Faculty Bibliography

The possibility that serum ferritin is a secreted protein and an acute phase reactant regulated by inflammatory hormones and iron was examined in a hepatic cell line that secretes plasma proteins. Differentiated rat hepatoma cells released albumin and ferritin into the medium, as determined by rocket immunoelectrophoresis and isolation of ferritin by standard procedures plus immunoaffinity chromatography, following labeling with radioactive amino acid. Administration of interleukin-1-beta (IL-1) or tumor necrosis factor-alpha (TNF) doubled the amounts of ferritin released into the medium over 24 and 48 hours. Together, the cytokines had more than an additive effect. Albumin secretion was diminished by IL-1, but not TNF. Iron, administered as an iron dextran complex or as a 1:1 chelate with nitrilotriacetate (Fe-NTA), also enhanced ferritin release, but had no effect on albumin. Intracellular ferritin concentrations did not change significantly with cytokine treatment, but increased in response to iron. With or without treatments, release of ferritin and albumin from cells into the medium was inhibited by brefeldin A, an inhibitor of Golgi function. The effect of each of the cytokines and of iron on ferritin and albumin was also blocked by dichlorofuranosylbenzimidazole (DRB), an inhibitor of transcription. The stimulatory effect of Fe-NTA on ferritin secretion was diminished by TNF, and this was partially counteracted by IL-1, indicating additional regulatory complexity. These results show for the first time that hepatic cells secrete ferritin, that this ferritin secretion is regulated by iron and inflammatory cytokines, and that the mechanisms of regulation differ from those for intracellular ferritin. The results would explain why serum ferritin increases in inflammation or when iron flux is enhanced.

Serum ferritin isolated from the horse was structurally compared with horse spleen ferritin and was found to differ markedly in molecular weight, iron content, carbohydrate, subunit size and amino acid sequence. The results are summarized and initial results obtained with candidate clones of pieces of two serum ferritin subunits are described.

Heart rate variation was studied in 56 healthy subjects from ages 20-81 while supine and standing during spontaneous and metronome breathing. Time domain analyses revealed no age effect on supine heart rate but standing heart rate decreased with aging (p less than .008). The SD of R-R intervals decreased with increasing age in both positions (p less than .001). Frequency domain analysis (Fast Fourier Transformation of R-R interval data) showed total spectral content to decrease with aging (p less than .001). Both high frequency (0.20-0.32 Hz) and low frequency (0.04-0.12 Hz) content were reduced with aging (p less than .001). The ratio of high to low frequency content, however, was not affected by age in the supine position nor were changes in this ratio in response to standing during spontaneous breathing. Although the absolute increase in low frequency content upon standing was less with aging (p less than .001), the percent increase in low frequency content was not related to age. Metronome breathing decreased total spectral content (p less than .001) but increased high frequency content, especially in younger subjects (p less than .03). In response to standing, greater proportional and absolute decreases in high frequency content occurred in younger subjects resulting in age differences in the changes in ratios of high to low frequency content with standing during metronome breathing. In summary, decreased heart rate variation was seen with aging, but the balance between parasympathetic and beta-adrenergic input at rest and in response to postural changes does not appear to be influenced by age during spontaneous breathing. Metronome breathing altered spectral content within subjects and produced age-related differences in responses to postural maneuvers not seen during spontaneous breathing.