Bioaccessibility of Cd, Cu, Fe, Mn, Pb, and Zn in hazelnut and walnut kernels investigated by an enzymolysis approach.

Bioaccessibility of four essential (Fe, Cu, Mn, Zn) and two toxic (Cd, Pb) elements in kernels of four walnut and four hazelnut cultivars was investigated using sequential enzymolysis approach and atomic absorption spectrometry. It was found that the assimilable part of elements was not dependent on nut cultivar. The bioaccessible fraction of Cu, Mn, and Zn was definitely higher for hazelnuts (62% Cu, 39% Mn, 58% Zn) than for walnuts (14% Cu, 21% Mn, 15% Zn). Bioaccessible Fe was 20-24% from its total content for both nut types. Solubility in the simulated intestinal juice is affected by both formation of stable soluble complexes and back sorption of dissolved elements on nut solid residues. Lead shows strong insolubility due to the high sorption affinity of lead ions to the insoluble fraction of nuts. Thus, walnuts and hazelnuts could act as effective biosorbents for lead detoxication.

Cytokinin N-glucosylation inhibitors suppress deactivation of exogenous cytokinins in radish, but their effect on active endogenous cytokinins is counteracted by other regulatory mechanisms.

The prolongation of the effect of exogenous cytokinins by inhibition of their inactivation was studied in Raphanus sativus L. cv. Rampouch. As in radish the main way of cytokinin inactivation is their N-glucosylation, inhibitors of this pathway, papaverine, theophylline and olomoucine, as well as two olomoucine analogues, bohemine and roscovitine, were tested. The latter ones, which function as potent inhibitors of cyclin-dependent kinases, have also been found to effectively inhibit cytokinin N-glucosylation. Incubation (24 h) of de-rooted radish seedlings with inhibitors resulted in c. 50% decrease of the conversion of model cytokinins, [(3)H]dihydrozeatin and [(3)H]N(6)-benzyladenine, to the corresponding 7N-glucosides. Simultaneously the level of the non-metabolized cytokinin bases was elevated. The activity of cytokinin oxidase/dehydrogenase (EC 1.5.99.12) was suppressed in a dose dependent manner. The concentration of physiologically active endogenous cytokinins was not increased significantly by inhibitor application. The inhibition of N-glucosylation was in the case of olomoucine, bohemine and roscovitine accompanied by the accumulation of physiologically nonactive cis-zeatin derivatives. The impact of inhibitors on the endogenous cytokinin pool seems to be balanced by the mechanisms involved in the maintenance of cytokinin homeostasis.

Cyclin-dependent kinase inhibitor, roscovitine, in combination with exogenous cytokinin, N6-benzyladenine, causes increase of cis-cytokinins in immobilized tobacco cells.

Dynamics of the response of tobacco cells (line BY-2) to exogenous cytokinin, N6-benzyladenine, and cyclin-dependent kinase inhibitor, roscovitine, was followed using alginate-immobilized cells packed into a column. N6-Benzyladenine (1.25 microM) increased the synthesis of the physiologically-active endogenous cytokinin, isopentenyladenosine, in the effluent up to 0.1 nM. Simultaneously, conversion of the excess of endogenous cytokinins to biologically inactive derivatives of cis-zeatin occurred, up to 0.8 nM. Roscovitine (50 microM) further increased cis-cytokinins, up to 2.2 nM.