Braj Raj Singh

Production of antimicrobial silver nanoparticles in water extracts of the fungus Amylomyces rouxii strain KSU-09.

A fungal strain, KSU-09, isolated from the roots of date palm (Phoenix dactylifera), was identified as Amylomyces rouxii based on sequence analysis of the internal transcribed spacer (ITS) region of its rRNA genes. Mycelia-free water extracts obtained from mycelium suspended in water for 72h facilitated the production of stable, predominantly monodispersed and spherical silver nanoparticles (AgNPs) in the size range of 5-27nm upon addition of 1mM silver nitrate, as determined by the XRD, AFM and TEM. The AgNPs exhibited antimicrobial activity against Shigella dysenteriae type I, Staphylococcus aureus, Citrobacter sp., Escherichia coli, Pseudomonas aeruginosa, Bacillus subtilis, Candida albicans and Fusarium oxysporum. Thus, the strain KSU-09 could be used for simple, non-hazardous and efficient synthesis of antimicrobial AgNPs.

Potential chemoprevention of N-nitrosodiethylamine-induced hepatocarcinogenesis by polyphenolics from Acacia nilotica bark

Chemopreventive potential of Acacia nilotica bark extract (ANBE) against single intraperitoneal injection of N-nitrosodiethylamine (NDEA, 200mg/kg) followed by weekly subcutaneous injections of carbon tetrachloride (CCl(4), 3 ml/kg) for 6 weeks induced hepatocellular carcinoma (HCC) in rats was studied. At 45 day after administration of NDEA, 100 and 200mg/kg of ANBE were administered orally once daily for 10 weeks. The levels of liver injury and liver cancer markers such as alanine transaminase (ALT), aspartate transaminase (AST), alkaline phosphatase (ALP), gamma-glutamyl transferase (gamma-GT), total bilirubin level (TBL), alpha-feto protein (AFP) and carcinoembryonic antigen (CEA) were substantially increased following NDEA treatment. However, ANBE treatment reduced liver injury and restored liver cancer markers. ANBE also significantly prevented hepatic malondialdehyde (MDA) formation and reduced glutathione (GSH) in NDEA-treated rats which was dose dependent. Additionally, ANBE also increased the activities of antioxidant enzymes viz., catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPx), and glutathione-S-transferase (GST) in the liver of NDEA-administered rats. Eventually, ANBE also significantly improved body weight and prevented increase of relative liver weight due to NDEA treatment. Histological observations of liver tissues too correlated with the biochemical observations. HPLC analysis of ANBE showed the presence of gallic, protocatechuic, caffeic and ellagic acids, and also quercetin in ANBE. The results strongly support that A. nilotica bark prevents lipid peroxidation (LPO) and promote the enzymatic and non-enzymatic antioxidant defense system during NDEA-induced hepatocarcinogenesis which might be due to activities like scavenging of oxy radicals by the phytomolecules in ANBE.

ROS-dependent anticandidal activity of zinc oxide nanoparticles synthesized by using egg albumen as a biotemplate

Zinc oxide nanoparticles (ZnO NPs) have attracted great attention because of their superior optical properties and wide application in biomedical science. However, little is known about the anticandidal activity of ZnO NPs against Candida albicans (C. albicans). This study was designed to develop the green approach to synthesize ZnO NPs using egg white (denoted as EtZnO NPs) and investigated its possible mechanism of antimicrobial activity against C. albicans 077. It was also notable that anticandidal activity of EtZnO NPs is correlated with reactive oxygen species (ROS) production in a dose dependent manner. Protection of histidine against ROS clearly suggests the implication of ROS in anticandidal activity of EtZnO NPs. This green approach based on egg white-mediated synthesis of ZnO NPs paves the way for developing cost effective, eco-friendly and promising antimicrobial nanomaterial for applications in medicine.

Biofabricated Silver Nanoparticles Act as a Strong Fungicide against Bipolaris sorokiniana Causing Spot Blotch Disease in Wheat

The present study is focused on the extracellular synthesis of silver nanoparticles (AgNPs) using culture supernatant of an agriculturally important bacterium, Serratia sp. BHU-S4 and demonstrates its effective application for the management of spot blotch disease in wheat. The biosynthesis of AgNPs by Serratia sp. BHU-S4 (denoted as bsAgNPs) was monitored by UV–visible spectrum that showed the surface plasmon resonance (SPR) peak at 410 nm, an important characteristic of AgNPs. Furthermore, the structural, morphological, elemental, functional and thermal characterization of bsAgNPs was carried out using the X-ray diffraction (XRD), electron and atomic microscopies, energy dispersive X-ray (EDAX) spectrometer, FTIR spectroscopy and thermogravimetric analyzer (TGA), respectively. The bsAgNPs were spherical in shape with size range of ∼10 to 20 nm. The XRD and EDAX analysis confirmed successful biosynthesis and crystalline nature of AgNPs. The bsAgNPs exhibited strong antifungal activity against Bipolaris sorokiniana, the spot blotch pathogen of wheat. Interestingly, 2, 4 and 10 µg/ml concentrations of bsAgNPs accounted for complete inhibition of conidial germination, whereas in the absence of bsAgNPs, conidial germination was 100%. A detached leaf bioassay revealed prominent conidial germination on wheat leaves infected with B. sorokiniana conidial suspension alone, while the germination of conidia was totally inhibited when the leaves were treated with bsAgNPs. The results were further authenticated under green house conditions, where application of bsAgNPs significantly reduced B. sorokiniana infection in wheat plants. Histochemical staining revealed a significant role of bsAgNPs treatment in inducing lignin deposition in vascular bundles. In summary, our findings represent the efficient application of bsAgNPs in plant disease management, indicating the exciting possibilities of nanofungicide employing agriculturally important bacteria.

Synthesis of stable cadmium sulfide nanoparticles using surfactin produced by Bacillus amyloliquifaciens strain KSU-109

In this study, a surfactin was extracted from a novel surfactant producing bacterial strain Bacillus amyloliquifaciens KSU-109, isolated from rhizosphere of date palm (Phoenix dactylifera), and characterized based on 16Sr RNA and sfp genes using Blastn, Blastx and phylogenetic analyses. The study was performed to obtain a renewable bioresource for surfactin production, and its application in nanotechnology as a non-hazardous and environmentally compatible nanoparticle (NP) stabilizer. The strain KSU-109 produced the surfactin with an average yield of 160 mg/L with strong surfactant activity, reducing the surface tension of the medium from 72 mN/m to 29.3 mN/m. The surfactin preparation was used for synthesizing the cadmium sulfide nanoparticles (CdS-NPs) by mixing 0.005% surfactin with 1mM Cd(NO(3))(2) in 1:1 ratio (v/v) and 10mM Na(2)S solution at pH 7.2 and ambient temperature, which were stable up to 120 days. The surfactin stabilized CdS-NPs were characterized using XRD, TEM, and spectroscopic techniques. The data revealed a significant role of surfactin as a stabilizer and capping agent, which also causes phase transition to yield the cubic/hexagonal CdS-NPs of average size of 3-4 nm. The results elucidated the significance of biocompatible and biodegradable surfactin as an effective and inexpensive stabilizing agent for developing stable CdS nanoparticles.

Biosynthesis of Stable Antioxidant ZnO Nanoparticles by Pseudomonas aeruginosa Rhamnolipids

During the last several years, various chemical methods have been used for synthesis of a variety of metal nanoparticles. Most of these methods pose severe environmental problems and biological risks; therefore the present study reports a biological route for synthesis of zinc oxide nanoparticles using Pseudomonas aeruginosa rhamnolipids (RLs) (denoted as RL@ZnO) and their antioxidant property. Formation of stable RL@ZnO nanoparticles gave mostly spherical particles with a particle size ranging from 35 to 80 nm. The RL@ZnO nanoparticles were characterized by UV-visible (UV–vis) spectroscopy, scanning electron microscopy, transmission electron microscopy, dynamic light scattering, Fourier transform infrared spectroscopy, X-ray diffraction (XRD), and thermal gravimetric analysis. The UV–vis spectra presented a characteristic absorbance peak at ∼360 nm for synthesized RL@ZnO nanoparticles. The XRD spectrum showed that RL@ZnO nanoparticles are crystalline in nature and have typical wurtzite type polycrystals. Antioxidant potential of RL@ZnO nanoparticles was assessed through 2,2–diphenyl-1-picrylhydrazyl (DPPH), hydroxyl, and superoxide anion free radicals with varying concentration and time of the storage up to 15 months, while it was found to decline in bare ZnO nanoparticles. Similarly, the inhibitory effects on β-carotene oxidation and lipid peroxidation were also observed. These results elucidate the significance of P. aeruginosa RL as effective stabilizing agents to develop surface protective ZnO nanoparticles, which can be used as promising antioxidants in biological system.

Mycofabricated biosilver nanoparticles interrupt Pseudomonas aeruginosa quorum sensing systems.

Quorum sensing (QS) is a chemical communication process that Pseudomonas aeruginosa uses to regulate virulence and biofilm formation. Disabling of QS is an emerging approach for combating its pathogenicity. Silver nanoparticles (AgNPs) have been widely applied as antimicrobial agents against human pathogenic bacteria and fungi, but not for the attenuation of bacterial QS. Here we mycofabricated AgNPs (mfAgNPs) using metabolites of soil fungus Rhizopus arrhizus BRS-07 and tested their effect on QS-regulated virulence and biofilm formation of P. aeruginosa. Transcriptional studies demonstrated that mfAgNPs reduced the levels of LasIR-RhlIR. Treatment of mfAgNPs inhibited biofilm formation, production of several virulence factors (e.g. LasA protease, LasB elastrase, pyocyanin, pyoverdin, pyochelin, rhamnolipid, and alginate) and reduced AHLs production. Further genes quantification analyses revealed that mfAgNPs significantly down-regulated QS-regulated genes, specifically those encoded to the secretion of virulence factors. The results clearly indicated the anti-virulence property of mfAgNPs by inhibiting P. aeruginosa QS signaling.

The vibrational spectra of SO2−4 ion in the hydrated double sulphates: The tutton salts

Abstract The infrared spectra of ten double sulphates, viz., M′ 2 M″(SO 4 ) 2 6H 2 O (M′  NH 4 /K; M″  Ni, Co, Fe, Cu, Zn and Mn) have been recorded in the 1300–400 cm −1 region and the vibrational spectra of SO 2− 4 ion in these salts compared. Spectroscopic observations are correlated with the known SO bond lengths in these salts. The spectrum of the KCu salt is distinct from that of the others and the existence of two types of SO 2− 4 ions in this salt is suggested. In all the salts, the SO 2− 4 ion exhibits linear as well as angular distortion from its free-state tetrahedral symmetry. The linear distortion is, however, greatest in the Mn salt, consistent with known crystal structure data.

Evaluation of the Toxic Potential of Graphene Copper Nanocomposite (GCNC) in the Third Instar Larvae of Transgenic Drosophila melanogaster (hsp70-lacZ)Bg9

Graphene, a two-dimensional carbon sheet with single-atom thickness, have attracted the scientific world for its potential applications in various field including the biomedical areas. In the present study the graphene copper nanocomposite (GCNC) was synthesized, characterized and evaluated for its toxic potential on third instar larvae of transgenic Drosophila melanogaster (hsp70-lacZ)Bg9. The synthesized GCNC was analyzed by X-ray diffraction (XRD), scanning/transmission electron microscopy (SEM/TEM), atomic force microscopy (AFM), and fourier transform infrared spectroscopy (FTIR). The GCNC in 0.1% DMSO was sonicated for 10 min and the final concentration of 0.033, 0.099, 0.199 and 3.996 µg/µl of diet were established. The third instar larvae were allowed to feed on it separately for 24 and 48 hrs. The hsp70 expression was measured by O-nitrophenyl-β-D-galactopyranoside assay, tissue damage by trypan blue exclusion test and β-galactosidase activity was monitored by in situ histochemical β-galactosidase staining. Oxidative stress was monitored by performing lipid peroxidation assay and total protein estimation. Ethidium bromide/acridine orange staining was performed on midgut cells for apoptotic index and the comet assay was performed for the DNA damage. The results of the present study showed that the exposure of 0.199 and 3.996 µg/µl of GCNC were toxic for 24 hr of exposure and for 48 hr of exposure: 0.099, 0.199 and 3.996 µg/µl of GCNC was toxic. The dose of 0.033 µg/µl of GCNC showed no toxic effects on its exposure to the third instar larvae for 24 hr as well as 48 hrs. This dose can be considered as No Observed Adverse Effect Level (NOAEL).

Dietary phytochemicals alter epigenetic events and signaling pathways for inhibition of metastasis cascade

Cancer metastasis is a multistep process in which a cancer cell spreads from the site of the primary lesion, passes through the circulatory system, and establishes a secondary tumor at a new nonadjacent organ or part. Inhibition of cancer progression by dietary phytochemicals (DPs) offers significant promise for reducing the incidence and mortality of cancer. Consumption of DPs in the diet has been linked to a decrease in the rate of metastatic cancer in a number of preclinical animal models and human epidemiological studies. DPs have been reported to modulate the numerous biological events including epigenetic events (noncoding micro-RNAs, histone modification, and DNA methylation) and multiple signaling transduction pathways (Wnt/β-catenin, Notch, Sonic hedgehog, COX-2, EGFR, MAPK-ERK, JAK-STAT, Akt/PI3K/mTOR, NF-κB, AP-1, etc.), which can play a key role in regulation of metastasis cascade. Extensive studies have also been performed to determine the molecular mechanisms underlying antimetastatic activity of DPs, with results indicating that these DPs have significant inhibitory activity at nearly every step of the metastatic cascade. DPs have anticancer effects by inducing apoptosis and by inhibiting cell growth, migration, invasion, and angiogenesis. Growing evidence has also shown that these natural agents potentiate the efficacy of chemotherapy and radiotherapy through the regulation of multiple signaling pathways. In this review, we discuss the variety of molecular mechanisms by which DPs regulate metastatic cascade and highlight the potentials of these DPs as promising therapeutic inhibitors of cancer.