Katherine Liu

13C NMR study of hepatic pyruvate carboxylase activity in tumor rats

Alanine and lactate, as major gluconeogenic substrates, must be converted into oxaloacetate by way of pyruvate carboxylase before their entry into gluconeogenesis. Although it is well known that hepatic gluconeogenesis from these substrates is increased in tumor hosts, the involvement of pyruvate carboxylase has not been demonstrated. In the present study, we examined pyruvate carboxylase activity in the perfused livers of tumor rats using 13C NMR spectroscopy with [3-13C]-alanine as the gluconeogenic substrate. A substantial increase in hepatic [3-13C]-aspartate production was found in the tumor rats. Since aspartate accumulation directly reflects fluxes of alanine through pyruvate carboxylase, the observed increase in hepatic production of [3-13C]-aspartate in tumor rats indicates that pyruvate carboxylase activity is significantly enhanced.

The nature and subtlety of abnormal findings in chest radiographs.

All detected abnormal findings were recorded for 1085 consecutive chest x-ray examinations. Each finding was classified by descriptive criteria and graded for subtlety. Accuracy of interpretation was evaluated in a randomized sample of 100 cases using follow-up or multiple readers. Seventy percent of standard examinations and 93% of bedside examinations revealed abnormal findings. Pulmonary infiltrates were the most commonly detected abnormality, being present in 55% of abnormal cases. Noncalcified pulmonary nodules and pneumothoraces were each present in approximately 5% of abnormal cases. The most commonly encountered subtle findings were due to intravenous catheters, pulmonary infiltrates, pneumothoraces, rib lesions, and pulmonary nodules in descending order of frequency. It is concluded that it is reasonable to use selected examples of these findings in observer tests when evaluating new imaging modalities such as digital radiography.

Gluconeogenesis in the liver of tumor rats.

Altered gluconeogenesis is frequently observed in cancerous hosts. To define its derangements in the liver, we studied glucose and glycogen production in the perfused livers of tumor-bearing rats using 13C NMR spectroscopy. Nine Fischer 344 rats were inoculated with mammary adenocarcinoma. After 5 weeks, the livers were removed and perfused with Krebs buffer containing 8 mM L-[3-13C]alanine, and 13C NMR spectroscopy was performed. Nine pair-fed rats were studied as controls. The peak heights of glucose and glycogen in the 13C NMR spectra of the perfused livers and final perfusates of the two groups of rats were compared. We found comparable amounts of C1-labeled glucose and glycogen in the two groups, but C2- to C5-labeled and C6-labeled glucose and glycogen, as well as total 13C-labeled glucose and glycogen, appeared in smaller quantities in the tumor rats than in the pair-fed rats. These findings suggest that appreciable amounts of unlabeled glycerol were utilized by both groups, but less so by the tumor rats than the pair-fed rats. In addition, there was decreased production of oxaloacetate through pyruvate dehydrogenase and the Krebs cycle in the livers of the tumor rats, where the overall metabolism of alanine into glucose and glycogen was also reduced.

Gluconeogenesis in the Livers of Diet-Restricted Rats-A 13C Nuclear Magnetic Resonance Study

BACKGROUND Gluconeogenic activity is reduced during starvation. However, it is less clear whether the utilization of gluconeogenic substrates is diminished with mild but prolonged diet restriction and, if so, whether there are intrinsic changes in the gluconeogenic pathway. We examined gluconeogenesis in the livers of diet-restricted rats with 13C nuclear magnetic resonance (NMR) spectroscopy. METHODS Fischer 344 rats were given 88% (DR group) of what was consumed by the weight-matched ad libitum-fed normal rats (CL group). At the end of 5 weeks, the removed livers were perfused with [3-13C] alanine while 13C NMR spectroscopy was performed. RESULTS The final body and liver weights were the same for the two groups. In DR rats, both intrahepatic [3-13C] alanine and metabolites generated via pyruvate and oxaloacetate, including aspartate and carbamoyl aspartate, appeared in significantly reduced amounts. There was also marked diminution in the production of glucose. CONCLUSIONS In the livers of DR rats, alanine uptake through System A transport, the fluxes through pyruvate carboxylase, the biosynthesis of pyrimidine nucleotides, and the production of glucose from alanine were all significantly decreased with mild intake restriction. Attenuated protein synthesis in the liver of diet-restricted animals may be the cause for this decreased utilization of alanine for gluconeogenesis.