ABSTRACT
OBJECTIVE: With the help of bioinformatics analysis, we wished to develop a novel antivenom against the Deinagkistrodon (D.) acutus snake venom using B-cell linear epitopes of three primary toxins (serine protease, metalloprotease, and phospholipase A2). MATERIALS AND METHODS: cDNA sequences of three toxins of D. acutus venom were retrieved from the NCBI database. B-cell linear epitopes were predicted using DNAStar and the website server software provided by IEDB. Then, the sequences of the predicted epitopes were artificially synthesized and inserted into the vector pET-32a-c(+). Recombinant antigen peptide was expressed and purified. BALB/c mice were immunized with the recombinant antigen peptide. The immunoprotective effect of this novel antivenom was measured by neutralization of venom haemorrhagic activity. RESULTS: Six epitopes were obtained by bioinformatics analysis. ELISA analysis showed that antibody titre was >8,000 against snake venom and >64,000 against the recombinant peptide. Neutralization assays confirmed that the developed antivenom could effectively reduce the haemorrhagic activity of snake venom. CONCLUSIONS: Six B-cell linear epitopes of D. acutus snake venom were predicted by bioinformatics analysis and successfully utilized to produce a novel antivenom.
Subject(s)
Antivenins/genetics , Computational Biology/methods , Drug Design , Epitopes, B-Lymphocyte/genetics , Venoms/genetics , Amino Acid Sequence , Animals , Antivenins/therapeutic use , Base Sequence , Hemorrhage/chemically induced , Hemorrhage/prevention & control , Mice , Mice, Inbred BALB C , Molecular Sequence Data , Venoms/toxicityABSTRACT
One hundred forty three pairs of cancer patients and healthy controls were enrolled in this study. Thirty four psychologic stress factors were analyzed by stepwise and logistic regressions to identify risk factors for cancer. At the level of significance of alpha = 0.05, F(10,120) = 1.91 and alpha = 0.01, F(10,120) = 2.47, 15 and 13 risk factors were entered into stepwise regression equation, respectively. By conditioned logistic regression analysis, the following five major correlated factors were revealed: introversive personality, family accidents, the ten-year turmoil, repeated mishaps and death of children. The results indicate that, among all the psychological stress factors studied, introversive personality is the most important predisposing factor in the causation of cancer. This study suggests that psychologic stress might play important part in the etiopathogenesis of human malignancies.
