Recycling of the Posttermination Complexes of Mycobacterium smegmatis and Escherichia coli Ribosomes Using Heterologous Factors

In eubacteria, ribosome recycling factor (RRF) and elongation factor G (EFG) function together to dissociate posttermination ribosomal complexes. Earlier studies, using heterologous factors from Mycobacterium tuberculosis in Escherichia coli revealed that specific interactions between RRF and EFG are crucial for their function in ribosome recycling. Here, we used translation factors from E.coli,Mycobacterium smegmatis and M. tuberculosis, and polysomes from E. coli and M. smegmatis, and employed in vivo and in vitro experiments to further understand the role of EFG in ribosome recycling. We show thatE. coli EFG (EcoEFG) recycles E. coli ribosomes with E. coli REF (EcoRRF), but not with mycobacterial RRFs. Also, EcoEFG fails to recycle M. smegmatis ribosomes with either EcoRRF or mycobacterial RRFs. On the other hand, mycobacterial EFGs recycle both E. coli and M. smegmatis ribosomes with either of the RRFs. These observations suggest that EFG establishes distinct interactions with REF and the ribosome to carry out ribosome recycling. Furthermore, the EFG chimeras generated by swapping domains betweenmycobacterial EFGs and EcoEFG suggest that while the residues needed to specify the EFG interaction with REF arelocated in domains IV and V. those required to specify its interaction with the ribosome are located throughout the molecule. (C) 2010 Elsevier Ltd. All rights reserved.

Spatial Analysis and Characterisation of Lentic Ecosystems: A Case Study of Varthur Lake, Bangalore

Lentic ecosystems vital functions such as recycling of nutrients, purification of water, recharge of groundwater,augmenting and maintenance of stream flow and habitat provision for a wide variety of flora and fauna along with their recreation values necessitates their sustainable management through appropriate conservation mechanisms. Failure to restore these ecosystems will result in extinction of species or ecosystem types and cause permanent ecological damage. In Bangalore, lentic ecosystems (for example lakes) have played a prominent role serving the needs of agriculture and drinking water. But the burgeoning population accompanied by unplanned developmental activities has led to the drastic reduction in their numbers (from 262 in 1976 to 81). The existing water bodies are contaminated by residential, agricultural, commercial and industrial wastes/effluents. In order to restore the ecosystem, assessment of the level of contamination is crucial. This paper focuses on characterisation and restoration aspects of Varthur lake based on hydrological, morphometric, physical-chemical and socio-economic investigations for a period of six months covering post monsoon seasons. The results of the water quality analysis show that the lake is eutrophic with high concentrations of phosphorous and organic matter. The morphometric analysis indicates that the lake is shallow in relation to its surface area. Socio-economic analyses show dependence of local residents for irrigation, fodder, etc. These analyses highlight the need and urgency to restore the physical, chemical and biological integrity through viable restoration and sustainable watershed management strategies, which include pollution abatement, catchment treatment, desilting of the lake and educating all stakeholders on the conservation and restoration of lake ecosystems.

Phyllosphere of Cotton as a Habitat for Diazotrophic Microorganisms

Positive nitrogenase activities ranging from 0.18 to 0.78 nmol of C2H4 cm−2 h−1 were detected on the leaf surfaces of different varieties of cotton (Gossypium hirsutum L. and G. herbaceum L.) plants. Beijerinckia sp. was observed to be the predominant nitrogen-fixing microorganism in the phyllosphere of these varieties. A higher level of phyllosphere nitrogen-fixing activity was recorded in the variety Varalaxmi despite a low C/N ratio in the leaf leachates. Leaf surfaces of the above variety possessed the largest number of hairy outgrowths (trichomes) which entrapped a majority of microbes. Immersion of plant roots in nutrient medium containing 32Pi led to the accumulation of label in the trichome-borne microorganisms, thereby indicating a possible transfer of nutrients from leaf to microbes via trichomes. Extrapolation of acetylene reduction values suggested that 1.6 to 3.2 kg of N ha−1 might be contributed by diazotrophs in the phyllosphere of the variety Varalaxmi during the entire growth period.

Some experimental observations on the use of activated sludge as fertilizer for fish culture

Experiments were conducted in cement cisterns to find out the effect of adding different dosages of activated sludge on fish growth and plankton production. Three dosages of sludge,viz., 62·5 gm., 125 gm. and 250 gm. per 240 litres of water were used. Fingerlings ofCyprinus carpio, Cirrhina mrigala andCatla catla responded positively,C. mrigala showing maximum growth. The results indicate that the sludge has a direct influence on increasing growth of fish and production of plankton due to the release of nutrients into the water. The increase in plankton content stops after about 30 days. When greater quantities of sludge were added in the cisterns, fish mortality took place.

Microstructure and damping behaviour of consolidated magnesium chips

Chips produced by turning a commercial grade pure magnesium billet were consolidated by solid state recycling technique of cold compaction followed by hot extrusion. The cold compacted billets were extruded at four different temperatures: 250 degrees C, 300 degrees C, 350 degrees C and 400 degrees C. For the purpose of comparison, cast magnesium (pure) billets were extruded under similar conditions. Extruded products were characterized for damping properties. Damping capacity and dynamic modulus was measured as a function of time and temperature at a fixed frequency of 5 Hz 10 to 14% increase in damping capacity was observed in chip consolidated products compared to reference material. Microstructural changes after the temperature sweep tests were examined. Chip boundaries present in consolidated products were observed to suppress grain coarsening which otherwise was significant in reference material. The present work is significant from the viewpoint of recycling of machined chips and development of sustainable manufacturing processes. (C) 2012 Elsevier B.V. All rights reserved.

An ultrastable conjugate of silver nanoparticles and protein formed through weak interactions

In recent years, silver nanoparticles (AgNPs) have attracted significant attention owing to their unique physicochemical, optical, conductive and antimicrobial properties. One of the properties of AgNPs which is crucial for all applications is their stability. In the present study we unravel a mechanism through which silver nanoparticles are rendered ultrastable in an aqueous solution in complex with the protein ubiquitin (Ubq). This involves a dynamic and reversible association and dissociation of ubiquitin from the surface of AgNP. The exchange occurs at a rate much greater than 25 s(-1) implying a residence time of <40 ms for the protein. The AgNP-Ubq complex remains stable for months due to steric stabilization over a wide pH range compared to unconjugated AgNPs. NMR studies reveal that the protein molecules bind reversibly to AgNP with an approximate dissociation constant of 55 mu M and undergo fast exchange. At pH > 4 the positively charged surface of the protein comes in contact with the citrate capped AgNP surface. Further, NMR relaxation-based experiments suggest that in addition to the dynamic exchange, a conformational rearrangement of the protein takes place upon binding to AgNP. The ultrastability of the AgNP-Ubq complex was found to be useful for its anti-microbial activity, which allowed the recycling of this complex multiple times without the loss of stability. Altogether, the study provides new insights into the mechanism of protein-silver nanoparticle interactions and opens up new avenues for its application in a wide range of systems.

Three-dimensional plotted hydroxyapatite scaffolds with predefined architecture: comparison of stabilization by alginate cross-linking versus sintering

Scaffolds for bone tissue engineering are essentially characterized by porous three-dimensional structures with interconnected pores to facilitate the exchange of nutrients and removal of waste products from cells, thereby promoting cell proliferation in such engineered scaffolds. Although hydroxyapatite is widely being considered for bone tissue engineering applications due to its occurrence in the natural extracellular matrix of this tissue, limited reports are available on additive manufacturing of hydroxyapatite-based materials. In this perspective, hydroxyapatite-based three-dimensional porous scaffolds with two different binders (maltodextrin and sodium alginate) were fabricated using the extrusion method of three-dimensional plotting and the results were compared in reference to the structural properties of scaffolds processed via chemical stabilization and sintering routes, respectively. With the optimal processing conditions regarding to pH and viscosity of binder-loaded hydroxyapatite pastes, scaffolds with parallelepiped porous architecture having up to 74% porosity were fabricated. Interestingly, sintering of the as-plotted hydroxyapatite-sodium alginate (cross-linked with CaCl2 solution) scaffolds led to the formation of chlorapatite (Ca9.54P5.98O23.8Cl1.60(OH)(2.74)). Both the sintered scaffolds displayed progressive deformation and delayed fracture under compressive loading, with hydroxyapatite-alginate scaffolds exhibiting a higher compressive strength (9.5 +/- 0.5MPa) than hydroxyapatite-maltodextrin scaffolds (7.0 +/- 0.6MPa). The difference in properties is explained in terms of the phase assemblage and microstructure.