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Abstract: Laccase (Benzenediol: oxygen oxidoreductase; E.C.1.10.3.2), a multicopper oxidase that is a known lignin-degrading enzyme, can catalyse an ample array of substrates, from phenolic, nonphenolic compounds, aromatic amines, diamines, heterocyclic compounds to organic/inorganic metal compounds, etc., bestowed they have not too high redox potentials. Despite many laccase-producing organisms like bacteria, insects, plants, and animals, white rot filamentous fungi are the best producers of this enzyme. In the presence of laccase, pesticides (fungicides, herbicides, insecticides, etc.) of various chemical compositions (organophosphates, organochlorines, carbamates, pyrethrin & pyrethroids, etc.) are oxidized into the water with collateral reduction of four electrons of molecular oxygen with various efficiencies. Bioremediation efficiency can be increased in the presence of various natural or synthetic mediators, viz. ABTS, violuric acid, 1- hydroxy benzotriazole, vanillin, syringaldehyde, PEG, etc. Immobilized laccase on various supporting materials increased the enzyme's stability, reliability, and reusability for continuous application, particularly for industrial processes. The present review discusses the structure, catalytic cycle, general mechanism of oxidation, and various scopes and challenges of pesticide degradation by this multifaceted biocatalyst which could lead to a green sustainable environment.

It is difficult to calculate fixed data of a number of cells in a body. It is varying on time and age of life. The time increasing that cells are increasing, though we can estimate the number of cells in various nerves in a body, brain, sexual platform, etc through a couple system. The coupling system is a new system of the finding of peculiar series of numbers [1], which is applies to many fields. In the cases of Medical Science, it has been observed by calculation that due to disturbing of the couple caused different difficulties in the body. A smooth Coupling cell may produce a healthy body, and it is possible to increase memory by adding particular cells number in a loss position. The coupling system is interesting that, can explain the real mechanism of every cell. There are many types of cells in a living body. Almost all cells follow a couple of system. The coupling system performs coupling between two (say, A, 1st party & B, 2nd party) with keeping relation as 3rd party, denoted by R (Relative Number) mathematically.

Light is very sensitive matter. In terms of mass of a photon is important in every field of matter thus for the universe. The scientists of many countries are trying to find the mass of photon by experiment since 1936 and continuing this work in various countries. But the obtained results are differing to each other. So, we cannot consider these mass of a photon. On the other hand matter is made by the photons. We get this idea from the Einstein equation E = mc2. Again, energy is nothing but the bunch of photons. I calculate the mass of a photon (1.6596x10-54 gm) [1] and this mass is applicable to all fields. Here, in this article, we can calculate the mass of “Curie particle” (Unknown to us) by using this mass of photon & is related to Higgs and other sub-atomic particles. The energy of Higgs particle is very low as per BICEP2`s experimental report [2] and I reported about this matter in the year 2011 to “The Authority of CERN, Editor of Press release of CERN and many other places” by emailing, but did not get answer in this regards. But this is very interesting that as per “Universe Today” report (20th November, 2014) of BICEP2 (Background Imaging of Cosmic Extragalactic Polarization), my calculated observation supports their views. Dr. A. P. J. ABDUL KALAM, SRIJAN PAL SINGH reported (17 June, 2015) the mass neutrino as 1x10-37 kg [3] (in terms of energy is 0.056095861 eV) obtained from calculated results from the difference of two particles. Again, as per report, 19th June, & 27th July 2015, my calculated values tallied “The LHCb collaboration” [4], “ATLAS” [5] experiments. From this view, we have to think on photon`s mass and its activities. From the calculated results, we can say, the mass of a photon is correct and it is applicable to all fields from the particle to the universe.

Disha A (Bacillus cereus) and Disha B (Bacillus safensis) were isolated from pesticide-infested agricultural field and showed tolerance against pesticides, heavy metals, and antibiotics. The isolates exhibited PGPR activities in vitro as well as in field conditions in lentil (Lens culinaris) and cow pea (Vigna unguiculata). Both the Bacillus species could not be grown in mineral salt medium but efficiently grown in the media supplemented with pesticide (imidacloprid/carbendazim) demonstrating the utilization of pesticide as carbon/nitrogen source. The HPLC studies exhibited the pesticide (imidacloprid/carbendazim) degradation by both the bacteria. B. safensis showed better degradation of carbendazim (88.93%) and imidacloprid (82.48%) than that of B. cereus 78.07% and 49.12%, respectively. The bacterial isolates showed high concentration of heavy metal tolerance viz. lead, molybdenum, cadmium, copper, cobalt, and zinc, except mercury. Both the bacteria possessed single plasmid. The plasmid-cured isolates of B. cereus did not tolerate any pesticide, whereas that of B. safensis tolerated all the pesticides, like wild strains. The plasmid curing experiments did not affect the heavy metal tolerance ability of both the bacteria indicating the genomic nature of heavy metal tolerance genes, whereas pesticide resistance genes are plasmid-dependent in B. cereus but genomic in B. safensis.

The area of material synthesis is a rapidly developing field of research which enables scientists not only to discover the high yield, cost effective or environment friendly methods , but also providing them some new opportunities to work in the world of nano science. The green approaches in one hand are the alternative way to synthesize the material by minimizing the wastes as well as toxic substances, on the other hand the synthesis of nanoparticles has gained tremendous attention owing to their many fold applications in various fields. This book is designed to give the readers an outline of some of the very special topics from current prospective of different approaches for material synthesis.

Antagonistic bacteria are common soil inhabitants with potential to control several soil-borne diseases of various crops. In this study, two methomyl degrading Bacillus sp. were screened for their antagonistic potential against soil borne pathogen identified as Alternaria sp. which causes leaf spot and blight disease in lentil. Both the strains produced non-volatile and volatile organic compounds, extracellular enzymes, siderophore, indole acetic acid and solubilized phosphate which ascribed to the mechanism of bio-control and plant growth promotion. These bacterial strains produced indole acetic acid, chitinase and solubilized phosphate even in presence of pesticides (namely methomyl, carbendazim and imidacloprid). The production of chitinase increased by 51–140% in presence of different tested pesticides by the bacterial strains. However, phosphate solublization was only increased up to 79% in B. cereus and 87% in B. safensis in presence of methomyl. Both strains promoted plant growth and suppressed leaf spot and the incidence of blight in lentil plants under controlled conditions in green house. Application of B. cereus and B. safensis isolates to sterile rhizospheric soil increased the dry weight of plants by 40.8% and 43.2%, respectively as compared to control. In similar set of experiments the disease incidence was reduced by 67.7–81.6% in B. cereus and 57.2–78.8% in B. safensis in sterile condition and by 51.4–76.5% and 48.6–63.4%, respectively in non-sterile condition. The present investigation shows both B. cereus and B. safensis as potential plant growth promoting rhizobacteria that can be exploited as efficient bio-control organisms against soil borne plant pathogens as well as can be applied in plant growth enhancement even in pesticide infested soil.

Cerebral ischemia-reperfusion (CIR) injury occurs as a result of oxygen occlusion in the carotid artery through embolus or thrombus formation or cerebrovascular hemorrhage. The oxygen thrust during reperfusion causes the generation of reactive oxidative species (ROS) which exert a potential threat to neuronal survival. ROS may possibly be arrested by antioxidants. After CIR, extracellular matrix remodeling takes place, which is governed by matrix metalloproteinases (MMPs). Augmentation of lipid per oxidation, perturbation of antioxidant enzyme activities and the loss of pyramidal neuronal cells in rat brain were attributed to CIR injury. Melatonin can readily cross the blood-brain barrier (BBB) to exert protective effects as an antioxidant but it is quickly cleared by the circulating blood. Also melatonin is easily degraded by light and hence is found to be ineffective during daytime. Results of the present study showed that unlike free melatonin (FM), the application of nanocapsulated melatonin (NM) exhibited significantly higher potential even at much lower concentrations to rescue neuronal cells and mitochondria during CIR insult and also restored the activities of antioxidative enzymes and MMPs to their normal levels. Hence, nanoencapsulated melatonin may be considered as a suitable drug delivery system for brain to exert protection against CIR injury.

Suvomoy Sana, Swarupa Ghosh, Nirmalendu Das, Sibani Sarkar, Ardhendu Kumar Mandal Drug Development, Diagnostics and Biotechnology, CSIR-Indian Institute of Chemical Biology, West Bengal, India Abstract: The importance of fluoride as a natural and industrial toxicant is recognized worldwide. We evaluated the regulating role and biological effect of vesicular (liposomal and nanoencapsulated) melatonin (N-acetyl-5-methoxytryptamine) for drug delivery and controlled release on the depletion of inflammatory mediators, as well as oxidative damage in sodium fluoride (NaF)-treated lungs and liver. Hepatic and bronchial damage was induced in Swiss albino rats with a single acute ingestion of NaF (48 mg/kg body weight, oral gavage). NaF exposure caused the generation of reactive oxygen species (ROS); upregulation of TNF-α and TGF-β; decreased activities of antioxidant systems (glutathione, glutathione-S-transferase, superoxide dismutase, catalase), succinate dehydrogenase, membrane microviscosity, and membrane potential; increased activity of lipid peroxidation and nicotinamide adenine dinucleotide hydride oxidase; and increased hepatic and nephrite toxicities (P

Plant growth-promoting rhizobacteria (PGPR) are soil bacteria, colonizing rhizospheric region of plants, which have the ability to enhance plant health and promote plant growth by increasing seed emergence, plant weight, and yields to a wide variety of crops either through direct action or via biological control of plant diseases. PGPR improve plant growth by either fixing atmospheric nitrogen; solubilizing insoluble phosphates and iron and producing plant growth regulators (PGRs) like auxins, gibberellins, cytokinins, etc.; or suppression of deleterious root-colonizing microorganisms including plant pathogens through antibiosis, i.e., production of fungitoxic compounds, competition with pathogenic microorganisms for nutrients by producing siderophores, or niche exclusion. Indiscriminate use of different chemicals in the form of fertilizers and pesticides targeting to increase the agricultural produce for ever-increasing population outburst has led to the contamination of the groundwater, soil, and sediments. Accretion of diversified range of chemicals in significant quantities has a direct impact not only on the living beings but also on the environment. The ecological balance of the soil microorganisms has been distorted which show the negative impact on their rhizospheric competence. When exogenous PGPR are applied in this pesticide-infested soil, they not only hardly show their plant growth-promoting or disease-suppressing activities but also might not survive at all. Isolation of native PGPR from the pesticide-challenged rhizospheric soils mostly shows pesticide-tolerant/degrading properties. These PGPR might show the rhizospheric competence in similar pesticide-infested soil. These strains easily acclimatize in the pesticide-contaminated microenvironment in soil and show their plant growth-promoting and pathogen-suppressive activities.

Wild edible mushrooms are utilized by ethnic people in rural communities of India as food and medicinal purposes. The aim of the present study was to estimate biochemical constituents and antioxidant activities of two wild edible mushrooms viz. Russula gnathangensis and Ramaria thindii isolated from Sikkim Himalaya. Methanolic extract of oven dried mushroom powders were used for the estimation of polyphenol, flavonoid, tannin, lycopene and ­β-carotene contents and determination of antioxidant activities though protein and carbohydrate contents were estimated using aqueous extracts. All the chemical constituents were measured on dry wt. basis. Protein contents of R. thindii and R. gnathangensis were 185.8 μg/mg and 164.7 μg/mg, respectively. R. thindii showed 191.7 μg/mg carbohydrate content whereas R. gnathangensis showed 140.5 μg/mg. Polyphenol and flavonoid contents of R. thindii were 7.9 μg/mg and 1.0 μg/mg, respectively whereas R. gnathangensis showed 4.25 μg/mg polyphenol and 0.75 μg/mg flavonoid contents. Tannin contents of R. thindii and R. gnathangensis were 2.35 and 1.46 μg/mg, respectively. R. thindii showed better lycopene content but both the species showed similar amount of ­β-carotene. Both the species showed high amount of antioxidant activities. These wild mushrooms may fulfill the nutritional requirement of the local ethnic people if properly conserved.

Recent evidences suggest that cerebral ischemia-reperfusion insult plays significant role in pathogenic diseases like Alzheimer's disease (AD) and other neurodegenerative diseases. Toxic reactive oxygen species (ROS) generated by induced oxidative stress in the episodes of cerebral ischemia-reperfusion (CIR) plays major role in neurodegeneration. As the prime source of ROS generation, neuronal mitochondria, the cellular energy metabolic centre experience severe damage because of CIR-induced oxidative stress. The process of mitochondrial dysfunction is accelerated by CIR that may pave the pathway for neurodegeneration in AD among aged individuals. Prevention of CIR injury may be a shunt in order to minimize the risk of dementia of Alzheimer's type in aged individuals. The use of chemical antioxidants in CIR is not suitable as the blood- brain barrier (BBB) doesn't allow the entry of molecules from blood circulation into the brain. Thus L-ascorbic acid loaded polylactide nanocapsules were prepared and fed orally to assess the role of nanocapsulated ascorbic acid (NAA) against CIR induced oxidative injury in mitochondrial region of rat brains. Mitochondrial injury was assessed by the extent of lipid peroxidation and in situ antioxidant enzyme status. The levels of cytochrome c (cyt c), cyclooxygenase- 2 (COX-2) and iNOS were determined. Results showed that in comparison to free ascorbic acid (AA), NAA exerted better protection to the brain mitochondria by preventing oxidative damage in ROS mediated CIR injury.

Triple negative breast cancer (TNBC) is one of the most common invasive malignancies among women, associated with poor prognosis. Standard chemotherapy targets all dividing cells, resulting in dose-limiting toxicities. In this study, we demonstrated a strategy of encapsulating a hydrophobic synthetic compound, nifetepimine, having anticancer properties, in poly (lactic-co-glycolic acid) nanoparticles to increase selectivity of drug to cancerous cells with minimum toxicity towards normal cells. Nanoencapsulated nifetepimine (30-100nm) having loading and encapsulation efficiency of 7.45% and 75% respectively, was successfully internalized inside TNBC cells upon sustained release resulting in apoptosis. An in vivo bio-distribution study indicated that nanonifetepimine selectively accumulated into breast tumor sites of mice, primarily due to prolonged blood circulation time and binding of nifetepimine to epidermal growth factor receptor that remains overexpressed in most of the TNBC tumors. Moreover, we observed significant reduction in breast tumor volume with improved survival implying high tumor targetability of nanonifetepimine.

Background Cyclooxygenase-2 (Cox-2) is frequently overexpressed in cervical carcinoma, but little is known about its altered serum concentration. Hence, this study evaluates clinical utility of cellular and serum level of Cox-2 enzyme in cervical cancer. Methods The expression of Cox-2 was evaluated in cervical tissues and serum samples collected from normal controls (n = 100; n = 68), cervical intraepithelial neoplasia patients (CIN, n = 67; n = 12), and invasive squamous cell carcinoma patients (SCCs; n = 153; n = 127) by immunohistochemical and enzyme-linked immunosorbent assay (ELISA) analyses. Results The significant cytoplasmic overexpression of Cox-2 was noted in 50.7% of CIN and 69.9% of SCCs as compared with normal (P = 0.0001). Serum level of Cox-2 was also found to be elevated both in CIN (median 4.35 ng/ml) and in SCCs (median 19.39 ng/ml) with respect to normal (median 0.44 ng/ml; P = 0.0001), respectively. The ROC analysis revealed the potential of serum Cox-2 over its cellular expression to distinguish CIN and SCCs from normal. Conclusion Augmented Cox-2 activity is implicated in the pathogenesis of cervical cancer, and its serum level could serve a potential to distinguish this malignancy. Therefore, it is suggested that serum Cox-2 may be useful in monitoring the diagnosis and treatment outcome of patients.

Oxidative stress a major cause of fluoride induced toxicity and mitochondrial impairment in common in experimental rats during chronic exposure of fluoride. Attempts have been made in the present experiment to diminish oxidative damage, combined therapy with (+)-catechin hydrate (an antioxidant) and sodium meta borate (chelator) were used. Fluoride intoxication in rats was performed by using 13mg/kg NaF and both antioxidant CH and chelator SMB were used at a concentration of 8.98μM/kg body weight. Mixture of CH and SMB in free or in PLGA nanocapsule encapsulated form were prepared. The efficacies of those formulations were tested in combating free radical mediated oxidative insult produced by sodium fluoride (NaF). The amalgamated therapy used in this experiment was shown to reduce fluoride levels in liver, brain and kidney from 9.5, 5.5, 6.3μg/g to 4.6, 2, 2.6μg/g, respectively. Our result indicated that the combined chelator and antioxidant therapy in nanocapsulated drug delivery system could provide a projection in combating fluoride induced mitochondrial impairment in rat model.

Liquid crystal-gold nanoparticle (GNP-LC) composite were accessed via covalent coupling of 4-mercaptophenol capped gold nanoparticles (GNP) and a carboxylic acid functionalised rod shaped mesomorphic Schiff base (LC). The Schiff base ligand and the composite both showed schlieren texture with distinct difference in phase transition temperature. The X-ray diffraction study suggested a smectic C mesophase for the compounds. The LC and GNP-LC, both exhibited room temperature photoluminescence when excited at 264nm.The transmission electron micrography revealed the composite to be nearly monodisphersed having almost spherical shape of ~ 2nm size. The composite material in addition to being mesomorphic exhibited pronounced AC conductivity in the semiconductor domain and antioxidant activity.

Cerebral ischemia-reperfusion is a classic example of reactive oxygen species (ROS) mediated acute damage to brain. Post-ischemic reperfusion induced oxygen free radicals production causes damage to brain cell mitochondria. Antioxidants like quercetin (Qc) have potentials to manage oxidative stress related pathophysiology. However low oral bioavailability and poor cell membrane permeability restrict its therapeutic efficacy. To overcome these hurdles mitochondria specific delivery of Qc nanocapsules was designed to efficiently counteract cerebral ischemia-reperfusion induced cell death and neurodegeneration in young and aged rats. The orally deliverable quercetin loaded polymeric nanocapsules (N1QC) were made mitochondria specific by using triphenylphosphonium cation as one of the matrix components. N1QC demonstrated higher brain uptake and remarkable mitochondrial localization post cerebral ischemia-reperfusion. This unique controlled mitochondrial delivery of quercetin ameliorated histopathological severity by preserving mitochondrial structural and functional integrity through sequestering ROS thus modulating mitochondrial ROS mediated apoptotic cell death in young and aged rats.

Aims: The aims of our work were to investigate the controlling role and the efficacy of nanocapsulated quercetin drug delivery system on the decrement of inflammatory mediators such as MMP-13 in diethyl nitrosamine (DEN)-induced hepatocarcinogenesis. Materials & methods: Hepatocellular carcinoma was developed in the Swiss albino rats by the exposure of DEN. DEN administration caused the generation of reactive oxygen species, upregulation of TNF-α, IL-6, activation of MMP-13, severe oxidative damage, hyperplastic nodules with preneoplastic lesions and the histopathological changes in rat liver. Results & conclusion: Nanocapsulated quercetin treatment restricted all alterations in DEN-mediated development of hepatocarcinogenesis. Therefore, it may be concluded that nanocapsulated quercetin may be accepted as a potent therapeutic formulation in preventing DEN-mediated hepatocarcinogenesis. Original submitted 4 January 2013; Revised submitted 2 December 2013

Cupric oxide (CuO) nanoparticles have been successfully synthesized via low-temperature thermal decomposition of bis(dimethylglyoximato)copper(II) complex. The synthesized nanoparticles were characterized by powder X-ray diffraction (XRD), transmission electron microscopy (TEM), dynamic light scattering (DLS) and Raman spectroscopy. The powder XRD pattern furnished evidence for a monoclinic structure of CuO with average crystallite size 28.9 nm. The TEM image showed aggregation of some quasi-spherical nanoparticles. The Raman spectrum showed three characteristic modes of CuO which further affirmed the phasic purity of the obtained material. The CuO nanoparticles studied in an in vitro system using the modified diphenylpicrylhydrazyl (DPPH) method exhibited pronounced antioxidant activity.

Hepatocellular carcinoma (HCC) is the fourth leading cause of cancer-related death worldwide, and satisfactory treatment is unavailable. Doxorubicin (DOX) is commonly used for HCC treatment with very limited success and serious side-effects such as cardiotoxicity and non-cancer cell cytotoxicity. Methods In the present investigation, a new nano-sized hydroxyapatite (HAp)-based drug-delivery system was successfully developed with nano-sized hydroxyapatite (HAp) (sizes 5–30 nm) and synthesised with a Ca/P molar ratio of 1.67. After thorough in vitro characterisation, these nano-HAP particles were loaded/intercalated with DOX (50–60% encapsulation efficiency), and thorough characterisation of the size, shape and morphology of the particles was performed. Results The average drug-loaded nanoparticles had a spherical morphology with a size range of 40–60 nm. The in vitro drug-elution kinetics were examined under different pH conditions to account for the actual pH conditions found in the body environment. The kinetics were observed to be ideal for IV therapy to treat HCC. In vivo experiments using animal models demonstrated very promising results in terms of relative liver weight changes and histopathology. Ultimately, ‘high-dose’ HAp-DOX produced maximum suppression of hyperplastic nodules and a minimum number of preneoplastic lesions. In summary, our results indicate that this new formulation is an efficient, safe and reliable treatment method for HCC.

Multiply twinned phase pure copper nanoparticles were successfully synthesized without any inert gas protection via a low-temperature thermal decomposition of a new precursor copper malonate, [CuCH 2 C 2 O 4 ]·2H 2 O in the presence of surfactant stabilizers oleylamine (C 18 H 35 NH 2 ) and triphenylphosphine ((C 6 H 5 ) 3 P). The synthesized nanoparticles were characterized by powder X-ray diffraction (XRD), transmission electron microscopy (TEM), FT-IR and UV–vis spectroscopy. Freshly prepared copper nanoparticles showed surface plasmon resonance (SPR) around 574 nm. TEM studies revealed a five-fold multiply twinned morphology of decahedral shape for the copper nanoparticles. Powder XRD pattern furnished evidence for a face-centered cubic crystal structure of pure metallic copper having an average crystallite size of about 35 nm.

GNP-LC hybrid incorporating giant gold nanoclusters was accessed with tailor made liquid crystalline Schiff base bearing amine function as reactive end group without using any external reducing agent. The liquid crystal phase (smectic C) of the Schiff base remained unaltered in the hybrid material. The ligand and hybrid both exhibited interesting concentration-dependent fluorescence quenching of 9,10-diphenylanthracene when excited at 345 nm wavelength in dichloromethane solution. The GNP-LC hybrid exhibited reasonably good electrical conductivity showing an Arrhenius-type behaviour in the smectic mesophase and a tunnelling behaviour in the crystalline state.

A low-temperature synthesis of tricobalt tetraoxide, Co 3 O 4 nanoparticles was accomplished via thermal decomposition of a new precursor cobalt malonate, [CoCH 2 C 2 O 4 ]· 2H 2 O in presence of surfactant stabilizers oleylamine (C 18 H 35 NH 2 ) and triphenylphosphine ((C 6 H 5 ) 3 P). The synthesized nanoparticles were characterized by powder X-ray diffraction (XRD), transmission electron microscopy (TEM), FT-IR and Raman spectroscopy. Powder XRD pattern furnished evidence for a face-centered cubic structure of Co 3 O 4 with average crystallite size of about 10 nm. TEM studies revealed Co 3 O 4 nanoparticles to be monodispersed, mostly cubic having sizes 4–12 nm. Electron diffraction pattern indicates polycrystalline nature of the synthesized material. Raman spectrum showed four characteristic peaks of Co 3 O 4 which further affirmed the phasic purity of the material.

Tricobalt tetraoxide nanoparticles have been successfully synthesized following a ‘bottom-up’ approach by surfactant-free thermal decomposition of cobalt(II)-tartrate complex obtained by a modified sol–gel route. The synthesized complex was characterized by Fourier transform infrared (FT-IR) spectroscopy, elemental and thermogravimetric-differential thermal analysis (TG–DTA). The nanoparticles were characterized by powder X-ray diffraction (XRD), transmission electron microscopy (TEM), and Raman studies. The powder XRD pattern furnished evidence for a face-centered cubic structure of Co3O4. With the rise in calcination temperature from 400 through 500 to 600 °C, the average crystallite sizes of Co3O4 were found to increase from 28 through 36 to 46 nm. The TEM image revealed a faceted morphology of the as-synthesized Co3O4 nanoparticles. The high-resolution TEM image indicated the interplanar separation to be 0.28 nm which corresponds to the (220) plane in face-centered cubic Co3O4. The electron diffraction (ED) pattern showed single-crystalline nature of the synthesized nanoparticles. Raman spectrum showed four characteristic peaks of Co3O4 which further confirmed the phasic purity of the material.

Zinc oxide nanocrystals were synthesized via thermal decomposition of a new precursor zinc malonate, [ZnCH2C2O4]·2H2O in the presence of surfactant stabilizers oleylamine (C18H35NH2) and triphenylphosphine ((C6H5)3P). The nanocrystals were characterized by X-ray diffraction, transmission electron microscopy, FT-IR and UV–visible spectroscopy. XRD pattern showed a hexagonal wurtzite structure of ZnO having average crystallite size of about 22 nm. The TEM image showed the nanocrystallites constituting the flower-like pattern to be of 25–55 nm sizes. The band gap luminescence was observed at 354 nm (∼3.50 eV) alongwith defect related blue emission at 450 nm (∼2.75 eV).

Gastric ulcer is a multifaceted process that involves reactive oxygen species (ROS) generation, extracellular matrix degradation and mitochondrial damage. Mitochondria play a crucial role for homeostasis of ROS and cell survival. In our study, we investigated the efficacy and mechanism of polymeric nanocapsuled quercetin (NQC) over the free quercetin (QC) molecule in prevention of ethanol-induced gastric ulcer in rat. NQC possessed significantly higher efficacy (~20 fold) than free QC while preventing gastric ulcers. Our data show that prior administration of NQC and/or QC significantly blocked synthesis and secretion of matrix metalloproteinase (MMP)-9 as well as infiltration of inflammatory cells and oxidative damage in rat gastric tissues. As compared to free QC, NQC protected much better the mitochondrial integrity and size along with mitochondrial functions by controlling succinate dehydrogenase and NADH oxidase in rat gastric tissues. In addition, both free QC and NQC down regulated PARP-1 as well as apoptosis during protection against ethanol-induced gastric ulcer. Herein, the effect of NQC was greater than QC on expression of enzymes like cyclooxygenase and nitric oxidase synthase (NOS)-2. We conclude that NQC with greater bioavailability offers significantly higher potency in downregulating MMP-9 and NOS-2 as well as oxidative stress in blocking ethanol-induced gastric ulcer.

Chronic arsenic exposure causes oxidative stress and mitochondrial dysfunction in the liver and brain. The ideal treatment would be to chelate arsenic and prevent oxidative stress. meso-2,3-Dimercaptosuccinic acid (DMSA) is used to chelate arsenic but its hydrophilicity makes it membrane-impermeative. Conversely, quercetin (QC) is a good antioxidant with limited clinical application because of its hydrophobic nature and limited bioavailability, and it is not possible to solubilize these two compounds in a single nontoxic solvent. Nanocapsules have emerged as a potent drug delivery system and make it feasible to incorporate both hydrophilic and lipophilic compounds. Nanoencapsulated formulations with QC and DMSA either alone or coencapsulated in polylactide-co-glycolide [N(QC + DMSA)] were synthesized to explore their therapeutic application in a rat model of chronic arsenic toxicity. These treatments were compared to administration of quercetin or DMSA alone using conventional delivery methods. Both nanoencapsulated quercetin and nanoencapsulated DMSA were more effective at decreasing oxidative injury in liver or brain compared to conventional delivery methods, but coencapsulation of quercetin and DMSA into nanoparticles had a marked synergistic effect, decreasing liver and brain arsenic levels from 9.5 and 4.8 μg/g to 2.2 and 1.5 μg/g, respectively. Likewise, administration of coencapsulated quercetin and DMSA virtually normalized changes in mitochondrial function, formation of reactive oxygen species, and liver injury. We conclude that coencapsulation of quercetin and DMSA may provide a more effective therapeutic strategy in the management of arsenic toxicity and also presents a novel way of combining hydrophilic and hydrophobic drugs into a single delivery system.

Arsenic is a naturally occurring toxicant that causes acute and chronic adverse health effects, including cancer.The study was performed to evaluate the therapeutic efficacy of liposome entrapped flavonoidal quercetin in combating arsenic toxicity mediated oxidative damage in hepatocytes and brain cells in rat model.Hepatic and neuronal cell damage in rats was made by daily arsenic (6 mg/kg b wt, 9 mg/kg b wt and 12 mg/kg b wt) treatment via oral route for four consecutive months. Liposomal quercetin (2.71 mg QC/kg b. wt) were injected s.c. on rats treated with 12 mg/kg b. wt. NaAsO(2) twice a week for four months.Inorganic arsenic deposition was found to be most significant in hepatic (9.32 ± 0.100 µg/g tissue) and neuronal (6.21 ± 0.090 µg/g tissue) cells of rats treated with 12 mg/kg b wt of arsenite. Antioxidant levels in hepatic and neuronal cells were reduced significantly by the induction of arsenic. Liposomal quercetin was found most potent for a complete prevention of arsenite-induced reduction in antioxidant levels in the liver and brain of rats. Arsenic induced a substantial increase in hepatic hydroxyproline (HP) and Liposomal quercetin treatment resulted in complete replenishment of the HP level to normal. Liposomal quercetin completely prevented the arsenite-induced upregulation of cytochrome c expression in liver and brain significantly suggesting that the protective effect of Liposomal quercetin could be related to the reduction of arsenic deposition in both the organs.Thus, Liposomal quercetin might prove to be of therapeutic potential against arsenite-induced hepatic and neuronal cell damage in rats.

A new laccase-conjugated antibody is developed, containing laccase as an enzyme marker, obtained from culture supernatant of the basidiomycetous fungi Pleurotus ostreatus. The efficacy of the laccase-conjugated antibody was demonstrated in indirect and direct enzyme immunoassay after using periodate and glutaraldehyde conjugation methods. The assay based on laccase-conjugated antibody is potent to detect antigens when compared with peroxidase conjugated antibody.

Arsenic, the environmental toxicant causes oxidative damage to liver and produces hepatic fibrosis. The theme of our study was to evaluate the therapeutic efficacy of liposomal and nanocapsulated herbal polyphenolic antioxidant quercetin (QC) in combating arsenic induced hepatic oxidative stress, fibrosis associated upregulation of its gene expression and plasma TGF beta (transforming growth factor beta) in rat model. A single dose of arsenic (sodium arsenite-NaAsO(2), 13 mg/kgb.wt) in oral route causes the generation of reactive oxygen species (ROS), arsenic accumulation in liver, hepatotoxicity and decrease in hepatic plasma membrane microviscosity and antioxidant enzyme levels in liver. Arsenic causes fibrosis associated elevation of its gene expression in liver, plasma TGF ss (from normal value 75.2+/-8.67 ng/ml to 196.2+/-12.07 ng/ml) and release of cytochrome c in cytoplasm. Among the two vesicular delivery systems formulated with QC, polylactide nanocapsules showed a promising result compared to liposomal delivery system in controlling arsenic induced alteration of those parameters. A single dose of 0.5 ml of nanocapsulated QC suspension (QC 2.71 mg/kg b.wt) when injected to rats 1h after arsenic administration orally protects liver from arsenic induced deterioration of antioxidant levels as well as oxidative stress associated gene expression of liver. Histopathological examination also confirmed the pathological improvement in liver. Nanocapsulated plant origin flavonoidal compound may be a potent formulation in combating arsenic induced upregulation of gene expression of liver fibrosis through a complete protection against oxidative attack in hepatic cells of rat liver.

Flavonoid is a polyphenolic antioxidant used to treat free radical mediated tissue damage. So flavonoids are the good targets for treatment of many diseases like cancer, toxicity health hazards, neurodegenaration etc. Drug delivery of flavonoid is the novel approach where dosing and drug toxicity can be minimized. Antioxidant Catechin Hydrate (CH) is choosen to make drug delivery vehicle: nanocapsules either alone or in combination with chelator sodium meta borate (SMB) within biodegradable poly-lactide-co-glycolide (PLGA). Sizes of the prepared particles were determined through scanning and atomic force microscopy where less than 50 nm particle sizes were revealed. Bioavailability in serum study determines that shelf life of CH can be increased with nanocapsule delivery compared to free drug. Results demonstrate the efficacy of Nanocapsulated CH to be a potent vehicle for tissue targeting.

Oyster mushroom, Pleurotus ostreatus (Jacq.:Fr.) Kumm. ITCC 3308 (collected from Indian Type Culture Collection, IARI, New Delhi, India, 110012) was grown on dry weed plants, Leonotis sp, Sida acuta, Parthenium argentatum, Ageratum conyzoides, Cassia sophera, Tephrosia purpurea and Lantana camara. Leonotis sp. was the best substrate in fruit body production of P. ostreatus when it was mixed with rice straw (1:1, wet wt/wet wt) for mushroom cultivation. The fruiting time for P. ostreatus was also less on Leonotis sp. than on any other weed substrates tested in the present investigation. T. purpurea was the least suited weed for oyster mushroom cultivation. The main problem of oyster mushroom cultivation on weed substrates was found to be low yield in the second flush that could be overcome by blending weed plants with rice straw. The protein contents of the fruit bodies obtained from Cassia sophera, Parthenium argentatum and Leonotis sp. were not only better than rice straw but also from the rice straw supplemented weeds.

Pleurotus ostreatus (Florida), ITCC 3308 produces approximately 9.0 U/ml extracellular cellobiose dehydrogenase (CDH) in cellulose medium after 7 days of growth. However, no activity could be detected if the assay was done with cellobiose as the substrate and 2,6-dichlorophenol indophenol (DPIP) as the electron acceptor in absence of any laccase inhibitor. Kinetic study showed that V(max)/K(m) value was very high for rDPIP (reduced 2,6-dichlrophenol indophenol) oxidation by laccase. Oxygen consumption rate of rDPIP oxidation by the enzyme was found to be highest among all the tested substrates. The present study indicated that rDPIP was a good substrate for laccase. Therefore, caution is needed to measure CDH activity by monitoring DPIP reduction in a system where laccase is likely to be present.

Measurements of the specific activities of the representative enzymes of different pathways linked to carbohydrate metabolism indicate that glycolysis and TCA cycles are the major route of carbohydrate catabolism in the sporulating phase of fruiting body development in Pleurotus ostreatus. Enzymes of the pentose phosphate pathway always showed lower specific activities as compared to those of the enzymes of the glycolytic pathway. The activity of NADP linked glutamate dehydrogenase which is known to be an anabolic enzyme decreased drastically in sporulating fruiting bodies and in spore containing gill tissue (spore bearing structure). Mannitol dehydrogenase activity declined significantly in the sporulating phase of P. ostreatus. The high rate of metabolism during sporulation was further supported by a lower rate of gluconeogenesis at this stage. Concentrations of all the major sugars of the fruiting body (mannitol, glucose and trehalose) decreased in the mature fruiting body and gill tissue. This indicated high catabolic activities at this stage of development.

Bacopasaponin C, an indigenous glycoside, was isolated from Indian medicinal plant Bacopa monniera (b. brahmi) and was tested for antileishmanial properties both in free and in various delivery modes, e.g., niosomes, microspheres, and nanoparticles that are used now as alternatives to more commonly used liposomes. The different vesicles were prepared by published protocols. The percent intercalation of Bacopasaponin C in liposomes, niosomes, and micropspheres determined at its absorption maximal (lambda(max) = 238 nm, epsilon = 8.6 x 10(3) M(-1) x cm(-1)) was found to be 30; for nanoparticles it was 50. At equivalent dose of 1.75 mg/kg body weight, every third day for a total of 6 doses in 15 days, Bacopasaponin C in all the vesicular forms was found to be very active. An inverse linear relationship between the efficacy and the size of the vesicles was established. As analyzed from tissue histology, blood pathology, and specific tests related to normal liver and kidney functions, Bacopasaponin C in each of the four vesicular forms was found to be without any side effects. Thus, because of its indigenous origin and non-toxic nature, Bacopasaponin C could very well be considered for application in the clinic through these alternative delivery modes.

Pleurotus fossulatus (Cooke) Sace is member of oyster mushroom can produced extracellular laccase (benzenediol: oxygen oxidoreductase; EC 1.10.3.2) in submerged fermentation. To analyze the optimum production for laccase P. fossulatus was cultured both in stationary and shaking condition in different media. Partial purification of laccase was done after 0-80% ammonium sulphate precipitation, followed by DEAE (Diethylaminoethyl) Sephadex (A-50) anion exchange chromatography. Potato-sucrose peptone (PSP) medium and Potato-dextrose (PD) medium showed highest laccase production in shaking and stationary conditions, respectively. Though the time required for optimum laccase production in stationary condition was much more than the shaking condition but the amount of laccase was about 2.75t greater in former condition. The laccase produced in stationary condition was more stable than the enzyme produced in shaking condition. The partially purified enzyme showed highest affinity towards o-dianisidine than guaiacol and ABTS (2,2'-Azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) as evidenced by their K(m). The physico-chemical properties of the laccase suggested the significance of this enzyme in industrial applications.

The study investigates the potential of substitution of the conventional carbohydrate nutrient (cellulose) in media with cheap agro-residues for cellobiose dehydrogenase production by Termitomyces clypeatus (CDHtc) under submerged conditions. Different agro-residues tested for enzyme production were characterized using FTIR and XRD analysis. As CDHtc production was highest with tamarind kernel powder (TKP), it was selected for process optimizations through shake-flask fermentations. The optimized parameters were then applied to batch cultures in a 5 L bioreactor that gave enzyme yield (57.4 U mL⁻¹) similar to that obtained under shake-flask fermentations (57.05 U mL⁻¹). The study also made an attempt to predict CDHtc production with respect to time of fermentation and mycelial growth. The specific growth rate and carrying capacity of the mycelia were also determined, and the values lie in the ranges of 0.024-0.027 h⁻¹ and 7.2-7.1 mg mL⁻¹, respectively.

During sporulation, laccase activity of Pleurotus florida decreased to a minimum level in spite of increase in the number of isozymes. An endogenous laccase substrate was detected especially in the gill structure of the sporophore, which competitively inhibited oxidation of guaiacol by the enzyme during in vitro assay. Appearance of the laccase substrate in the gill structure may be linked with the sporulation phenomenon.

Disturbance of calcium homeostasis and unregulated increase in intracellular calcium have been implicated in cell damage and cell death in the central nervous system in particular. To determine the specific pathway(s) of cerebral Ca transport of importance in a pathophysiological situation, we have measured long term Ca flux in brain in vivo in rat, and developed a kinetic model incorporating physiologically relevant pathways of cerebral Ca transport. 45Ca was injected into a tail vein in conscious rats. Plasma 45Ca was monitored up to 4 days post-injection and 45Ca uptake determined in samples of cerebrospinal fluid (CSF) and at several sites of brain at euthanasia. Uptake of the tracer by tissues peaked after 1 h, isotopic equilibration taking longer. The uptake at hippocampus was the highest. Computer simulation of the kinetics of the plasma, CSF and tissue data was performed using a compartmental model, which included two subcompartments (intra- and extra-cellular) and two pathways from plasma to the brain: directly across the blood-brain barrier and via CSF, which included a delay. The analysis based on this model enabled estimation of the fractional rates of transport of Ca to cerebral and noncerebral tissues across all the barriers of the model.

The uptake of 45 Ca 2+ by different regions of the brain was determined in anaesthetised rat after infusing angiotensin II (AII; 14 μ g in 1 ml) through the carotid artery. The control animals (C) were infused with physiological saline. 45 Ca 2+ was injected in the tail vein 2 min after the infusion of AII or saline. Plasma radioactivity was monitored until euthanasia at different times post injection when the cerebral uptake of 45 Ca 2+ was determined. While the plasma turnover of 45 Ca 2+ was similar in AII and C animals, the uptake was markedly higher in AII vs. C at the different brain sites at all times. Kinetic analysis suggested that AII induced a short term increase in Ca 2+ transport across the blood–brain barrier and also in the cellular uptake. The effect of AII was receptor mediated being abolished by saralasin, an AII antagonist, and appeared to be at least in part independent of its pressor effect.

Aim: Hepatocellular carcinoma (HCC) has no successful pharmacotherapeutic remedy. The aim of this study was to ascertain whether ginkgolide B (GB)-loaded polymeric nanocapsules can prevent diethylnitrosamine (DEN)-induced HCC in rats. Materials & methods: GB was fabricated in two types of nanocapsules of which one was polyethylene glycol coated (N1GB) and the other was uncoated (N2GB). These nanocapsules were orally gavaged during DEN-induced HCC development in rats. Results: Nanocapsulation of GB enabled aqueous suspension and slow time-dependent release of the compound. Anticarcinogenic potential of N2GB was reflected by its ability in the management of DEN-induced reactive oxygen species generation, mitochondrial dysfunction, p53, NF-κB, inducible nitric oxide synthase, COX-2 and VEGF expressions, and induction of apoptosis in cancer cells in the rat liver. Conclusion: Positive zeta-potential on N2GB surface might have offered higher hepatic accumulation of GB, especially at the electron-dense organelle mitochondria. Mitochondria protection against DEN-induced oxidative damage ensured HCC prevention. Original submitted 27 June 2012; Revised submitted 4 December 2012; Published online 7 June 2013

Oxidative stress, due to the generation of reactive oxygen species, is a major factor in cerebral ischemic damage and changes the activities of antioxidant enzymes and substantially influences the aging process. Free chemical antioxidant is almost ineffective to treat brain ischemia as blood-brain barrier exists in between blood and brain interstitial fluid, limiting component to pass from the circulation into cerebral region. Different compounds have been tested in vivo in different vesiculated forms to prevent cerebral ischemia. Nanoparticle-encapsulated drug treatment resulted in a significant protection of the antioxidant enzymes in both young and old rats. Nanocapsulated drug treatment causes a substantial protection against cerebral ischemia-reperfusion-induced oxidative damage to all parts of brain specifically hippocampal regions of all age groups of rat brain.

Oxidative stress, due to the generation of reactive oxygen species, is a major factor in cerebral ischemic damage and changes the activities of antioxidant enzymes and substantially influences the aging process. Free chemical antioxidant is almost ineffective to treat brain ischemia as blood-brain barrier exists in between blood and brain interstitial fluid, limiting component to pass from the circulation into cerebral region. Different compounds have been tested in vivo in different vesiculated forms to prevent cerebral ischemia. Nanoparticle-encapsulated drug treatment resulted in a significant protection of the antioxidant enzymes in both young and old rats. Nanocapsulated drug treatment causes a substantial protection against cerebral ischemia-reperfusion-induced oxidative damage to all parts of brain specifically hippocampal regions of all age groups of rat brain.