Dissociation between murine spleen cell mitogenic activity of enterotoxin contaminants and anti-tumor activity of staphylococcal protein A

Soluble staphylococcal protein A (SpA) in the form of high m.w. complexes with IgG has been shown to significantly inhibit the growth of Meth A fibrosarcomas in BALB/c mice. Although SpA reportedly is a potent T cell mitogen that can induce immune cell proliferation and production of humoral factors with anti-tumor activity, it has been suggested that mitogenic enterotoxin contaminants might be responsible for these effects. The purpose of the present study was to investigate the nature of SpA-induced cell proliferation and the relationship between mitogenicity and the anti-tumor effect that we observed in our mouse model. SpA stimulated the proliferation of a mixed population of splenic B and T cells from BALB/c mice, but activity did not require the presence of IgG in the culture medium. Furthermore, mitogenic activity could be inhibited completely by anti-SEA plus anti-SEB, but was unaffected by anti-SpA. HPLC-purified SpA was inactive while the mitogenic factor(s) had the same retention time as authentic enterotoxin and its activity was inhibited by anti-SEA and anti-SEB, but not by anti-SpA. Enterotoxin-free rSpA produced in Escherichia coli had the same IgG binding capacity as the staphylococcal product but was not mitogenic. These data indicate that SEA and SEB completely account for mitogenicity in SpA preparations. In contrast, we found that optimal concentrations of rSpA as well as crude and HPLC purified staphylococcal SpA were equally effective in inhibiting the growth of established Meth A fibrosarcomas demonstrating that SpA is responsible for antitumor activity without any apparent role for enterotoxins.     

Feeding ecology of common carp (Cyprinus carpio L.) in a rice–fish culture system of the Apatani plateau (Arunachal Pradesh, India)

For two years (2002, 2003) selective feeding ecology of the common carp (Cyprinus carpio L.) has been studied in carp-integrated rice fields in Apatani Plateau of Arunachal Pradesh (India). Sampling strategy was based on the water depths in the fields and on the flood phases: early flood phase (June–July), mid flood phase (July–August), and late flood phase (September–October). In 2003 the water level was higher and therefore periphyton availability was better. This resulted in larger gut contents and better growth of the carp compared with 2002 when the water levels were lower. Gut contents analyses revealed a total of 60 food items of which 22 belonged to the Chlorophycea, 12 to the Cyanobacteria, 10 to the Bacillariophycea and 16 to several zooplankton taxa. With the progress of flood phases, the fish increased its feeding on periphyton food items; simultaneously, feeding on plankton items gradually declined. This was caused by the increasing periphyton availability on the rice-stems. Selective feeding on plankton and periphyton taxa was studied, selectivity changed with the flood phases. Periphytic Chlorophycea and Cyanobacteria, especially, were strongly positively selected. Generally, periphyton was the most important resource for the common carp in the rice fields.     

Influence of duodenal secretions and its components on release and activities of human brush-border enzymes

The in vitro effects of human duodenal secretions and various combinations of its components on activity and release of enzymes from the human brush border were examined. Sucrase retained activity for 90 min in duodenal secretions, and maltase was almost as stable; lactase lost activity rapidly and alkaline phosphatase was of intermediate stability. Inactivation of lactase could only be partly (50%) attributed to luminal proteases, bile salts and phospholipids played no role. Rate of release of an enzyme from the brush border bore no relationship to its rate of inactivation. When individual proteases were studied, elastase was the most potent for releasing disaccharidases from the brush border; trypsin was ineffective alone but augmented the effect of elastase. Sucrase and maltase were activated by proteolytic release, but activation was abolished by simultaneous exposure of brush borders to bile salts. Lactase was released and rapidly inactivated by proteinases, while alkaline phosphatase appeared to be inactivated without significant release. These results show that there are significant interactions between luminal factors which have been inapparent when studying them in isolation. Loss of functionally useful enzyme does not follow release of sucrase or maltase from the brush border into the lumen but does follow release of lactase. Study of the susceptibility of lactase to inactivation by luminal factors in the various forms of lactose intolerance is warranted.     

A simplified method for micronucleus preparation from hepatic cells

A simplified method for micronucleus preparation from regenerating hepatocytes has been developed. Small pieces of the regenerating portion of the liver are incubated in 1% sodium citrate solution containing collagenase Type 1A (final concentration 0.005% w/v) at 37 C for 10-15 min with occasional gentle agitation. The larger particles are discarded. Drops of the thick homogeneous citrate suspension of liver cells are put on the slides and drawn back immediately into the pipette, leaving only the drop marks. This simplified method, which gives good preparations with many intact hepatocytes, was validated in a model experiment using mitomycin C. The data revealed a distinct dose-response effect.