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Articles by Kuldeep Sharma
Total Records ( 3 ) for Kuldeep Sharma
  Dharmender Singh , Yashpal Singh Malik , Kuldeep Sharma and Kuldeep Dhama
  In this study we performed the sequence and phylogenetic analysis of peste des petits ruminants (PPR) virus isolates recovered from natural disease outbreaks in goats of central India. A total of 58 clinical samples comprised of blood, nasal/oral/rectal swabs were collected from 3 natural PPR disease outbreaks. The samples were tested by using N protein based MAb based sandwich ELISA (sELISA) and subsequently RT-PCR targeting Fusion (F) and Nucleoprotein (N) genes of PPR virus. The F and N gene specific PCR amplicons were sequenced and assessed for sequence variations with existing and circulating PPR virus strains and vaccine isolates. The results of this study presents PPR virus antigen in 48.27% clinical samples (28/58) using sELISA confirming this outbreak of PPR in goats. Amplification with primers F1/F2 for F gene yielded an expected amplicon of 372 bp and N1/N2 primer sets yielded an amplicon of 463 bp for N gene in reference vaccine virus as well as 28 field samples. In phylogeny, different patterns of branching based on N and F gene sequences and clustering of many of the central Indian PPR virus isolates with isolates from bordering countries was seen. The results confirm that heterogeneous population of PPR virus isolates is circulating in India which moreover emphasize the significance of molecular methods to understand the epidemiology and diversity of PPR virus in the country. The information from such studies will help in achieving the target of controlling and eradicating the disease, especially when the country is planning to launch the control programs in many states after the successful stamping out of the Rinderpest.
  Kuldeep Sharma and G. Mahadevan
  Mobile ad hoc networks used for processing, storage and wireless communication capabilities of mobile devices to create unstructured and low-cost self-configuring networks. Black Hole Attacks are serious problems to establish a communication in MANETs. In this research, we present CCLAM, a new proposed approach using new classic clustering formatted graphs to identify the nodes who are trying to create a black hole. We use well-established architecture to gain information about the network topology, keep eyes on all ordinary nodes and master nodes and maintain database to update all kind of information which use to perform original checks of the routing information generated by the nodes in the network. We assume there is a node in the network generating fake and wrong routing information as malicious node. Therefore, we blow an flag and alarm if the verification check fails. Furthermore, we present promising first simulation results with the new architecture, it is possible to detect the attempt of creating a black hole before the actual attack occurs and prevent the network from malicious attack.
  Naveen Kumar , Yashpal Singh Malik , Kuldeep Sharma , Vinayagamurthy Balamurugan , Sathish Bhadravati Shivachandra and Kuldeep Dhama
  Rotaviruses of group A (RVA) are foremost cause of diarrhoeal diseases in neonates of animals and humans worldwide leading to substantial economic losses. The RVA non-structural protein-4 (NSP-4), a viral enterotoxin, is known to be associated with infantile gastroenteritis/secretory diarrhoea by inducing pathological changes in the mature enterocytes. In this study, the carboxyl terminus of NSP4 protein (73M to 175M) from a bovine RVA was expressed in Escherichia coli Tuner (DE3) pLysS cells. The fusion protein (rNSP4ct, ~31 kDa) with hexa-histidine tags on its both termini was purified by affinity chromatography under native condition using Nickel-Nitrilotriacetic acid (Ni-NTA) agarose resin. The purified soluble recombinant NSP4ct was confirmed by Western blot. The structural analysis of rNSP4 protein revealed similarity between bovine RVA and human RVA (central tetrameric coiled-coil region) and confirmed that it was composed of mainly alpha helix (85%), lacking the beta strands. The rNSP4ct protein of bovine RVA has the potential of being used in developing diagnostics, assessing the biological activity (enterotoxin property) of rNSP4ct in understanding the pathogenesis in intestinal mucosa which would reveal the role of anti-NSP4 antibodies in protection against rotavirus infection and stimulation of mucosal immunity in animal model.
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