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Alpha-aminoisobutyric acid efflux from the cornea of the toad, Bufo marinus.

1. Amino acids move into and out of the amphibian cornea across its inner aqueous, side only. Most alpha-aminoisobutyric acid (AIB) accumulation takes place in the corneal epithelium; the endothelium and stroma do not limit exchanges with these cells. The apical, or tear, surface of the epithelium is an impermeable barrier to the transport of amino acids. There are several sites or mechanisms by which AIB enters and leaves the cornea. 2. The entry of AIB is not Na-dependent; however, the exit site is very sensitive to changes in internal Na concentration. Any factor, such as ouabain or metabolic inhibitors, that increases internal Na, markedly stimulates AIB efflux. 3. Site are also present for the exchange of internal for external amino acids, and this process is Na-dependent. There was no measurable movement of Na into or out of the cells with these amino acids. Exchange efflux was more specific than uptake, since both alanine and leucine inhibit AIB uptake, but only alanine stimulates AIB efflux. 4. Although the largest amount of AIB accumulated by the cornea was present in the epithelium, evidence is presented that the endothelium and stromal keratocytes may also concentrate and retain amino acids.

Aminoisobutyric Acids↗

Roles of Na+ and K+ in alpha-aminoisobutyric acid transport by the marine bacterium Vibrio alginolyticus.

Effects of monovalent cations on alpha-aminoisobutyric acid (AIB) transport were examined in the marine bacterium Vibrio alginolyticus. In K+-containing cells, AIB was actively accumulated only in the presence of Na+, and the addition of K+ had essentially no effect. On the other hand, K+-depleted and Na+-loaded cells required K+ as well as Na+ for the accumulation of AIB against its concentration gradient. The characterization of the roles of Na+ and K+ in AIB transport was performed by manipulation of intra- and extracellular cation compositions. K+ concentration gradient (K+in greater than K+out) was not essential for the Na+-dependent AIB uptake. Na+ extrusion against its concentration gradient in Na+-loaded cells occurred only in the presence of K+(Rb+). Half-maximal stimulations of the Na+ extrusion and AIB uptake by K+ were observed at K+ concentration near apparent Km for K+ transport. Finally, in the presence of the Na+ electrochemical gradient (toward the inside), K+ was not necessary for AIB uptake. From these results, it was concluded that the Na+-dependent AIB uptake is driven by the Na+ electrochemical gradient across the membrane and that K+ is required for AIB uptake only for the generation of the Na+ electrochemical gradient.

Aminoisobutyric Acids↗

Rapid determination of beta-aminoisobutyric acid by reversed-phase high-performance liquid chromatography.

For the determination of beta-aminoisobutyric acid (BAIBA) in urine samples in which the beta-alanine concentrations are higher than those of BAIBA, the resolution between these two amino acids, separated by reversed-phase liquid chromatography on an octadecylsilane column, was optimized. The chromatographic analysis included precolumn derivatization of amino acids with o-phthalaldehyde, followed by a 15-min isocratic elution and detection at 340 nm. Because of its simplicity, this method should be useful for monitoring urinary excretion of BAIBA.

Aminoisobutyric Acids↗

Denervation stimulation of alpha-aminoisobutyric acid uptake by the diaphragm in developing rats.

Effect of denervation on accumulation of alpha-aminoisobutyric acid (AIB) by the the diaphragm was studied in 11-, 21-, 31-, and 51-day-old rats. In both innervated and denervated diphragms the accumulation steadily decreased with age. Denervation significantly enhanced accumulation at all ages, the relative increment being greater in older animals owing to the lower basal accumulation ratio for AIB. The denervation-induced stimulation was partially abolished by dactinomycin administration. The drug obviously inhibits the formation of new carrier sites at plasma membranes for AIB transport.

Age Factors↗

Tumor imaging with carbon-11 labeled alpha-aminoisobutyric acid (AIB) in patients with malignant melanoma.

The potential usefulness of [C-11]-labeled alpha-aminoisobutyric acid (AIB) for tumor imaging has been demonstrated previously in our findings of increased tumor uptake with C-14-labeled AIB in human melanoma heterotransplants in nude mice, and subsequently, in a single case study using C-11 AIB to demonstrate the extent of metastases in patient with widespread malignant melanoma. We report here on the use of C-11 AIB in ten patients with metastatic or unresectable malignant melanoma. Five patients had intense tracer uptake at all known sites of tumor involvement. A sixth patient had good uptake in metastatic lesions in the shoulder and the pelvis, but did not demonstrate uptake within metastatic lesions in the lungs. Two patients had only minimal uptake over the tumor lesions while the two other patients had essentially normal studies. Further studies with C-11 AIB in patients with melanoma and other tumors are warranted.

Adult↗

Alpha-aminoisobutyric acid loss by rat small intestine: role of intestinal site and luminal contents.

Loss of amino acid into the lumen may be important in diarrheal states. Labeled alpha-aminoisobutyric acid (AIB) was given parenterally, and concentration gradients developed between lumen, intestinal tissue and plasma. After 24 h, we measured AIB loss into the intestinal lumen by recirculating isolated segments in situ under conditions of either net water absorption or net water secretion. Final 14C-AIB concentrations in intestinal tissue were 200-800 times that of lumen, 6--15 times that of serum. Despite increased total 14C-AIB loss during net water secretion, and based on the low luminal 14C-AIB concentrations, the contribution of amino acid loss to fecal nitrogen during osmotic diarrhea appears to be minimal.

Aminoisobutyric Acids↗

Aminoisobutyric acid transport in soleus muscles of lean and gold thioglucose-obese mice.

The uptake of alpha-aminoisobutyric acid (AIB) was studied in soleus muscles isolated from lean and gold thioglucose-lesioned obese mice (GTG-obese), both in basal and in insulin-stimulated conditions. The basal (i.e., non-insulin-stimulated) influx of AIB was decreased in muscles of GTG-obese mice because of a decrease in the apparent Vmax of the transport system. In muscles from both types of mice, insulin stimulated AIB influx by increasing the apparent Vmax of transport; this effect was partially dependent on new protein synthesis. In muscles of GTG-obese mice, the effect of insulin was altered in two ways: the absolute value of the maximally insulin-stimulated AIB influx was lower than in muscles of lean controls, and the dose response of insulin-stimulated AIB influx was shifted to the right (half-maximally effective concentration, EC50 congruent to 1.7 nM) compared to that observed in muscles of lean animals (EC50 congruent to 0.7 nM). It is concluded that in muscles of obese mice a) amino acid transport per se is altered, and b) the sensitivity of this process to the stimulatory effect of insulin is markedly diminished.

Aminoisobutyric Acids↗

Characteristics and energy requirements of an alpha-aminoisobutyric acid transport system in Streptococcus lactis.

Galactose-grown cells of Streptococcus lactis ML3 acculated alpha-aminoisobutyric acid (AIB) by using energy derived from glycolysis and arginine catabolism. The transport system displayed low-affinity Michaelis-Menten saturation kinetics. Using galactose or arginine as energy sources, similar V max and K m values for AIB entry were obtained, but on prolonged incubation the intracellular steady-state concentration of AIB in cells metabolizing arginine was only 65 to 70% that attained by glycolyzing cells. Efflux of AIB FROM PRELOADED CElls was temperature dependent and exhibited the characteristics of a first-order reaction. The rate of AIB exit was accelerated two- to threefold in the presence of metabolizable energy sources. Metabolic inhibitors including p-chloromercuribenzoate, dinitrophenol, azide, arsentate, and N, N'-dicyclohexylcarbodiimide either prevented or greatly reduced AIB uptake. Fluoride, iodoacetate and N-ethylmaleimide abolished galactose-dependent, but not arginine-energized, AIB uptake. K+ and Rb+ reduced the steady-state intracellular AIB concentration by approximately 40%, and these cations also induced rapid efflux of solute from actively transporting cells. Equivalent concentrations (10 mM) of Na+, Li+, or NH4+ were much less inhibitory. The proton-conducting ionophores tetrachlorosalicylanilide and carbonylcyanide m-chlorophenlyhydrazone abolished uptake and induced AIB efflux even though glycolysis and arginine catabolism continued at 60 and 140%, respectively, of control rates. A proton motive force is most likely involved in the active transport of AIB, whereas data from efflux studies suggest that energy is coupled to AIB exit in cells of S. lactis ML3.

Aminoisobutyric Acids↗

Transport of alpha-aminoisobutyric acid by Streptococcus pyogenes and its derived L-form.

We studied the uptake of alpha-aminoisobutyric acid (AIB) in Streptococcus pyogenes and its physiologically isotonic L-form. S. pyogenes cells starved for glucose or treated with carbonyl cyanide-m-chlorophenyl hydrazone accumulated limited amounts of AIB. A high apparent K(m) value characterized the glucose-independent transport of AIB. The rate and extent of AIB accumulation significantly increased in the presence of glucose. Two saturable transport components with distinct apparent K(m) values characterized glycolysis-coupled transport of AIB. A biphasic Lineweaver-Burk plot was also obtained for l-alanine transport by glycolyzing S. pyogenes cells. AIB seems to share a common transport system(s) with glycine, l- and d-alanine, l-serine, and l-valine. This was shown by the competitive inhibition of AIB uptake by these compounds and their ability to induce competitive exchange efflux of accumulated AIB. About 30% of the AIB uptake was not inhibited by a saturating amount of l-valine, indicating the existence of more than one system for AIB transport. p-Chloromercuribenzoate markedly inhibited the accumulation of AIB by both glycolyzing and glucose-starved cells. In contrast, carbonyl cyanide-m-chlorophenyl hydrazone affected only metabolism-dependent uptake of AIB, which was also sensitive to dinitrophenol, N-ethylmaleimide, iodoacetate, fluoride (NaF), arsenate, and N,N'-dicyclohexylcarbodiimide. These results are interpreted according to the chemiosmotic theory of Mitchell, whereby a proton motive force constitutes the driving force for AIB accumulation. AIB was not accumulated by the L-form. However, a temporary accumulation of AIB by a counterflow mechanism and a saturable system with a low apparent affinity were demonstrated for AIB transport by this organism. We suggest that a deficiency in the coupling of energy to AIB transport is responsible for the apparent lack of active AIB accumulation by the L-form.

Alanine↗

Relation between urinary beta-aminoisobutyric acid excretion and concentration of lead in the blood of workers occupationally exposed to lead.

Urinary beta-aminoisobutyric acid (ABA) concentration was determined by fluorometric high performance liquid chromatography in 22 workers occupationally exposed to lead. The urinary excretion of ABA was increased with increasing exposure to lead. The excretion of urinary ABA had a significant correlation with concentration of lead in blood (Pb-B) (r = 0.581), similar to the correlation of Pb-B with urinary delta-aminolaevulinic acid (ALA) concentration (r = 0.563). The determination of urinary ABA concentration in workers exposed to lead, therefore, may offer a new approach for evaluating the health effect of lead.

Adult↗

The accumulation of 2-aminoisobutyric acid in the costal and lumbar parts of the phrenectomized rat diaphragm in vitro.

The accumulation of 2-aminoisobutyric acid initially increased in denervated costal and lumbar parts of the rat hemidiaphragm after both unilateral and bilateral phrenectomy, followed by a later gradual decrease. The enhancement of accumulation was invariably higher in the costal than in the lumbar part. The accumulation was initially lower in bilaterally than in unilaterally phrenectomized hemidiaphragms, but the relationship was reversed after 72 h due to a faster reattenuation of the enhancements in unilaterally phrenectomized diaphragms.

Aminoisobutyric Acids↗

Regulation of polyamine synthesis by dietary alpha-aminoisobutyric acid and ornithine.

An experiment was conducted to determine the effect of feeding ornithine in combination with alpha-aminoisobutyric acid (AIB), an inhibitor of arginase, on the regulation of polyamine synthesis in chicks. A total of 48 chicks with genetically elevated renal arginase activity was fed diets containing crystalline amino acids and 1% AIB with or without 2% ornithine. Feeding AIB reduced renal arginase activity, while renal and hepatic ornithine decarboxylase (ODC) activity increased. Feeding AIB plus ornithine caused no further reduction in renal arginase activity compared with that in chicks fed the AIB-supplemented diet. Renal and hepatic ODC activities, however, fell to below control levels. Renal, hepatic, and breast muscle ornithine concentrations increased substantially when ornithine was fed. AIB plus ornithine increased renal putrescine and spermidine concentrations. It was concluded that AIB could partially overcome the ornithine-induced inhibition of ODC activity. These findings support the hypothesis that dietary manipulation of precursor amino acids of polyamines in the presence of metabolites that induce ODC activity can influence tissue polyamine concentrations.

Adenosylmethionine Decarboxylase↗

Effects of maternal ethanol ingestion on the uptake of alpha-aminoisobutyric acid by fetal rat liver, lung and brain.

Maternal ethanol ingestion reduced the uptake of [14C]-alpha-aminoisobutyric acid (AIB) by placenta and fetal liver, lung and brain when compared with pair-fed or ad libitum-fed controls. The ratios between fetal tissue and plasma radioactivities indicated reduced AIB transport into the tissues of the ethanol-fed fetuses. Organ weights correlated with the fetal:maternal plasma 14C ratio, an indicator of the rate of placental AIB transfer, and with the tissue AIB uptake. These observations suggest that ethanol-induced growth retardation may reflect reduced nutrient availability resulting from impaired transport processes.

Aminoisobutyric Acids↗

Increased urinary excretion of beta-aminoisobutyric acid in a Danish family.

During collection of a control material for determination of urinary excretion of beta-aminoisobutyric acid (beta-AIB), female high excretors were found in four generations in a Danish family. The heredity seemed to be sex related and dominant, unlike previous communications about genetically conditioned high excretors of beta-AIB.

Adolescent↗

The effect of endotoxin on plasma alpha-aminoisobutyric acid.

The i.v. injection of bacterial endotoxin into dogs was found to cause a rapid increase in plasma levels of infused alpha-aminoisobutyric acid. The findings suggest that nonmetabolic factors (tissue uptake, fluid shifts) influence amino acid distribution during endotoxemia.

Aminoisobutyric Acids↗

Studies of the uptake and distribution of inulin and alpha-aminoisobutyric acid after intraamniotic injection in the guinea pig.

Pregnant guinea pigs were given intraamniotic injection of 14C-alpha-aminoisobutyric acid and 3H-inulin. Both substances were taken up by the fetus in different concentrations. High levels of alpha-amino-isobutyric acid were found in the placenta, liver, and serum. Inulin on the other hand, being a macro-molecule distributed by diffusion in the extracellular space, was found in high concentrations in the small intestine and in the placenta. Thus the main pathway for transferring nutrient substances from the amniotic fluid to the fetus would seem to be the gastro-intestinal tract. Thus supplementary nutrition of the fetus by the intraamniotic injection of nutrients is a possibility.

Aminoisobutyric Acids↗

Solid-phase separation of 14C-labelled urea and aminoisobutyric acid for membrane transport studies.

A sorbent extraction method has been developed for separating 14C-labelled urea and aminoisobutyric acid (AIB) in blood. The use of commercial solid-phase extraction cartridges containing aminopropyl-bonded silica provided a convenient and rapid separation of urea and AIB with better than 92% recovery of each and less than 5% cross-contamination. This allows these compounds, together with [3H]methylglucose, to be used as marker compounds for investigating three aspects of membrane transport. The facility to separate any two of the three compounds permits their simultaneous measurement, greatly increasing the amount of data obtainable from each in vivo preparation.

3-O-Methylglucose↗