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Articles by H. Das
Total Records ( 1 ) for H. Das
  H. Das , S.U. Ahmed , S.K. Shukla , S. Shukla , A. Latif and D. Sharma
  Mastitis represents a failure of innate immune mechanism of udder teat canal epithelial cells and peripheral PMN cells, whose cationic peptides constituted innate immunity. An attempt was made to examine in the year 2007, the actual presence of β-defensins peptides in milk of mastitis-affected buffalo (Bubalus bubalis), due to lack of such information. AU-PAGE revealed higher frequency and density of the peptides in case of mastitis milk as compared to normal milk sample, particularly in the most cationic zone with highest electrophoretic mobility or lower molecular weight region. These low molecular peptides were also separated out using 10 kDa cut-off membrane ultra filtration, which was subsequently examined for in vitro antibacterial sensitivity and subjected to SDS-PAGE and low molecular weight zone was further subjected to MALDI-TOF mass spectrometry, which identified the most anodic two peptides of about 5 kDa as β-defensin viz. The LAP (lingual antibiotic peptide) and BNBD-2 (bovine neutrophil beta-defensin-2) based on amino acid sequences unlike that of healthy buffalo milk. Sequence homology studies also supported mass spectrometry data as both the peptides shared 100 and 98.5% identities with cattle β-defensins, respectively. Antibacterial assay against S. aureus and E. coli revealed significant activity of these peptides against both gram-positive and gram-negative organisms at 10 μg concentration. It is concluded that expression of β-defensin peptides in ductal epithelium were induced in mastitis and these defensin peptides were also released into the milk.
 
 
 
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