Comparison of different methods of determining cell viability after exposure to cytotoxic compounds

Cell-survival (of DON and L1210 cells) after treatment with cytotoxic compounds was assessed by measuring cloning efficiency, exclusion of trypan blue and erythrosin B, [⁵¹Cr] release, and attachment of DON cells to glass. Cell survival as measured by cloning efficiency did not correlate with survival measured by any of the other methods. We found that the stainability of cells after drug exposure depended on the cell line used. For example, after 3 h exposure to tubercidin although 100% of both DON and L1210 cells were killed (on basis of cloning efficiency), only 11% of DON cells and 68% of L1210 cells were dead as indicated by staining with erythrosin B. The stainability of cells also depended on the particular drug used. For example, after 24 h exposure of L1210 cells to adriamycin and tubercidin (both killed >99% of cells on basis of cloning efficiency) 21% of cells exposed to adriamycin and 99% of cells exposed to tubercidin were stained. The results obtained with several other cytotoxic compounds are discussed.     

The application ofin vivo techniques in the study of metabolic aspects of ion absorption in crop plants

Summary Little is known about the biochemical basis of the genotypic differences in the capacity for ion absorption and transport shown by many crop species. If these differences reflect the abundance of a specific membrane component or the activity of an enzyme we need to have some indication of thein vivo operation of these systems in whole plants. Thein vivo assessment of glycolytic enzymes is illustrated by the effects of mannose on the transport of phosphate in maize varieties. The application of high resolution³¹P-NMR to the study of intermediary metabolismin vivo is also helpful in following transport capacity. The five-fold rise in respiratory rate that occurs when freshly cut potato slices are maintained in aerated water for 24 hours is accompained by the turning on of a wide range of biochemical systems. Major increases in the capacity for absorption of phosphate from low concentrations (0.1 μM–10 μM) and in the phosphorylative ability of the tissue are seen, indicating the synthesis of a carrier involved in phosphate transport. These capacities differ markedly between individual tissues of the tuber,i. e. pith, parenchyma, cortex and buds and large differences have been observed between comparable tissue from different varieties. Varieties grown under similar conditions have been compared and shown to exhibit different kinetics with respect to the development of the low concentration absorption site and in their sensitivity to the effects of uncouplers such as 2,4-dinitrophenol.     

Nitrogen assimilation in field-grown winter wheat: Direct measurements of nitrate reduction in roots using15N

Summary Nitrate fertiliser labelled with¹⁵N was applied to a field grown crop of winter wheat. Uptake and assimilation of fertiliser nitrate was studied by monitoring the appearance of labelled nitrate and labelled amino acids in the xylem sap. Shortly after applying¹⁵N-nitrate to the soil about 30 per cent of recently absorbed¹⁵N was in the reduced form, indicating that roots of cereal crops can make a substantial contribution in reducing nitrate. Seasonal changes in crop growth andin vivo NRA are also described.     

Effects of vanadate on the ATP content,ATPase activity and phosphate absorption capacity of maize roots

Although the sensitivity of the plasma membrane H⁺-ATPase to vanadate is well known, the metabolic response of plant cells to vanadate is less well characterised in vivo and its use as an inhibitor in whole plant experiments has had mixed success. Experiments with maize (Zea mays, L.) roots and with purified plasma membrane fractions from the same tissues showed that exposure to vanadate caused: (i) a reduction in the capacity for phosphate uptake; (ii) a reduction in the extractable ATPase activity from the tissue; and (iii) a significant increase in the ATP level. The measurements on the extractable ATPase activity and the ATP level showed that the effect of vanadate developed slowly, apparently reflecting the slow accumulation of intracellular vanadate. The marked effect of vanadate on the ATP level-exposure to 500 M vanadate for 5 h doubled the ATP content of the roots tips-indicates that there is no stringent control over the ATP level in the roots and that the plasma membrane H⁺-ATPase activity is likely to have a significant role in determining the ATP level under normal conditions.     

Current Status of Cereal Root Diseases in Western Australia Under Intensive Cereal Production and Their Comparison with the Historical Survey Conducted During 1976–1982

Two separate surveys of root diseases of cereals in the Western Australian (WA) cereal belt were conducted: the first conducted annually for wheat and barley during 1976–1982 and the second for wheat during 2005–2007. For the 1976–1982 survey, the cereal belt was divided into 15 zones based on the location and rainfall. Sampling was representative of the actual cropping area, with both wheat and barley sampling sites selected by zone as a percentage of total sites. Over 31 000 plants were assessed from a total of 996 fields. Average take‐all incidence ranged from 3% in the northern low rainfall zone to 57% in the southern high rainfall zone. Other root diseases assessed included rhizoctonia root rot, fusarium crown rot and subcrown internode discolouration. During the 2005–2007 survey, around 20 000 plants from a total of 210 fields being intensively cropped with cereals were surveyed for take‐all, rhizoctonia root rot, fusarium crown rot, common root rot, root lesion nematode and cereal cyst nematode. The 2005–2007 survey results indicated that root and crown diseases prevailed in paddocks frequently cropped with cereals and occurred at damaging levels across all WA cropping districts surveyed. The more recent root disease survey identified that the fungal diseases rhizoctonia root rot and fusarium crown rot and the root lesion nematode were the most serious impediments to intensive cereal production, particularly in the southern region of WA. Comparing the 2005–2007 results with the previous survey of 1976–1982, the relative importance of take‐all appears to have declined over the past 30 years.     

Clumping factor A of Staphylococcus aureus inhibits phagocytosis by human polymorphonuclear leucocytes

Staphylococcus aureus is a major cause of nosocomial and community-acquired infection. It expresses several factors that promote avoidance of phagocytosis by polymorphonuclear leucocytes. Clumping factor A (ClfA) is a fibrinogen-binding surface protein of S. aureus that is an important virulence factor in several infection models. This study investigated whether ClfA is an antiphagocytic factor, and whether its antiphagocytic properties were based on its ability to bind fibrinogen. In S. aureus, ClfA was shown to be of equal importance to protein A, the antiphagocytic properties of which are well established. ClfA expressed in a surrogate Gram-positive host was also found to be antiphagocytic. A ClfA mutant that was unable to bind fibrinogen had a similar antiphagocytic effect to native ClfA in the absence of fibrinogen. ClfA inhibited phagocytosis in the absence of fibrinogen, and showed enhanced inhibition in the presence of fibrinogen.     

Studies of the immunological capacity of germ-free mouse radiation chimeras. II. Thymus cell function in a humoral immune response to sheep erythrocytes

Experiments described here were undertaken to determine the reason for the depressed humoral immune response in germ-free mouse allogeneic radiation chimeras. Indirect immunofluorescence using the theta (θ) antigen as a marker demonstrated that about 10% of the nucleated cells in the spleen of both allogeneic and syngeneic chimeras bear the θ antigen. One type of in vivo cell transfer assay employed to determine the capacity for “helper” function of thymocytes revealed that allogeneic chimera thymocytes were only 7–18% as efficient in “helper” function as normal thymocytes. A second type of in vivo cell transfer assay demonstrated that the presence of intact normal thymic stroma had no effect on the “helper” inefficiency of thymocytes obtained from allogeneic radiation chimeras.