Arginase from kiwifruit: properties and seasonal variation

The in vitro activity of arginase (EC 3.5.3.1) was investigated in youngest-mature leaves and roots (1-3 mm diameter) of kiwifruit vines (Actinidia deliciosa var. deliciosa) during an annual growth cycle, and enzyme from root material partially purified. No seasonal trend in the specific activity of arginase was observed in roots. Measurements in leaves, however, rose gradually during early growth and plateaued c. 17 weeks after budbreak. Changes in arginase activity were not correlated with changes in the concentration of arginine (substrate) or glutamine (likely end-product of arginine catabolism) in either tissue during the growth cycle. Purification was by (NH4)2SO4 precipitation and DEAE-cellulose chromatography. The kinetic properties of the enzyme, purified 60-fold over that in crude extracts, indicated a pH optimum of 8.8, and a Km (L-arginine) of 7.85 mM. Partially-purified enzyme was deactivated by dialysis against EDTA, and reactivated in the presence of Mn²⁺, Co²⁺, and Ni²⁺

Minimal in vivo efficacy of iminosugars in a lethal Ebola virus guinea pig model

The antiviral properties of iminosugars have been reported previously in vitro and in small animal models against Ebola virus (EBOV); however, their effects have not been tested in larger animal models such as guinea pigs. We tested the iminosugars N-butyl-deoxynojirimycin (NB-DNJ) and N-(9-methoxynonyl)-1deoxynojirimycin (MON-DNJ) for safety in uninfected animals, and for antiviral efficacy in animals infected with a lethal dose of guinea pig adapted EBOV. 1850 mg/kg/day NB-DNJ and 120 mg/kg/day MON-DNJ administered intravenously, three times daily, caused no adverse effects and were well tolerated. A pilot study treating infected animals three times within an 8 hour period was promising with 1 of 4 infected NB-DNJ treated animals surviving and the remaining three showing improved clinical signs. MON-DNJ showed no protective effects when EBOV-infected guinea pigs were treated. On histopathological examination, animals treated with NB-DNJ had reduced lesion severity in liver and spleen. However, a second study, in which NB-DNJ was administered at equally-spaced 8 hour intervals, could not confirm drug-associated benefits. Neither was any antiviral effect of iminosugars detected in an EBOV glycoprotein pseudotyped virus assay. Overall, this study provides evidence that NB-DNJ and MON-DNJ do not protect guinea pigs from a lethal EBOV-infection at the dose levels and regimens tested. However, the one surviving animal and signs of improvements in three animals of the NB-DNJ treated cohort could indicate that NB-DNJ at these levels may have a marginal beneficial effect. Future work could be focused on the development of more potent iminosugars