Melanin Biosynthesis From Dopamine. II. A Mass Spectrometric and Collisional Spectroscopic Investigation

Electron impact (EI) and fast atom bombardment (FAB) mass spectrometry together with collisional activation (CA) experiments were applied to the study of the oxidation pathway of dopamine by tyrosinase. In order to prevent attachment of the protein to the highly reactive intermediates, ultrafiltration was employed to remove the enzyme at different reaction times. FAB, privileging molecular species formation, was successfully used for identification of transient intermediates and their relative concentrations with respect to time, directly in the reaction mixture. The presence of isobaric molecular species made chromatographic separation necessary. Further EI mass spectrometry and collision spectroscopy led to structural identification of pure components. Of these, dopamine-o-quinone, leucoaminochrome, and aminochrome semiquinone were characterized for the first time as real intermediates in dopamine melanogenesis, in agreement with previous hypotheses. This approach elucidated the pathway of dopamine melanogenesis.     

SENSITIVITY OF DAUDI CELLS TO TUMOR NECROSIS FACTOR ALPHA: EARLY CHANGES IN POLYPHOSPHOINOSITIDE METABOLISM

The effects of recombinant Tumor Necrosis Factor α (r-TNF α) on polyphosphoinositide metabolism were examined in a Burkitt Lymphoma cell line (Daudi cells). After 1h of in vitro treatment with r-TNF α, the incorporation of³²Pi into phosphatidylinositol 4,5-phosphate (PtdInsP₂), phosphatidylinositol 4-phosphate (PtdInsP) and phosphatidylinositol (PtdIns) was reduced compared with controls, confirming previous findings observed in other cell lines of a specific PtdIns breakdown following r-TNF α treatment. The novelty of this study is therefore the demonstration of early changes in polyphosphoinositide metabolism during the antiproliferative response elicited by this cytokine in Daudi cells.     

Strong correspondence between gillnet catch per unit effort and hydroacoustically derived fish biomass in stratified lakes

1. Sampling of lake fish assemblages is a challenging task in fish science, and the information obtained strongly depends on the choice of sampling gear. The use of more than one sampling technique is generally preferred in order to achieve a comprehensive view on fish assemblage structure. Therefore, the knowledge of whether catches between fishing gears are comparable is crucial. 2. We compared catches in benthic multi‐mesh gillnets with fish biomass estimates obtained by vertical hydroacoustics in 18 European lakes strongly varying in morphometry and trophic status. Separate analyses were conducted for different depth strata and for several fish length thresholds to account for depth‐ and size‐selective gillnet catches. 3. Gillnet catches and hydroacoustically obtained fish biomass estimates were significantly correlated. The strength of correlations was independent of the fish length thresholds applied, but varied across different depth strata of the lakes, with the strongest correlations occurring in the shallow strata. 4. The results support the applicability of vertical hydroacoustics for the quantification of fish biomass in stratified lakes. Survey designs combining hydroacoustics with limited gillnetting at sampling dates shortly one after the other, the latter for the purpose of inventory sampling only, are a cost‐effective strategy for sampling fish assemblages in lakes. However, gillnet sampling does not provide reliable fish density estimates in very deep lakes with separate, pelagic‐dwelling fish assemblages.