Identification by nuclear magnetic resonance and mass spectrometry of a glucuronic acid conjugate of o-hydroxybenzoic acid in normal urine and uremic plasma.

Abstract An endogenous compound (included in the fraction of uremic toxins often called the "uremic middle molecules") was separated from plasma of uremic patients and urine from normal persons. As elucidated by mass spectrometry, enzymatic hydrolysis, and 1H, 13C nuclear magnetic resonance, it appears to be a conjugate of o-hydroxybenzoic acid with glucuronic acid. Its presence in urine of healthy subjects indicates its physiological character.</jats:p

23Na nuclear magnetic resonance study of Na+-K+ pump inhibition by a fraction from uremic toxins.

Abstract An in vitro inhibitor of Na+/K+-transporting ATPase (EC 3.6.1.37) was isolated from uremic plasma and normal urine by liquid chromatography. A 23Na nuclear magnetic resonance study involving living erythrocytes showed that this inhibitor causes impairment of the Na+-K+ pump of intact erythrocytes. This finding may explain the high intra-erythrocytic sodium concentration in those uremic patients exhibiting a high concentration of this inhibitor. The presence of this same inhibitor in normal urine suggests that it may play a physiological role.</jats:p

A compound from uremic plasma and from normal urine isolated by liquid chromatography and identified by nuclear magnetic resonance.

Abstract A compound present in normal urine and in ultrafiltrates of uremic plasma in the fraction of so-called "uremic middle molecules" was isolated by liquid chromatography. Preliminary studies, including amino acid analysis, characterization of uronic acids, and ultraviolet spectroscopy, show that the molecule contains glycine, a uronic acid, and an aromatic ring. Characterization by 1H and 13C nuclear magnetic resonance spectrometry shows conclusively that this compound is a double conjugate of glucuronidate--o-hydroxyhippuric acid, which has been previously described by Zimmerman et al., using quite different techniques of isolation and identification (Clin Nephrol 14: 107, 1980; FEBS Lett 129: 237, 1981).</jats:p

Intraerythrocytic pH variations during hemodialysis: A 31P NMR study

Intraerythrocytic pH variations during hemodialysis: A 31P NMR study. Before hemodialysis, patients have an intraerythrocytic pH (pHi) and an extracellular pH, measured in whole blood (pHo), which are lower than those of healthy controls. During bicarbonate hemodialysis, pHi values continuously increase, approaching a normal value at the end of the session. Concomitantly, pHo values follow similar variations. During acetate hemodialysis, pHi values exhibit a steep initial decrease, reaching a minimum after about 15 minutes. Concurrently, however, pHo values decrease only slightly. This phenomenon seems to originate in the intraerythrocytic medium and might be due to a shift in intracellular CO2/bicarbonate equilibrium. This drop in pHi exhibits interpatient variability, suggesting that the magnitude of pH decrease would be correlated with the degree of the problems observed in some patients undergoing acetate hemodialysis

Modification of intra-erythrocytic homeostasis in uremic patients, as studied with 31P nuclear magnetic resonance.

Abstract Intra-erythrocytic pH, ATP concentrations, and 2,3-diphosphoglycerate relaxation times were studied in living erythrocytes by "high-resolution" 31P NMR spectroscopy to assess homeostasis within the cells. In uremic patients, intra-erythrocytic pH is significantly decreased before hemodialysis, but is corrected equally well by hemodialysis against either acetate or bicarbonate. This acidic pHi may be correlated with the increased concentration of ATP in erythrocytes in uremia, which is partly corrected by these two types of hemodialysis. Similarly, the significant decrease of spin-spin relaxation times in uremic patients is corrected by hemodialysis.</jats:p

Kinetic Modeling of Intracellular pH and Comparison with ³¹P NMR Experimental Values in Dialysed Uremic Patients

Changes in intra-erythrocytic pH values over time, during and after bicarbonate hemodialysis, were studied with 31P Nuclear Magnetic Resonance. Simultaneously, pH values of whole blood were obtained by a gazometric method. A two-compartment model appeared to be the simplest kinetic model to explain the shifts in proton concentrations in extra- and intracellular media. Non-linear regression was used to determine exchange constant values. There was a very good correlation between the experimental and calculated proton concentrations. This model can describe all patients but individual experimental constants must be determined. Under these conditions a single blood pH determination before dialysis will permit determination of the initial intra-erythrocytic pH and monitoring of intra-erythrocytic pH during hemodialysis. </jats:p

Intra-erythrocytic sodium in uremic patients, as determined by "high-resolution" 23Na nuclear magnetic resonance.

Abstract The use of 23Na nuclear magnetic resonance with aqueous shift reagent has made it possible to determine intracellular sodium concentrations in living erythrocytes. We applied this technique to samples from 16 healthy subjects and 41 uremic patients. The results seem to show distinct populations among the latter. Classically, two different relaxation times are obtained for intracellular sodium in biological media, according to relaxation NMR theory. Some patients, however, exhibit abnormal results that cannot be accounted for by this theory.</jats:p