Metabolism of (3-(14)C)tryptophan and (2-(14)C)alanine in cattle tissues

In the experiments involving incubation of the liver, brain cortex, muscle and adipose tissues homogenates with [3-14C] tryptophan for an hour 43.2-89.3% of the label was found in proteins, 7.2-47.2%--in lipids, 2.6-9.4%--in CO2. Following incubation of the above-mentioned tissue homogenates with [2-14C] alanine, proteins, lipids and CO2 contain 28.8-49.3%; 22.6-31.9% and 21.6-49.3% of radioactive label, respectively. Radioactivity of lipids synthesized by the homogenates of the investigated tissues from [3-14C] tryptophan and [2-14C] alanine is 23.5-63.5 and 21.1-56.0%, respectively, the radioactivity of CO2 being 1.4-5.1 and 9.3-11.8% of the above-mentioned compounds synthesized from [1-14C] acetate. The results obtained testify to the considerable contribution of [3-14C] tryptophan and [2-14C] alanine to protein synthesis as well as to their involvement in the substrate supply of lipogenesis and energetic processes in various organs and tissues of cattle.     

Effect of insulin and cortisol on oxidation of (1-14C)glucose, (6-14C)glucose, (1-14C)palmitate and (1-14C)leucine in the tissues of swine during the neonatal period

The intensity of [1-14C]glucose, [6-14C]glucose, [1-14C]palmitate and [1-14C]leucine oxidation and the effect of insulin and hydrocortisone on this process were studied in the brain, duodenum mucosa, liver and skeletal muscle of 1- and 5-day old piglets in vitro. Most of the studied substrates are oxidized in the tissues of 5-day piglets more intensively. Insulin stimulates oxidation of [1-14C]glucose, [6-14C]glucose and [1-14C]leucine in the brain and duodenum mucosa in 1- and 5-day old piglets, while in the liver and skeletal muscle--only in 5-day old piglets. Hydrocortisone administration enhances oxidation of [1-14C]leucine in most of the studied tissues in 1-day piglets and oxidation of [1-14C]glucose and [6-14C]glucose--in 5-day piglets. Both hormones produce no essential influence on the intensity of [1-14C]palmitate oxidation in the studied tissues of piglets or somewhat weaken it.     

Preparation and Metabolism of 2-(14C)-cis, trans-Xanthoxin

2-[¹⁴C]-cis, trans-xanthoxin (specific activity 0.26 µCi/µmol)has been synthesized from4-(1', 2'-epoxy-4'-hydroxy-2', 6',6'-trimethyl-l'-cyclohexyl)-trans-3-buten-2-oneand methyl 2-[¹⁴C]-bromoacetate. When 2-[¹⁴C]-cis, trans-xanthoxin was fed to cut shoots of tomatoand dwarf bean, it was converted within 8 h to (+)-abscisicacid in yields of 10.8 and 7.0 per cent respectively. Pea seedsand tomato fruits gave much smaller conversions. A second majormetabolic product extracted from treated tomato and bean shootswas shown to be a metabolite of abscisic acid and has been tentativelyidentified as phaseic acid. These results, together with the earlier finding that xanthoxinis present in the extracts of many seedlings, suggest that xanthophyllsand xanthoxin may be precursors in the biosynthesis of ( + )-abscisicacid     

Conversion of (U-14C)-glycerol, (2-3H)-glycerol and (1-14C)-palmitate into circulating lipoproteins in the rat.

The in vivo formation of labelled very low density lipoproteins (VLDL) from (U-14C)-glycerol, (2-3H)-glycerol and (1-14C)-palmitate was studied in fed female rats. The rate of disappearance of radioactivity from plasma after the i.v. injection with these tracers was similar for (U-14C)-glycerol and (1-14C)-palmitate. With (2-3H)-glycerol, plasma radioactivity at 10 min was lower than with the other substrates although it did not change thereafter. A certain proportion of radioactivity administered as glycerol appeared in plasma lipids, mainly in the VLDL glyceride glycerol fraction, although when (U-14C)-glycerol was the substrate, a considerable portion also appeared in the esterified fatty acids of these lipoproteins. When using (1-14C)-palmitate, practically all the circulating labelled esterified fatty acids appeared in the VLDL fraction, while the labelled free fatty acids appeared in lipoprotein of higher density, presumable free fatty acid-albumin complexes. This data is discussed in terms of the role of the liver in the rapid, continuous cycling of these substrates to yield VLDL-glycerides for their extrahepatic utilization.     

Lipogenesis from glucose-2- 14 C and acetate-1- 14 C in aorta

Lipogenesis was measured with glucose-2-(14)C and acetate-1-(14)C in the everted aortas of normal and atherosclerotic rabbits. More glucose-2-(14)C than acetate-1-(14)C was incorporated into lipids in both the normal and the atherosclerotic aorta. Radiocarbon from glucose-2-(14)C appeared mainly in triglycerides and phospholipids with a small amount in cholesteryl esters. Incorporation increased almost threefold with atherosclerosis, most of the radioactivity being in the glycerol moiety; radioactivity was predominantly in carbon 2 of glycerol. About 70% of the acetate-1-(14)C incorporated into phospholipids and triglycerides was in the fatty acids, and the remainder was in glyceride-glycerol; 98% of the radioactivity in cholesteryl esters was in the fatty acid moiety. Incorporation into cholesteryl esters was increased most during the development of atherosclerosis. Fatty acid synthesis was similar from both acetate-1-(14)C and the 2 carbon unit derived from glucose-2-(14)C, viz., predominantly de novo synthesis of fatty acids with 14 and 16 carbon atoms, and elongation for those of 18 carbons and longer.     

The metabolic fate of (2-14C)folic acid and a mixture of (2-14C)- and (3'',5'',9-3h)-folic acid in the rat.

The metabolism of [2-14C]folic acid over 13 days and a mixture of [2-14C]- and [3',5',9-3h]-folic acid in rats over a 6-day period is described. Both 14C and 3H are excreted in urine over the 6-day period, but 3H and 14C are only detectable in faeces for 2 days. A breakdown product of folic acid labelled with 3H only was found in some urine samples, but no metabolite corresponding to the part of the molecule containing 14C was detected. These experiments show that in the whole animal a substantial portion of orally administered folic acid undergoes scission shortly after administration [Blair Biochem. J. (1957) 68, 385-387] and that the retained folates are a shortage form for folate monoglutamates.     

Metabolism of 2-hydroxy(4-14C)estradiol by rat liver microsomes

The metabolism of ¹⁴C-labeled estradiol and 2-hydroxyestradiol by rat liver microsomes has been compared under a variety of experimental conditions. An active liver microsomal system as well as NADPH was required to obtain high percentage yields of water-soluble metabolites from both steroids. Spermine stimulated the formation of polar products in both cases but the sex-difference in metabolism observed with estradiol was less marked with 2-hydroxyestradiol as substrate.     

Hydrolysis of glyceryl tri(1- 14 C)octanoate and glyceryl tri(1- 14 C)oleate monolayers by postheparin lipolytic activity

The hydrolysis of monomolecular films of glyceryl tri[1-(14)C]octanoate and glyceryl tri[1-(14)C]oleate has been demonstrated by measurement of the decrease in surface radio-activity that occurs in the presence of postheparin plasma. The hydrolysis displayed first order kinetics and was proportional to enzyme concentration over a 10-fold range. No hydrolysis was observed in the absence of enzyme, and only slight activity (1%) was found in plasma taken from subjects before heparin administration. The hydrolysis was stimulated to a variable extent by Ca(2+). The first product of hydrolysis of the monolayer was identified as 1,2-diglyceride, which was subsequently converted to 2-monoglyceride. Inhibition of triglyceride hydrolysis was observed when postheparin plasma was preincubated in 2 m NaCl, 10(-4) m protamine, 10 mm Na(4)P(2)O(7), and 0.1 m NaF. Monolayer techniques avoid some but not all of the problems associated with emulsified lipid substrates and appear to be applicable for study of post-heparin lipolytic activities.     

The Long Term Metabolism of ({+/-})-(2-14C)Abscisic Acid by Apple Seeds

Apple seeds soaked in a solution of [±]-[2-¹⁴C]abscisicacid for up to 70 d formed phaseate and the epimeric dihydrophaseatesand all four acids were converted into alkali-hydrolysable conjugates.This metabolism occurred in the husks and in the embryos. Bothparts of untreated control seeds contained the four acids andtheir conjugates. In addition to these characterized metabolites,a number of other, labelled, acidic products were isolated fromthe seeds. ¹⁴CO₂ was evolved from unsterilized seeds but notfrom surface-sterilized ones. Bacterial cultures isolated fromsoil and rotting fruit metabolized [¹⁴C]ABA to a range of compounds,but not to phaseic or the dihydro-phaseic acids. A new, acidicconjugate of ABA is described.