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The purpose of the study was to estimate the influence of temperature increase on Hsp70 induction in M. haemolytica serovar 1 strains. Three wildtype M. haemolytica strains, obtained from calves respiratory tracts and incubated at a temp of 41.5°C for 2 hours were used as the research material. Analyses of particular fractions were carried out by SDS-PAGE electrophoresis and identification of obtained proteins by immunoblotting (Western blotting) using polyclonal rabbit anti Hsp70 antibodies. The first step was to separate the capillaries in gradient pH 5/7 and 3/10 which was carried out in two-dimensional electrophoresis. The second step was carried out in SDS-PAGE electrophoresis using 4% stocking and 12% resolving gels. An analysis of SDS-PAGE electrophoresis revealed additional protein fractions, displaying positive reactions with anti-Hsp70 antibodies. The presence of these proteins was observed both in membrane and cytoplasmatic bacterial cell fractions. The molecular weight of the obtained proteins ranged between 77.5-79 kDa. The additional protein fractions were present in membrane fractions between molecules, weighed 22-26 kDa, as well as displaying a positive reaction with anti-Hsp70 antibodies. The electrophoregrams obtained in 2D electrophoresis revealed the presence of additional spots in membrane, cyto- and periplasmatic fractions. The obtained results suggest the potential for M. haemolytica strains to produce Hsp70 during stress induced by temperature increase.
Ectromelia virus (ECTV) is a member of the Orthopoxvirus genus of the Poxviridae family. It is a causative agent of mouse pox in genetically sensitive mice strains of H-2ᵈ (e.g. BALB/c) and H-2ᵃ (eg.A, A/J) haplotypes. Mouse pox virus is a well recognized model for studying generalized viral infections in natural hosts. The aim of this study was to determine the role of heat shock proteins (namely hsp90, hsp70 and hsp2) during the replication of the Moscow strain of ECTV (ECTV-MOS) in BALB/c mice. The internal organs of mice are important sites for primary virus replication whereas ECTV penetration into the brain may seriously influence mechanisms supervising immune and nervous systems cooperation. Based on studies carried out in BALB/c mice infected with ECTV-MOS, it was found that the hsp (hsp90, hsp70 and hsp27) expression in brain cells reach peak values during the incubation period and clinical manifestation of mouse pox but the relative numbers of hsp⁺ cells in the brains decreased during the recrudescence of the infection to values observed in the control mice. The high expression of hsp (hsp90, hsp70 and hsp27) on oligodendrocytes in BALB/c mice during the incubation and clinical periods may reflect the protective action of heat shock proteins within the brain.
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