Studies on shipping fever of cattle. I. The experimental exposure of cattle with various cultures of Pasteurella.
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Hybridomas were produced which secrete antibodies against cell surface antigens of neoplastic bovine lymphocytes. The monoclonal antibodies of three hybridomas react only with leukemic cells but not with other tested cellular and soluble antigens. Four hybridomas secrete antibodies which probably recognize virus-encoded structures on outer membranes of leukemic cells.
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Using cell extract fractionation with ammonium sulfate and subsequent chromatography on DEAE- and DNA-cellulose and Blue Sepharose, two cytosine DNA-methylases were isolated from blood lymphocytes of cows suffering from lympholeukosis; one cytosine DNA-methylase was isolated from blood lymphocytes of healthy animals. The DNA-methylases from normal lymphocytes was purified 383-fold; the enzyme has a specific activity of 2.3 u./mg, Mr of 114 000 Da and a pH optimum of 7.6. The molecular mass of DNA-methylases from leukemic lymphocytes is about 130,000 Da. The enzymes isolated from leukemic lymphocytes, i.e., DNA-methylase I (568-fold purification, specific activity 14.2 u./mg) and DNA-methylase II (524-fold purification, specific activity 13.1 u./mg) possess different action optima at pH 7.8 and 6.7, respectively. The total DNA-methylase activity of leukemic lymphocytes is about 4 times that of normal lymphocytes. All the DNA-methylases under study methylate in vitro bacterial and animal DNAs of different base composition; bacterial DNAs are the best (GC content is approximately 70%), while homologous DNAs- the worst acceptors of CH3-groups. Heat-denaturated DNAs are methylated more intensively than initial DNAs. The optimal NaCl concentration in the reaction mixture is approximately 50 mM; EDTA (greater than 10 mM) inhibits the reaction. DNA-methylase I of leukemic and normal lymphocytes show the same pH optimum and specificity of action. In vitro methylation of bacterial DNA by these DNA-methylases in the presence of [3H-methyl]S-adenosylmethionine results in the similar type of label distribution among pyrimidine isopliths in the DNA. DNA-methylase II from leukemic lymphocytes methylates about two times more Pu-C-Pu sequences in th same DNA than does DNA-methylase I from leukemic and normal lymphocytes and thus reveals a different specificity. The changes in the type of DNA methylation as well as the appearance of an additional DNA-methylase possessing a different specificity of action in leukemic lymphocytes may be responsible for cell transformation and transcriptional changes in chronic lympholeukosis.
Magnesium (Mg) concentrations in plasma, serum, and erythrocytes (RBC) of calves subjected to splenectomy (SP), splenectomy-induced eperythrozoonosis (EP), and experimentally induced anaplasmosis (AN) were determined and compared with pre-SP, pre-EP, and pre-AN values. Mean serum and plasma Mg concentrations varied in parallel through most periods of the trial. Both were highest after SP, but neither value was significantly different from the pre-SP concentrations. Mean serum and plasma Mg values varied inversely with mean RBC values after SP and EP. Mean RBC Mg values increased significantly from previous periods during EP and AN and after AN, with the values after AN being significantly higher than those at all other periods. Mean plasma, serum, and RBC Mg concentrations were correlated to both mean percentage of parasitemia (%P) and packed-cell volume (PCV). The RBC Mg values increased significantly after mean %P reached 20 and the PCV began to decrease. There was evidence for a distinct lag variation in increasing RBC Mg concentrations with respect to both mean %P and PCV during AN. A strong correlation was observed between the RBC Mg values and the %P occurring 5 days previously and the PCV occurring 3 days previously.
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