in C. mrigala against A. invadans.

Activation of intrinsic apoptotic signaling pathway in cancer cells by Cymbopogon citratus polysaccharide fractions.

Essential oils of Cymbopogon citratus were already reported to have wide ranging medical and industrial applications. However, information on polysaccharides from the plant and their anticancer activities are limited. In the present study, polysaccharides from C. citratus were extracted and fractionated by anion exchange and gel filtration chromatography. Two different polysaccharide fractions such as F1 and F2 were obtained, and these fractions were found to have distinct acidic polysaccharides as characterized by their molecular weight and sugar content. NMR spectral analysis revealed the presence of (1→4) linked b-d-Xylofuranose moiety in these polysaccharides. Using these polysaccharide fractions F1 and F2, anti-inflammatory and anticancer activities were evaluated against cancer cells in vitro and the mechanism of action of the polysaccharides in inducing apoptosis in cancer cells via intrinsic pathway was also proposed. Two different reproductive cancer cells such as Siha and LNCap were employed for in vitro studies on cytotoxicity, induction of apoptosis and apoptotic DNA fragmentation, changes in mitochondrial membrane potential, and profiles of gene and protein expression in response to treatment of cells by the polysaccharide fractions. These polysaccharide fractions exhibited potential cytotoxic and apoptotic effects on carcinoma cells, and they induced apoptosis in these cells through the events of up-regulation of caspase 3, down-regulation of bcl-2 family genes followed by cytochrome c release.

Impact of tobacco on glutathione S transferase gene loci of Indian ethnics.

BACKGROUND: Tobacco contains agents which generate various potent DNA adducts that can cause gene mutations. Production of DNA adducts may be neutralized by glutathione S transferase (GST) along with other phase I and phase II enzyme systems. The existence of null type of GST among the population increases the susceptibility to various disorders and diseases. The present study focuses on the impact of high tobacco usage and possible null type mutation in GST loci. METHODS: Genotypes of GST were detected by multiplex polymerase chain reaction in unrelated 504 volunteers of high tobacco using natives of Gujarat. Allelic frequencies were calculated using Statistical Package for Social Studies-16 software. Hardy Weinberg Equilibrium (HWE) was calculated using Chi square test. Two sided Fisher's significance test was used to compare allelic frequencies of different populations. RESULTS: The frequency of homozygous null genotype of GSTM1 and GSTT1 were 20% (95% CI 16.7-23.9) and 35.5% (95% CI 31.4-39.9) respectively. The GSTM1 and GSTT1 null allele frequency distribution in the Gujarat population was significantly deviating from HWE. GSTT1 null frequency of Gujaratians was significantly higher and different to all reported low tobacco using Indian ethnics, while GSTM1 was not differing significantly. CONCLUSION: Tobacco usage significantly influences the rate of mutation and frequency of GSTT1 and M1 null types among the habituates. The rate of mutation in GSTT1 loci was an undeviating response to the dose of tobacco usage among the population. This mutational impact of tobacco on GSTT1 postulates the possible gene - environment interaction and selection of null genotype among the subjects to prone them under susceptible status for various cancers and even worst to cure the population with GSTT1 dependent drugs.

Survivin deficiency leads to imparalization of cytokinesis in cancer cells.

BACKGROUND: Survivin has been implicated in cancer progression and is known to be over-expressed in a variety of human malignancies. Positive regulation of survivin expression provides a connecting link between cell cycle and tumorigenesis or perhaps tumour maintenance. METHODS: An experiment was designed to analyse survivin expression in cell lines (MCF 7, Zr751, A549, HepG2) using SDS-PAGE, Western blots, RT-PCR, AGE and heamatoxylin-eosin staining were done. RESULTS: SDS-PAGE revealed the presence of 16.5 kDa protein. Subsequent western Blot and cytological analysis showed down-regulation of survivin expression in cancer cells. CONCLUSION: Therefore, the study allows the conclusion that survivin is essential for proper chromosome segregation and cytokinesis. It seems reasonable to suspect that abnormal expression or function of survivin might contribute to multinucleated and apoptotic conditions.