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Data from the article by Brodsky et al. (2000) have been examined in order to confirm that the fluctuations they report are not random but have a rhythmic basis, thereby verifying their conclusions regarding the existence of periodicity in protein synthesis. Reasons are outlined why oscillations should be expected in all cellular biochemical studies. Associated practical problems are briefly discussed together with comments on the validity and value of less rigorous methods of analysis. The subject of aliasing is raised as justification for doubting the validity of much published data, especially where periodicity has not been suspected. The existence of oscillations indicates the need for a thorough re-evaluation of our understanding of cell biochemistry.

Life cannot be simply defined in biochemical terms but it is associated with autodynamic behaviour. This fact implies that all aspects of cell biology should be viewed in terms of the resultant temporal features. Theoretical arguments indicate that the dynamic state can be explained only by the existence of periodicity. In accordance with this view, experimental evidence indicates the existence of multiple oscillators and at least some are highly complex, implying that failure to understand aspects of cell biology can stem from inadequate concepts.

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We used a system of co-culture of adult rat hepatocytes with another epithelial cell type from rat liver to study the synthesis of two acute-phase reactants, alpha-1 acid glycoprotein (alpha 1AGP) and the third component of complement (C3), and we have obtained long-term secretion of these two proteins. After a period of adaptation corresponding to the first 2-4 days of the co-culture, hepatocytes secreted C3 and alpha 1AGP for at least 2 weeks at a mean level higher than that observed in the first days of a pure culture of hepatocytes. When pulse-chase analysis was performed on day 6 of co-culture, kinetics of synthesis of alpha 1AGP and C3 were the same as those observed on day 1 of a conventional culture of pure hepatocytes. Furthermore, intracellular and extracellular alpha 1AGP had Mr values respectively of 39,000 and of 42,000-52,000, identical with those observed in pure cultures of hepatocytes. Similarly, the molecular size and subunit structures of C3 were the same in co-culture and in cultures, indicating an identical processing of this protein. C3 produced in co-culture was also haemolytically active. Therefore, the system of adult hepatocytes co-cultured with this liver epithelial cell provides a physiological system in vitro which permits long-term synthesis of the two acute-phase reactants C3 and alpha 1AGP. This model opens the possibility to study the modulation of the synthesis of these two proteins during a long period by inflammatory agents or by hormones.

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