Effects on NaeI-DNA recognition of the leucine to lysine substitution that transforms restriction endonuclease NaeI to a topoisomerase: a model for restriction endonuclease evolution.

Substituting lysine for leucine at position 43 (L43K) transforms NaeI from restriction endonuclease to topoisomerase and makes NaeI hypersensitive to intercalative anticancer drugs. Here we investigated DNA recognition by Nael-L43K. Using DNA competition and gel retardation assays, NaeI-L43K showed reduced affinity for DNA substrate and the ability to bind both single- and double-stranded DNA with a definite preference for the former. Sedimentation studies showed that under native conditions NaeI-L43K, like NaeI, is a dimer. Introduction of mismatched bases into double-stranded DNA significantly increased that DNA's ability to inhibit NaeI-L43K. Wild-type NaeI showed no detectable binding of either single-stranded DNA or mismatched DNA over the concentration range studied. These results demonstrate that the L43K substitution caused a significant change in recognition specificity by NaeI and imply that NaeI-L43K's topoisomerase activity is related to its ability to bind single-stranded and distorted regions in DNA. A mechanism is proposed for the evolution of the NaeI restriction-modification system from a topoisomerase/ligase by a mutation that abolished religation activity and provided a needed change in DNA recognition.     

Phagocytosis activity of monocytes and macrophages in patients with plasmacytoma

The capacity to phagocytosis in monocytic macrophages was investigated in 24 patients with plasmocytoma. The investigation of this capacity was made by using short-term cultures with sheep erythrocytes loaded with antibodies and complement. In comparison to the control group the percentage of phagocyting macrophages was lowered in 16 from 24 patients. This impairment of the function of monocytic macrophages seems to be bound neither to the type of plasmocytoma nor to the stage of disease. Among other things, the nature of cell defect is discussed. The necessity of further investigations is emphasized.     

Microbial characteristics of sandy soils exposed to diazinon under laboratory conditions

Changes in microbiological characteristics in response to diazinon, applied at three different dosages of 7, 35 and 700 mg kg^?1 soil, were studied in pots filled with sandy soils of different texture. The insecticide dosages corresponded to the maximum predicted environmental concentration (PEC) in field conditions and five or hundred times this rate, respectively. To ascertain these changes, activities of selected soil enzymes, numbers of some microbial populations and bacterial community structure (r/K-strategists) were determined. Acid phosphatase activity and concentrations of ammonium ions either increased or were unaffected by the diazinon dosages. In contrast, nitrate ions, alkaline phosphatase, urease and especially dehydrogenase activities decreased in both soils treated with the higher dosages of the insecticide. The diazinon treatments increased the numbers of culturable bacteria and fungi; however, N-fixing bacteria and nitrifiers (but not denitrifiers) were decreased in both soils. When the high concentration of diazinon (100-times PEC) was added to the loamy sand soil (LS), simulating an undiluted chemical spill, the bacterial community structure shifted towards domination of K-strategists over time. However, r-strategists dominated in the community exposed to the same dosage of the insecticide in the sandy loam soil (SL). Generally, SL soil characteristics were more suitable for microbial activity and growth than those of LS soil were. The results indicate that except a situation of undiluted spill diazinon seems to be a non-hazardous chemical for the culturable soil microorganisms when applied at recommended concentrations.     

Sensitivity of insect ovarian tissues to various pteridines as tested in tissue culture

The effect of pteridine derivatives and analogues on the cell outgrowth from the ovarian explants of the waxmoth, Galleria mellonella, was examined in hanging-drop cultures and cytochemical tests were made for succinate and glucose-6-phosphate dehydrogenases. Most of the derivatives and analogues of 2-amino-4-hydroxypteridine injured to a lesser or greater extent insect ovarian tissues in vitro, depending on the drug structure and the concentration applied. Of all the pteridine derivatives and analogues tested only 2-amino-4-mercaptopteridine and its C-6,7-dimethyl derivative (10 μM) promoted cell outgrowth from the explants as did folate.