Deepshikha Mishra

Characterization of a novel human sperm-associated antigen 9 (SPAG9) having structural homology with c-Jun N-terminal kinase-interacting protein.

We report a novel SPAG9 (sperm-associated antigen 9) protein having structural homology with JNK (c-Jun N-terminal kinase)-interacting protein 3. SPAG9, a single copy gene mapped to the human chromosome 17q21.33 syntenic with location of mouse chromosome 11, was earlier shown to be expressed exclusively in testis [Shankar, Mohapatra and Suri (1998) Biochem. Biophys. Res. Commun. 243, 561-565]. The SPAG9 amino acid sequence analysis revealed identity with the JNK-binding domain and predicted coiled-coil, leucine zipper and transmembrane domains. The secondary structure analysis predicted an alpha-helical structure for SPAG9 that was confirmed by CD spectra. Microsequencing of higher-order aggregates of recombinant SPAG9 by tandem MS confirmed the amino acid sequence and mono atomic mass of 83.9 kDa. Transient expression of SPAG9 and its deletion mutants revealed that both leucine zipper with extended coiled-coil domains and transmembrane domain of SPAG9 were essential for dimerization and proper localization. Studies of MAPK (mitogenactivated protein kinase) interactions demonstrated that SPAG9 interacted with higher binding affinity to JNK3 and JNK2 compared with JNK1. No interaction was observed with p38alpha or extracellular-signal-regulated kinase pathways. Polyclonal antibodies raised against recombinant SPAG9 recognized native protein in human sperm extracts and localized specifically on the acrosomal compartment of intact human spermatozoa. Acrosome-reacted spermatozoa demonstrated SPAG9 immunofluorescence, indicating its retention on the equatorial segment after the acrosome reaction. Further, anti-SPAG9 antibodies inhibited the binding of human spermatozoa to intact human oocytes as well as to matched hemizona. This is the first report of sperm-associated JNK-binding protein that may have a role in spermatozoa-egg interaction.

SECURITY OF COLOR IMAGE DATA DESIGNED BY PUBLIC-KEY CRYPTOSYSTEM ASSOCIATED WITH 2D-DWT

In present times the security of image data is a major issue. So, we have proposed a novel technique for security of color image data by public-key cryptosystem or asymmetric cryptosystem. In this technique, we have developed security of color image data using RSA (Rivest-Shamir-Adleman) cryptosystem with two-dimensional discrete wavelet transform (2D-DWT). Earlier proposed schemes for security of color images designed on the basis of keys, but this approach provides security of color images with the help of keys and correct arrangement of RSA parameters. If the attacker knows about exact keys, but has no information of exact arrangement of RSA parameters, then the original information cannot be recovered from the encrypted data. Computer simulation based on standard example is critically examining the behavior of the proposed technique. Security analysis and a detailed comparison between earlier developed schemes for security of color images and proposed technique are also mentioned for the robustness of the cryptosystem.

Small interference RNA-mediated knockdown of sperm associated antigen 9 having structural homology with c-Jun N-terminal kinase-interacting protein

Abstract Recently, we reported a novel testis-specific sperm associated antigen 9 (SPAG9) protein, a new member of the JNK-interacting protein family, having a functional role in sperm–egg fusion [N. Jagadish, R. Rana, R. Selvi, D. Mishra, M. Garg, S. Yadav, J.C. Herr, K. Okumura, A. Hasegawa, K. Koyama, A. Suri, Biochem. J. 389 (2005) 73–82]. NCBI Blast searches revealed SPAG9 nucleotide sequence similarities with ESTs of various cancerous tissues. In the present study, we compared the efficiency of two independent SPAG9 specific small interfering RNA (siRNA) constructs, BS/U6/spag9 and BS/U6/spag9-I, to ablate the SPAG9 expression in mammalian cells. A positive correlation between the ratio of target gene versus siRNA and the suppression of SPAG9 expression was observed. Further, the cotransfection of BS/U6/spag9 with pcDNA-SPAG9 and pFlag-CMV2-JNK-3 resulted in specific suppression of SPAG9 without affecting JNK-3 expression. The present investigation will eventually extend the application of SPAG9 siRNA in in vivo targeting experiments that aim to define the SPAG9 functional genomics in tumor and reproductive biology.

TWO LAYERS OF SECURITY FOR COLOR VIDEO BY MATRIX AFFINE CIPHER WITH TWO-DIMENSIONAL DISCRETE WAVELET TRANSFORM

In this cryptosystem, we have presented a novel technique for security of video data by using matrix affine cipher (MAC) combined with two-dimensional discrete wavelet transform (2D-DWT). Existing schemes for security of video data provides only one layer of security, but the presented technique provides two layers of security for video data. In this cryptosystem, keys and arrangement of MAC parameters are imperative for decryption process. In this cryptosystem, if the attacker knows about all the exact keys, but has no information about the specific arrangement of MAC parameters, then the information of original video cannot be recovered from the encrypted video. Experimental results on standard examples support to the robustness and appropriateness of the presented cryptosystem of video encryption and decryption. The statistical analysis of the experimental results based on standard examples critically examine the behavior of the proposed technique. Comparison between existing schemes for security of video with the presented cryptosystem is also provided for the robustness of the proposed cryptosystem.