A new imagination for waste and water in India’s peri-urban interface

Cities are often seen as incubators for enterprise and innovation. However, in this urbanization era, we seem to suffer from a lack of imagination on how to handle the many environmental problems associated with expanding cities. This is especially true in the case of the peri-urban interface (PUI), a geographical and conceptual landscape with which the city core often has a contentious relationship. In this chapter, we look at the complex linkages between water and waste in the PUIs of two metropolitan cities: Bengaluru and Kolkata. We look at two water systems: Kannuru lake in Bengaluru and Kolkata’s wetlands. Kannuru is a freshwater lake that supported traditional livelihoods and subsistence use by local communities, while Kolkata’s peri-urban wetlands not only served as the city’s natural sewage treatment plant but also enabled agriculture and aquaculture. Urbanization has adversely impacted both these water systems. Kannuru lake is threatened by a landfill on its periphery, while sewage-based farming and fisheries in Kolkata’s wetlands have been impacted by changes in land use and composition of sewage. We unravel the complexity in the waste-water relationship, where waste is seen as a pollutant in one and as a nutrient in the other. We attempt to understand how we can re-envision waste and water linkages in the PUIs of expanding cities if India needs to move towards a sustainable future

Processing of multimer FMD virus VP1-2A protein expressed in <i style="">E. coli</i> into monomers

760-763Expressions of several genes in bacteria were carried out by independent promoter. However, in case of eukaryotes ribosome skipping and introduction of IRES are employed as alternative to multiple translation initiation. Foot and mouth disease virus (FMDV) 2A peptide has been widely used for co-expression of multiple genes in eukaryotic, plant and mammalian systems. The 18 amino acid 2A peptide of FMDV facilitates efficient co-translational dissociation of the polyprotein into discrete protein products. To study the role of 2A in multimeric protein production a construct consisting of tandem repeat of 4 units of C- terminal VP1 linked through 2A sequence was made and expressed in E. coli. Along with tetramer protein, trimer, dimer and monomer proteins were produced. Stability studies showed that the tetramer protein was cleaved to smaller monomer on storage. The results provide scope for using FMDV 2A for expressing multiple genes under a single promoter in prokaryotes

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Not AvailableRNA interference (RNAi) has been used as an effective antiviral strategy for its specific silencing of viral gene expression in mammalian cells. In this study, shRNA targeting two regions of Foot and Mouth Disease Virus (FMDV) i.e. 3D and 5′UTR which are very essential in virus replication were evaluated. The constructs were made using h7K RNA polymerase III promoter. We investigated in vivo inhibitory effect of shRNA on FMDV replication in BHK-21 cells and guinea pigs. The results showed that transfection of 3D shRNA could reduce virus growth by three folds when cells were challenged with 102 TCID50 of FMDV. Pretreated guinea pigs with 3DshRNA were protected 80% with 103 GPID50 of FMDV. As a first report in guinea pigs which are recognized animal model for FMD vaccine potency testing, the study suggests that shRNA could be a viable therapeutic approach to control severity of FMD infection and spread.Not Availabl

Cloning and expression of FMDV-VP1 immunoreactive peptide in trivalent form and its application as immunogen

472-477A search for alternatives to conventional inactivated virus vaccine for FMD with an aim to control and eradicate the disease globally, is a continuous process till a promising one is identified. Development of such vaccines underlines necessity of avoiding the use of active virus, and to have broader antigenic coverage so as to make them suitable even for disease free countries. Subunit or peptide vaccines have been shown to elicit neutralizing antibody response. However, the titres are low as compared to sera from animals vaccinated with conventional vaccine and fail to protect animals against virus challenge. This is probably due to the inclusion of only limited epitopes. Under such conditions, mixing heterologous epitopes from the various serotypes may be a better approach for elicitation of high titred antibody response. Keeping this in view, we have linked C-terminal half of VP1 carrying two B cell and one T cell epitope of three FMDV serotypes (O, A and Asia 1), which are presently in use as vaccine strains in India. The linked polyvalent gene was expressed in Escherichia coli and the 59 kDa fusion protein was studied for its immunogenicity in guinea pigs in comparison with the specific epitopes of type ‘O’ produced as a similar fusion protein of 30 kDa. The trivalent protein showed better neutralizing antibody response, even with single booster injection, as compared to monovalent protein as observed in ELISA and SNT. These studies show future scope for the development of protein/DNA-based vaccine for FMD

Amplification, cloning and sequencing of Enterococcus feacalis enolase gene

307-312α-Enolase, a key glycolytic enzyme, belongs to a novel class of surface proteins, which do not possess classical machinery for surface transport and transported on the cell surface through an unknown mechanism. It is a multifunctional protein and its ability to serve as a plasminogen receptor on the surface of a variety of hematopoetic, epithelial and endothelial cells suggest that it may play an important role in the intravascular and pericellular fibrinolytic system. Authors have amplified and cloned α-enolase gene of Enterococcus feacalis in a prokaryotic cloning vector, and then transferred it into Escherichia coli. The recombinant enolase vector (r-pBEnol) was isolated and sequenced. The sequence of the cloned enolase from E. feacalis was found identical to that of the E. feacalis V583. The sequence was submitted to NCBI nucleotide data bank and accession number (AM279410) was obtained