The 31P Spectral Modulus (PSM) as an Assay of Metabolic Status.

Abstract

The phosphorus-31 (³¹P) spectral modulus (PSM) is a measure of the metabolic status of cells, tissues, and organs. The PSM can be calculated from ³¹P nuclear magnetic resonance (³¹P NMR) spectra obtained from cell, tissue, or organ preparations. These ³¹P NMR spectra can be a measure of intact living cells, tissues, or organs, or appropriate biochemical extracts of such preparations. The ³¹P NMR spectrum is comprised of signals derived from organophosphate metabolites that resonate from 10 δ to -25 δ on the phosphorus chemical shift δ scale. The PSM is the ratio of the high-energy phosphate to that of the low-energy phosphate spectral integrals. These integrals may be conveniently grouped into high-energy and low-energy spectral regions, respectively, into ³¹P chemical shifts located between -0.13 δ to -25 δ and between 10 δ to -0.13 δ. High-energy phosphates are typically described as providing the energy necessary for the activity of cellular metabolism; chemically, they contain one or more phosphate anhydride bonds. This study demonstrates that, (1) in general, the higher the metabolic activity, the higher the PSM, and (2) the modulus calculation does not require a highly resolved ³¹P spectrum and can be calculated solely from the integral. The PSM was calculated among cells, tissues, and organs considered normal, diseased, and stressed. In diseased (mean 1.29 ± 0.73) and stressed (mean 1.23 ± 0.75) cells, tissues, and organs, PSM values are typically low or low relative to normal cells, tissues, or organs (mean 1.65 ± 0.90), following time-course measurements, in dynamic decline. The PSM is useful in determining the metabolic status of cells, tissues, or organs and can be employed as a calculable numeric assay for determining health status statically or over time. Calculation of the PSM can be carried out with spectra of low signal-to-noise; it relies on the minimal resolution required to detect an integral curve having a clear spectral integral inflection point at ca. -0.13 δ. Detection of an integral curve alone enables the calculation of a PSM even at levels of phosphorus concentration so low as to prevent detection of the individual or groups of metabolites, such as with in vivo or ex vivo cell, tissue, or organ determinations. This study (1) presents the foundations and fundamentals of the PSM, a living index of tissue metabolic health, and (2) demonstrates the use of spectral scan analysis in opening new vistas of biology and medicine for measuring the metabolic status of stressed and diseased tissues at a range of detectable levels for monitoring therapeutic interventions.