EVALUATION OF DIFFERENT CULTURE MEDIA FOR ENHANCED PRODUCTION OF PSEUDOMONAS AERUGINOSA (MTCC NO 2453) BIOMASS AND ITS PROTEINS

Objective: Microorganisms, especially bacteria and its proteins have proven to be potential anti-cancer agents as they selectively attack the tumor cells or tumor micro-environments. The extract of Pseudomonas aeruginosa found to contain proteins that have shown promising anticancer activity. In this work, it was attempted to increase the biomass and trigger the total protein fraction of Pseudomonas aeruginosa (MTCC 2453).Methods: The organism was cultivated in three different such as Luria-Bertani (LB) broth, minimal medium9 (M9), super broth medium (SB) and asparagine-proline (AP) broth. Asparagine proline broth was selected as it has shown high cell growth rate. The media was further optimized by the addition of NaHCO3 and copper sulphate to trigger the protein production. Optimized Aspergine proline broth has achieved highest cell biomass. After the shake flask culture, the overnight grown culture in optimized AP medium was further grown in a 5 L bioreactor by fed-batch cultivation to achieve higher cell densities.Results: The highest protein production was achieved at 40 ° C. Highest biomass and protein content was observed at pH 8 while lowest biomass was produced at pH 2. A gradual increase in biomass content observed from 12 h towards to 48 h.Conclusion: High biomass and proteins content and of Pseudomonas aeruginosa (MTCC 2453) can be produced in optimized asparagine-proline broth. Further the extract is purified to produce novel anti-cancer proteins

Solid-State Nuclear Magnetic Resonance of Copper-Amyloid Beta, Amylospheroids, Fast Magic Angle Spinning

Molecular level structural examination of Cu2+ binding to 40-residue Alzheimer’s amyloid β (Aβ(1-40)) peptides and the resultant production of reactive oxygen species (ROS) was studied by SSNMR and other techniques. Aβ(1-40) peptide is one of the primary component of senile plaques in Alzheimer’s disease (AD) and was shown that Cu ions (400 µM) are accumulated in plaque deposits and bind Aβ peptides. The Cu-Aβ complex is believed to trigger the production of ROS causing oxidative stress. Despite oxidative stress being considered one of the probable mechanisms of AD, detailed binding structure of Cu2+-Aβ fibrils and the molecular level mechanism of the production of ROS in presence of cellular reductants are still largely unknown. In our work, we try to understand the molecular level details of Cu2+ binding to Aβ(1-40) fibrils, the mechanism of ROS production and subsequent binding of Cu ions with Aβ(1-40) fibrils. Initial signal quenching studies by 13C SSNMR and molecular dynamics simulations showed Cu2+ binding to Aβ(1-40) fibrils at Nε in His-13, His-14 and carboxyl groups in Val-40 as well as Glu sidechains (Glu-3,Glu-11, and/or Glu-22). In the presence of physiological amounts of biological reductant ascorbate Cu2+-Aβ(1-40) complex generates ~60 μM H2O2. During the production of H2O2, Cu2+-Aβ(1-40) fibrils undergoes cyclic redox reaction (Cu2+ ↔ Cu+ )- Aβ(1-40) without any alteration to the fibrils. SSNMR experiments revealed that Cu+ ions are bound to Nδ in His-13 and His-14 during the ROS production. In the second part, the site-specific molecular level structural features of the synthetic Aβ(1-42) amylo-spheroids (ASPD) which is a structural/morphological analog of native ASPD extracted from AD patients is discussed. ASPD represents a class of highly cytotoxic and high mass (>100 kDa) spherical aggregates which have distinct surface tertiary structure and induce degeneration of mature neurons through a different mechanism from other non-fibrillar Aβ assemblies and considered to play a primary role in AD. In the last part, sensitivity and resolution enhancement in 13C and 1H SSNMR is demonstrated by paramagnetic relaxation assisted condensed data collection method, ultra-fast MAS (≥ 78kHz) and selective deuteration using stereo array isotope labeling (SAIL) in a high magnetic field

<i style="">Portulaca oleracea</i> L. extract ameliorates the cisplatin-induced toxicity in chick embryonic liver

185-189 Cisplatin, a cytotoxic agent used in treating cancer, at high doses induces hepatotoxicity. In this study, we investigated the protective role of aqueous extract of aerial parts of Portulaca oleracea L. (Po) against cisplatin-induced hepatotoxicity in chick embryonic liver. A group of 12 day old chick embryos, acclimatized to laboratory conditions were treated with a single dose of cisplatin (100 µg), while another group received Po extract at different doses (1 and 3 mg) 6 h prior to cisplatin treatment. The biochemical parameters were estimated after 24 and 72 h of incubation. A dose-dependent increase in biochemical parameters, such as alanine transaminase, aspartate transaminase, alkaline phosphatase, lactate dehydrogenase, malondialdehyde levels and a decrease in antioxidant enzymes levels like superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase, glutathione-s-transferase and reduced glutathione were observed in cisplatin-treated animals, indicating a definite damage to the liver tissue. Pre-treatment with Po extract was found to provide significant protection against cisplatin-induced hepatotoxicity, as evident by the recovered levels of the altered changes in the measured biochemical parameters. </smarttagtype

Embryo protective effect of pomegranate (Punica granatum L.) fruit extract in adriamycin-induced oxidative stress

106-111The possible protective role of pomegranate (Punica granatum L.) fruit extract which has shown antioxidant capacity higher than that of red wine and green tea was evaluated against adriamycin-induced oxidative stress in chick embryos. Adriamycin (ADR), an anthracycline broad spectrum of chemotherapeutic drug is used for the treatment of variety of cancers; however, its prolonged use is limited by an irreversible, dose-dependant and progressive cardiomyopathy, hepatotoxicity and general toxicity to other organs in human beings, due to oxidative stress. The morphological changes (malformation of different organs), changes in body weight, volume of amniotic fluid (AF) and biochemical parameters of AF were studied after 24 and 48 h of incubation by comparing ADR alone and pomegranate fruit extract pretreated groups with their respective controls of 12 days old chick embryos. ADR alone at a dose of 70 μg/egg showed a significant dose versus time- dependent reduction in body weight, volume of AF. A dose-related increase in embryo gross morphological deformities and significant changes in the levels of biochemical parameters in AF were observed in ADR-treated group. These changes were significantly ameliorated to normal by pre-administration of pomegranate fruit extract at a dose of 200 µg/egg. Thus, the present study demonstrated the embryo protective nature of pomegranate fruit extract against ADR-induced oxidative stress

Metal-Free Oxidative C–C Coupling of Arylamines Using a Quinone-Based Organic Oxidant

A variety of arylamines are shown to undergo oxidative C–C bond formation using quinone-based chloranil/H⁺ reagent as the recyclable organic (metal-free) oxidant system to afford benzidines/naphthidines. Arylamines (3°/2°) designed with various substituents were employed to understand the steric as well as electronic preferences of oxidative dimerization, and a mechanism involving amine radical cation has been proposed. The tetraphenylbenzidine derivative obtained via oxidative C–C coupling has been further converted to blue-emissive hole-transporting material via a simple chemical transformation. This study highlights the preparation of novel HTMs in a simple, economic, and efficient manner

Metal-Free Oxidative C–C Coupling of Arylamines Using a Quinone-Based Organic Oxidant

A variety of arylamines are shown to undergo oxidative C–C bond formation using quinone-based chloranil/H⁺ reagent as the recyclable organic (metal-free) oxidant system to afford benzidines/naphthidines. Arylamines (3°/2°) designed with various substituents were employed to understand the steric as well as electronic preferences of oxidative dimerization, and a mechanism involving amine radical cation has been proposed. The tetraphenylbenzidine derivative obtained via oxidative C–C coupling has been further converted to blue-emissive hole-transporting material via a simple chemical transformation. This study highlights the preparation of novel HTMs in a simple, economic, and efficient manner

Structural Insight into an Alzheimer’s Brain-Derived Spherical Assembly of Amyloid β by Solid-State NMR

Accumulating evidence suggests that various neuro­degenerative diseases, including Alzheimer’s disease (AD), are linked to cytotoxic diffusible aggregates of amyloid proteins, which are metastable intermediate species in protein misfolding. This study presents the first site-specific structural study on an intermediate called amylo­spheroid (ASPD), an AD-derived neurotoxin composed of oligomeric amyloid-β (Aβ). Electron microscopy and immunological analyses using ASPD-specific “conformational” antibodies established synthetic ASPD for the 42-residue Aβ(1–42) as an excellent structural/morphological analogue of native ASPD extracted from AD patients, the level of which correlates with the severity of AD. ¹³C solid-state NMR analyses of approximately 20 residues and interstrand distances demonstrated that the synthetic ASPD is made of a homogeneous single conformer containing parallel β-sheets. These results provide profound insight into the native ASPD, indicating that Aβ is likely to self-assemble into the toxic intermediate with β-sheet structures in AD brains. This approach can be applied to various intermediates relevant to amyloid diseases