Cell death induced by peroxidized low-density lipoprotein: endopepsis.

Peroxidized low-density lipoprotein (p-LDL) has been previously demonstrated to be preferentially cytotoxic to certain malignant cells compared to normal cells of the same type. We present evidence that p-LDL is at least partially taken up through the LDL receptor and that it becomes localized in lysosomes. The integrity of lysosomes of p-LDL-treated cells is compromised, and leakage of their contents into the cytosol occurs.

Effects of tumor necrosis factor-alpha on peroxidation of plasma lipoprotein lipids in experimental animals and patients.

Changes in the plasma lipid composition are observed in patients and animals with malignancy and certain other diseases that are consistent with peroxidation of plasma lipoprotein lipids. These changes can be observed with water-suppressed proton (H-1) and carbon-13 (C-13) nuclear magnetic resonance spectroscopy (NMR) and gas chromatography. Gas chromatography provides evidence of a decrease in polyunsaturated fatty acids relative to monounsaturated fatty acids.

Selective cytotoxicity of low-density lipoprotein to helper T cells of cutaneous T-cell lymphoma after photoperoxidation with 8-methoxypsoralen.

Lymphocyte-containing plasma subjected to photolysis in the presence of 8-methoxypsoralen (methoxsalen, 8-MOP) has previously been shown to be effective against cutaneous T-cell lymphoma and the AIDS-related complex. The mechanism of this effect was thought to involve photoreaction of 8-MOP with DNA, based on certain in vitro experiments. The results of this study suggest a different mechanism.

C-13 NMR spectroscopy of plasma reduces interference of hypertriglyceridemia in the H-1 NMR detection of malignancy. Application in patients with breast lesions.

We have previously described the application of water-suppressed proton nuclear magnetic resonance (H-1 NMR) spectroscopy of plasma for detection of malignancy. Subsequently, hypertriglyceridemia has been identified as a source of false positive results. We now describe a confirmatory, adjunctive technique--analysis of the carbon-13 (C-13) NMR spectrum of plasma--which also identifies the presence of malignancy but is not sensitive to the plasma triglyceride level.

The NMR blood test for cancer: current status.

Our laboratory has developed nuclear magnetic resonance (NMR) techniques for detecting cancer. Using water-suppressed proton (H-1) NMR spectroscopy, we observed that the linewidths of the resonances of methyl and methylene moieties in lipoprotein lipids were consistently narrower in plasma samples from cancer patients than in those from controls. These findings have been corroborated by a number of independent laboratories, but other investigators have been unable to reproduce our results.

Cooperativity in the calcium ion-induced quenching of the intrinsic fluorescence of a series of normal and GLA-deficient bovine prothrombin fragment 1 molecules.

Ca2+ titrations of the intrinsic fluorescence of a series of gamma-carboxyglutamic acid (GLA)-deficient bovine prothrombin fragments 1 yield response Hill plot parameters useful for characterization of the metal ion-binding process. 11-, 10-, and 9-GLA fragments 1 exhibit Tm (the (Ca2+)total concentration at which ln (B/F) = 0 in the response Hill plot) values between 0.2 and 0.

Nuclear magnetic resonance studies of sodium/calcium exchange in frog perfused, beating hearts.

This study explores the effect of extracellular Ca2+ concentration ([Ca2+]o), on the intracellular Na+ concentration ([Na+]i), in frog intact hearts using nuclear magnetic resonance spectroscopy, which allows for the measurement of [Na+]i in perfused, beating hearts. Decreases in [Ca2+]o yielded marked increases in [Na+]i. A similar effect was seen during inhibition of the Na+/K+ pump and was fully reversible.

Measurement of left ventricular mass in rats using electrocardiogram-gated magnetic resonance imaging.

To evaluate the ability of electrocardiogram (ECG)-gated magnetic resonance (MR) imaging to assess in vivo left ventricular (LV) mass in the rat, we studied 20 healthy adult Sprague-Dawley and Fischer 344 rats and 8 additional rats that underwent scanning after induction of volume overload by aortic leaflet disruption. ECG-gated spin-echo pulse sequences were used to acquire a series of 1-mm thick modified short-axis images of the left ventricle. The area enclosed by the endocardial and epicardial borders of the left ventricle was multiplied by the interslice distance and specific gravity of myocardium to obtain calculated slice mass.

Alteration of aliphatic lipid proton NMR linewidths by malignant tumors in guinea pigs.

Water-suppressed proton nuclear magnetic resonance spectroscopy was used to observe plasma lipoprotein lipid methyl and methylene resonances from guinea pigs which had been injected with viable or heat-killed line 1 or line 10 tumor cells or sterile oil. It was shown that the widths of these resonances became significantly sharper as the number of tumor cells grew. Plasma from tumor-free control animals showed no change in the NMR linewidths.

NMR characteristics of "visible" intracellular myocardial potassium in perfused rat hearts.

Nuclear magnetic resonance (NMR) spectroscopy provides a unique view of ions through its noninvasive character and relaxation time measurements. Several previous studies have demonstrated that only approximately 20% of cardiac intracellular potassium is visible with current NMR techniques. This study investigates the NMR visible intracellular potassium in a perfused rat heart preparation.

Cytoplasmic Mg2+ concentration in platelets: implications for determination of Ca2+ with aequorin.

The concentration of cytoplasmic ionized Mg2+ ([Mg2+]i) varies considerably among different cell types. It has not been measured in platelets. Incorrect estimates of this value could markedly affect many intracellular investigations, including calibration of measurements of platelet cytoplasmic ionized Ca2+ concentration ([Ca2+]i) with the photoprotein aequorin and other Ca2+-sensitive probes.

A synthetic functional metabolic compartment. The role of propinquity in a linked pair of immobilized enzymes.

A system was created to model the influence of microcompartments on linked enzymatic reactions. Creatine kinase and hexokinase were covalently attached to Sepharose beads. The gel could be perfused in a specially constructed chamber inside a 360-MHz NMR spectrometer at different flow rates with solutions containing various concentrations of substrates.

Sodium transport and phosphorus metabolism in sodium-loaded yeast: simultaneous observation with sodium-23 and phosphorus-31 NMR spectroscopy in vivo.

Simultaneous 23Na and 31P NMR spectra were obtained from a number of yeast suspensions. Prior to NMR spectroscopy, the yeast cells were Na-loaded: this replaced some of the intracellular K+ with Na+. These cells were also somewhat P-deficient in that they had no polyphosphate species visible in the 31P NMR spectrum.

Nuclear magnetic resonance studies of intracellular ions in perfused frog heart.

Intracellular sodium, potassium, and lithium were observed in a perfused frog heart by nuclear magnetic resonance (NMR) spectroscopy. A perfusate buffer containing the shift reagent, dysprosium tripolyphosphate, was used in combination with mathematical filtering or presaturation of the extracellular resonance to separate the intra- and extracellular sodium NMR signals. Addition of 10 microM ouabain to the perfusate, perfusion with a zero potassium, low-calcium buffer, and replacement of 66% of the perfusate sodium with lithium resulted in the following percent changes in the intracellular sodium levels (mean +/- SD): ouabain + 460 +/- 60% (n = 6), zero potassium + 300 +/- 30% (n = 3), and lithium - 51 +/- 6% (n = 3).

MR imaging of the aortic root and proximal coronary arteries.

Six patients who had recently undergone selective coronary and left ventricular angiography were prospectively examined with MR to show the aortic root and proximal coronary arteries. The examinations were performed with a superconductive 1.5-T instrument with spin-echo sequences and ECG-gated multiple slices of 5-mm thickness.

Intracellular sodium and lithium NMR relaxation times in the perfused frog heart.

We have used a combination of a shift reagent and mathematical filtering or presaturation of the extracellular sodium resonance for the quantitative investigation of the intracellular sodium and lithium relaxation times in the perfused frog heart. While the T1 of the intracellular sodium was found to consist of a single-exponential time constant (approximately 23 ms), the T2 was better fit as a double-exponential decay with time constants of approximately 2 and 17 ms. However, the relative amplitudes of the two time constants in the T2 decay were found to be inconsistent with those which would be expected from a homogeneous pool of nuclei undergoing quadrupolar interactions.

Complete inhibition of creatine kinase in isolated perfused rat hearts.

Transient exposure of an isolated isovolumic perfused rat heart to low concentrations (0.5 mM) of perfusate-born iodoacetamide resulted in complete inhibition of creatine kinase and partial inhibition of glyceraldehyde-3-phosphate dehydrogenase in the heart. At low levels of developed pressure, hearts maintained mechanical function, ATP, and creatine phosphate levels at control values.

Detection of malignant tumors. Water-suppressed proton nuclear magnetic resonance spectroscopy of plasma.

A sensitive and specific blood test for cancer has long been sought. The water-suppressed proton nuclear magnetic resonance (NMR) spectrum of plasma is dominated by the resonances of plasma lipoprotein lipids. We measured the mean line widths of the methyl and methylene resonances, which were found to be correlated with the presence or absence of malignant tumors.

Observation of intracellular potassium and sodium in the heart by NMR: a major fraction of potassium is "invisible".

Using 39K and 23Na NMR in conjunction with extracellularly localized shift reagents, we have determined the intracellular concentrations of NMR visible sodium and potassium in isolated, perfused rat hearts. We find this concentration to be 9.9 mM/kg cell water for sodium and 31 mM/kg cell water for potassium.

Identification and quantitation of O-phosphoserine in human plasma fibronectin.

O-Phosphoserine was positively identified as the phosphorylated moiety in human plasma fibronectin by 31P-NMR of intact peptides. These data correlated completely with chemical analyses which demonstrated the presence of O-phosphoserine at a concentration of 2 residues/molecule. Neither O-phosphothreonine nor O-phosphotyrosine was detected in partial acid and partial alkaline hydrolysates, respectively.