Intracellular thiol-disulfide redox balance is crucial to cell health, and may be a key determinant of a cancer's response to chemotherapy and radiation therapy. The ability to assess intracellular thiol-disulfide balance may thus be useful not only in predicting responsiveness of cancers to therapy, but in assessing predisposition to disease. Assays of thiols in biology have relied on colorimetry or fluorimetry, both of which require UV-visible photons, which do not penetrate the body. Low-frequency electron paramagnetic resonance imaging (EPRI) is an emerging magnetic imaging technique that uses radio waves, which penetrate the body well. Therefore, in combination with tailored imaging agents, EPRI affords the opportunity to image physiology within the body. In this study, we have prepared water-soluble and membrane-permeant disulfide-linked dinitroxides, at natural isotopic abundance, and with D,(15)N-substitution. Thiols such as glutathione cleave the disulfides, with simple bimolecular kinetics, to yield the monomeric nitroxide species, with distinctive changes in the EPR spectrum. Using the D,(15)N-substituted disulfide-dinitroxide and EPR spectroscopy, we have obtained quantitative estimates of accessible intracellular thiol in cultured human lymphocytes. Our estimates are in good agreement with published measurements. This suggests that in vivo EPRI of thiol-disulfide balance is feasible. Finally, we discuss the constraints on the design of probe molecules that would be useful for in vivo EPRI of thiol redox status.

The cellular architecture of the Daphnia compound eye visual system was studied by using computer-aided techniques. All the neurons in one half of the bilaterally symmetric optic ganglion (OG) were reconstructed in three dimensions from serial electron micrographs. The techniques employed were those developed by Levinthal and collaborators (Macagno, Levinthal, and Sobel, Ann. Rev. Biophys. Bioeng. 8:323-351, 1979). The approximately 200 neurons reconstructed were classified according to where they branch in the OG (the lamina and/or the medulla) and whether they send processes to the supraesophageal ganglion and/or across the midplane. Within each class, neurons were further characterized according to cell body location and size and location of their branching fields. Centrifugal processes from neurons with cell bodies not in the OG were also identified. These results provide the bases for a detailed examination of the synaptic connectivity of the identified neurons and for hypotheses concerning their functional roles in visually evoked behaviors.