Comparison of Arterial-Venous Balance and Tracer Incorporation Methods for Measuring Muscle Fractional Synthesis and Fractional Breakdown Rates.

Loss of muscle mass in response to injury or immobilization impairs functional capacity and metabolic health, thus hindering rehabilitation. Stable isotope techniques are powerful in determining skeletal muscle protein fluxes. Traditional tracer incorporation methods to measure muscle protein synthesis and breakdown are cumbersome and invasive to perform in vulnerable populations such as children. To circumvent these issues, a two-bolus stable isotope amino acid method has been developed; although, measured rates of protein synthesis and breakdown have not been validated simultaneously against an accepted technique such as the arterial-venous balance method. The purpose of the current analysis was to provide preliminary data from the simultaneous determination of the arteriovenous balance and two-bolus tracer incorporation methods on muscle fractional synthesis and breakdown rates in children with burns. Five were administered a primed-constant infusion of L-[15N]Threonine for 180 minutes (Prime: 8 µmol/kg; constant: 0.1 µmol·kg-1·minute-1). At 120 and 150 minutes, bolus injections of L-[ring-13C6]Phenylalanine and L-[15N]Phenylalanine (50 µmol/kg each) were administered, respectively. Blood and muscle tissue samples were collected to assess mixed muscle protein synthesis and breakdown rates. The preliminary results from this study indicate that there is no difference in either fractional synthesis rate (mean ± SD; arteriovenous balance: 0.19 ± 0.17 %/h; tracer incorporation: 0.14 ± 0.08 %/h; P = .42) or fractional breakdown rate (arteriovenous balance: 0.29 ± 0.22 %/h; tracer incorporation: 0.23 ± 0.14 %/h; P = .84) between methods. These data support the validity of both methods in quantifying muscle amino acid kinetics; however, the results are limited and adequately powered research is still required.

Translating slow-binding inhibition kinetics into cellular and in vivo effects.

Many drug candidates fail in clinical trials owing to a lack of efficacy from limited target engagement or an insufficient therapeutic index. Minimizing off-target effects while retaining the desired pharmacodynamic (PD) response can be achieved by reduced exposure for drugs that display kinetic selectivity in which the drug-target complex has a longer half-life than off-target-drug complexes. However, though slow-binding inhibition kinetics are a key feature of many marketed drugs, prospective tools that integrate drug-target residence time into predictions of drug efficacy are lacking, hindering the integration of drug-target kinetics into the drug discovery cascade. Here we describe a mechanistic PD model that includes drug-target kinetic parameters, including the on- and off-rates for the formation and breakdown of the drug-target complex. We demonstrate the utility of this model by using it to predict dose response curves for inhibitors of the LpxC enzyme from Pseudomonas aeruginosa in an animal model of infection.

Threonine peptidases as drug targets against malaria.

INTRODUCTION: Malaria is caused by the intracellular parasite Plasmodium falciparum. Although numerous therapies are available to fight the disease, the number of pharmacophores is small, and constant development of novel therapies, especially with new targets, is desirable to fight developing resistance against presently prescribed drugs. AREAS COVERED: This review discusses research on plasmodial threonine peptidases along with recent advances in proteasome inhibitor development. EXPERT OPINION: While PfHslV is an attractive drug target in malaria, more investigation is required to clarify its functional role in the parasite. More efforts should also be invested in assessing the plasmodial proteasome as a drug target. The few papers investigating the effect of proteasome inhibitors on different stages of the life cycle point towards important roles not only during asexual, but also in hepatic and sexual stages, in humans and the mosquito. If this holds true, this is a key argument to further develop proteasome inhibitors for use against malaria.

Development and validation of a high-performance liquid chromatography-tandem mass spectrometry method for the determination of the novel proteasome inhibitor CEP-18770 in human plasma and its application in a clinical pharmacokinetic study.

CEP-18770, [(1R)-1-{[(2S,3R)-3-hydroxy-2-{[(6-phenyl-2-pyridinyl)carbonyl]amino}butanoyl]amino}-3-methylbutyl]boronic acid, is a novel proteasome inhibitor, now under early clinical evaluation as an anticancer agent. To investigate its clinical pharmacokinetics, a high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method was developed and validated to measure the drug in human plasma, based on simple protein precipitation with acetonitrile after the addition of irbesartan as internal standard. The method requires a small volume of sample (100 µl) and is rapid and selective, allowing good resolution of peaks in 5 min. It is sensitive, precise and accurate, with overall precision, expressed as coefficient of variation (CV%), always < 10.0%, accuracy in the range 93.8-107.7% and high recovery, close to 100%. The limit of detection is 0.01 ng/ml and the lower limit of quantitation (LLOQ) is 0.20 ng/ml. The assay was validated in the range from the LLOQ up to 50.00 ng/ml. This is the first method developed and validated for analyzing a proteasome inhibitor with a boronic-acid-based structure in human plasma. The method was successfully applied to study the pharmacokinetics of CEP-18770 in cancer patients with solid tumors or multiple myeloma who had received the drug as a short intravenous bolus during the initial Phase I trial.

The gut takes nearly all: threonine kinetics in infants.

BACKGROUND: Threonine is an essential amino acid that is abundantly present in intestinally produced glycoproteins. Animal studies show that intestinal first-pass threonine metabolism is high, particularly during a restricted enteral protein intake. OBJECTIVE: The objective of the study was to quantify intestinal first-pass threonine metabolism in preterm infants during full enteral feeding and during restricted enteral intake. DESIGN: Eight preterm infants (x +/- SD birth weight: 1.1 +/- 0.1 kg; gestational age: 29 +/- 2 wk) were studied during 2 periods. During period A, 40% of total intake was administered enterally and 60% was administered parenterally. Total threonine intake was 58 +/- 6 micromol kg(-1) h(-1). During period B, the infants received full enteral feeding, and the total threonine intake was 63 +/- 6 micromol kg(-1) h(-1). Dual stable-isotope tracer techniques were used to assess splanchnic and whole-body threonine kinetics. RESULTS: The fractional first-pass threonine uptake by the intestine was remarkably high in both periods: 82 +/- 6% during partial enteral feeding and 70 +/- 6% during full enteral feeding. Net threonine retention was not affected by the route of feeding. CONCLUSION: In preterm infants, the splanchnic tissues extract a very large amount of the dietary threonine intake, which indicates a high obligatory visceral need for threonine, presumably for the purposes of synthesis.

Maintenance threonine requirement and efficiency of its use for accretion of whole-body threonine and protein in Atlantic salmon (Salmo salar L.) fry.

Eighteen groups of seventy Atlantic salmon (Salmo salar L.) fry (initial mean body weight 0.8 (sd 0.01) g) were fed on semi-purified diets containing graded levels of l-threonine (Thr) in 15 litres aquaria at a temperature of 14.5+/-1 degrees C. Doses of Thr represented 1, 31, 41, 51, 62, 72, 83 and 93 % of its ideal level for optimum protein deposition. Indispensable amino acids other than Thr were included in the same proportion (on a g/16 g N basis) as in the Atlantic salmon fry whole-body carcass. Following 36 d of feeding and a 36 h fast, fry were killed for whole-body protein and amino acid analysis. Weight gain (r2 0.98), protein accretion (r2 0.97), and Thr accretion (r2 0.97) were linear (P<0.01) functions of Thr intake. Slope of the Thr accretion regression line showed that the efficiency of Thr utilisation above maintenance was 76 %. At zero Thr intake, fry lost 5.4 mg Thr/kg body weight0.75 per d. The Thr maintenance requirement was 7.2 mg/kg body weight0.75 per d and the Thr requirement for growth was 66 mg for 1 g protein deposition. Increasing doses of Thr resulted in increased (P<0.05) concentrations of histidine and lysine, and decreased concentrations of isoleucine in whole-body protein. The maintenance need for Thr represented 13.4 % of the total need for Thr. The data suggest that efficiency of Thr utilisation above maintenance is constant at all levels of Thr intake between 1 and 93 % of the level required for optimum protein deposition.

Difference in rates of net portal absorption between crystalline and protein-bound lysine and threonine in growing pigs fed once daily.

Net portal absorption of AA during the 6-h postprandial period was measured in eight gilts (48.5 +/- 1.6 kg BW) in a crossover design. The pigs had chronic catheters placed in the portal vein, carotid artery, and ileal vein, and were trained to consume 1.2 kg of a standard grower diet once daily. Blood samples were taken every 30 min for 4 h and then hourly until 6 h after feeding. The first set of blood samples was taken after pigs were fed a meal of the test 16% CP corn-soybean meal diet (16% CP) or the test 12% CP corn-soybean meal diet supplemented with crystalline lysine, threonine, and tryptophan (12% CP + AA) to equal the three AA levels in the 16% CP diet. Pigs were then fed the standard diet for 2 d. Following that, blood samples were again taken after the pigs were fed a meal of the test diet that was not given to them at the first sampling period. Net portal AA absorption was calculated by multiplying porto-arterial plasma AA concentration difference by portal vein plasma flow rate (PVPF), estimated by an indicator-dilution technique employing p-aminohippuric acid as the indicator infused into the ileal vein. Plasma concentrations of lysine and threonine of pigs were affected by the diet x time interaction (P < 0.01). Portal and arterial plasma lysine and threonine concentrations in pigs attained the maximal level by 1 h postprandial when the 12% CP + AA diet was fed, but reached the peak level at 2.5 h postprandial when the 16% CP diet was given. The PVPF of pigs over the 6 h postprandial was less (P < 0.01) when the 12% CP + AA diet was given than when the 16% CP diet was fed. Net portal absorptions of lysine and threonine also were affected (P < 0.05) by time x diet interaction. The peak portal absorption of both lysine and threonine in pigs appeared at 0.5 h postprandial when the 12% CP + AA diet was given, but at 2.5 h postprandial with the feeding of the 16% CP diet. The early appearance of peak portal absorption of lysine and threonine from feeding the 12% CP + AA compared with the 16% CP diet indicates that crystalline lysine and threonine are absorbed more rapidly than protein-bound lysine and threonine in pigs fed once daily.

Umbilical threonine uptake during maternal threonine infusion in sheep.

OBJECTIVE: The infusion into the maternal circulation of amino acid solutions failed to increase umbilical threonine (THR) uptake above normal even when THR was present in the infusate at a relatively high concentration. The purpose of the present study was to determine whether umbilical THR uptake can be increased by infusing a THR solution that does not contain any other amino acids. STUDY DESIGN: Five pregnant sheep (130+/-1.0 days after conception) were infused for 2h with a threonine solution (4.4+/-0.2 micromol.kg(-1).min(-1)). Plasma amino acids, glucose and lactate, hematocrit, blood O(2) content in maternal arterial, uterine venous, umbilical arterial and venous blood were measured. Uterine and umbilical blood flows were measured before and during the infusion and were used to calculate uterine and umbilical uptakes. Maternal and foetal plasma insulin and glucagon concentrations were also measured. RESULTS: The THR infusion increased maternal plasma THR (904 vs 236 microM, P< 0.001), foetal plasma THR (539 vs 334 microM, P< 0.01), and both uterine (20.4 vs 4.7 micromol.min(-1).kg(-1)(fetalweight), P< 0.05) and umbilical (8.6 vs 3.8 micromol.min(-1).kg(-1)(fetalweight), P< 0.001) THR uptakes. The uterine-umbilical THR uptake difference increased significantly (11.8 vs 0.9 micromol.min(-1).kg(-1)(fetalweight), P< 0.05). There were significant (P< 0.001) decreases in the foetal arterial plasma concentrations of tyrosine and the branched chain amino acids, as well as in isoleucine umbilical uptake (P< 0.05). There was a significant increase in maternal plasma glucagon (P< 0.01). CONCLUSION: A maternal THR infusion that causes a 3.8-fold increase in maternal plasma THR concentration above normal, with no significant increase in the concentration of other amino acids, leads to a 2.3-fold increase in umbilical THR uptake. This contrasts with the absence of a significant increase in umbilical THR uptake when THR was infused as part of an amino acid mixture in previous studies. The evidence supports the hypothesis that, in vivo, THR flux from placenta to foetus is mediated by a saturable, rate limiting transport system which is subject to inhibition by other neutral amino acids.

The simultaneous biosynthesis and uptake of amino acids by Lactococcus lactis studied by (13)C-labeling experiments.

Uniformly (13)C labeled glucose was fed to a lactic acid bacterium growing on a defined medium supplemented with all proteinogenic amino acids except glutamate. Aspartate stemming from the protein pool and from the extracellular medium was enriched with (13)C disclosing a substantial de novo biosynthesis of this amino acid simultaneous to its uptake from the growth medium and a rapid exchange flux of aspartate over the cellular membrane. Phenylalanine, alanine, and threonine were also synthesized de novo in spite of their presence in the growth medium.

Estimation of apparent L-amino acid diffusion in porcine jejunal enterocyte brush border membrane vesicles.

There is an overlap of carrier-mediated L-amino acid transport and apparent simple diffusion when measured in intestinal brush border membrane vesicles. Using L-threonine and L-glutamine as representative amino acids, this study was undertaken to estimate apparent simple diffusion of L-amino acids and to establish the effective dosage of HgCl2 for completely blocking carrier-mediated L-amino acid transport in porcine jejunal enterocyte brush border membrane vesicles. Jejunal mucosa was scraped from three pigs weighing 26 kg. Enterocyte brush border membrane vesicles, with an average enrichment of 24-fold in sucrase specific activity, were prepared by Mg2+-precipitation and differential centrifugation. In vitro uptake was measured by the fast filtration manual procedure. HgCl2 blocked the carrier-mediated initial transport of L-threonine and L-glutamine under Na+-gradient condition in a dose-dependent manner. At the minimal concentration of 0.165 micromol HgCl2 mg(-1) protein, carrier-mediated L-threonine and L-glutamine transport was completely inhibited. The apparent L-threonine and L-glutamine diffusion was estimated to be 8.6+/-0.7 and 12.4+/-1.0% of the total uptake at the substrate concentrations of 5 microM (L-threonine) and 50 microM (L-glutamine). Therefore, the treatment of porcine brush border membrane vesicles with a minimum of 0.165 micromol HgCl2 mg(-1) protein completely blocks carrier-mediated L-amino acid transport and enables the direct estimation of apparent L-amino acid diffusion in enterocyte brush border membrane vesicles.

An in vivo study of ovine placental transport of essential amino acids.

Under normal physiological conditions, essential amino acids (EA) are transported from mother to fetus at different rates. The mechanisms underlying these differences include the expression of several amino acid transport systems in the placenta and the regulation of EA concentrations in maternal and fetal plasma. To study the relation of EA transplacental flux to maternal plasma concentration, isotopes of EA were injected into the circulation of pregnant ewes. Measurements of concentration and molar enrichment in maternal and fetal plasma and of umbilical plasma flow were used to calculate the ratio of transplacental pulse flux to maternal concentration (clearance) for each EA. Five EA (Met, Phe, Leu, Ile, and Val) had relatively high and similar clearances and were followed, in order of decreasing clearance, by Trp, Thr, His, and Lys. The five high-clearance EA showed strong correlation (r(2) = 0.98) between the pulse flux and maternal concentration. The study suggests that five of the nine EA have similar affinity for a rate-limiting placental transport system that mediates rapid flux from mother to fetus, and that differences in transport rates within this group of EA are determined primarily by differences in maternal plasma concentration.

The effects of tannins on nutrient utilisation in the White Pekin duck.

1. Two experiments were conducted to determine the effects of tannins on nutrient utilisation in the White Pekin duck. 2. Experiment 1 was a rapid nutrient balance assay to determine the nitrogen (N) retention and metabolisable energy (ME) of maize, low-tannin sorghum (P-954063) (Sorghum bicolor (L). Moench) and high-tannin sorghum (IS-4225) cultivars for ducks. The assay lasted 120 h, with an initial 24 h food-deprivation period, a 48 h excreta collection period for endogenous losses and a 48 h excreta collection period for ingredient losses. The true metabolisable energy (TMEn) content was lower (P<0.05) in the high-tannin sorghum cultivar (13.85 MJ/kg) than the maize (14.94 MJ/kg) and the low-tannin sorghum cultivar (14.39 MJ/kg). True N retention was lower (P<0.05) for the high-tannin sorghum (0.24 g) than for maize (1.33 g) and low-tannin sorghum (1.1 g). 3. In experiment 2, the brush-border membrane vesicles technique was used to determine whether tannic acid caused inhibition of L-threonine transport across duck intestinal brush-border membrane. The brush-border membrane vesicles were mixed with tannic acid solutions (pH 7.4) to give gradient tannic acid concentrations of 0, 0.05, 0.10, 0.25, 0.50, 1.00 and 2.50%. As a fraction of the control (no tannic acid), the maximal inhibition of L-threonine transport (Imax) under the sodium-gradient condition was 77.10% (P<0.05). Under the sodium-free condition, the maximal inhibition of L-threonine transport (Imax) was 45.15% (P<0.05). 4. These results demonstrated that nutrient utilisation in the White Pekin duck was lower from the high-tannin sorghum cultivar than from the low-tannin sorghum cultivar. The results also suggested that the antinutritive effects of tannins in foodstuffs are due partly to their inhibitory action on intestinal brush-border bound amino acid transporter proteins.

Threonine kinetics in preterm infants fed their mothers' milk or formula with various ratios of whey to casein.

BACKGROUND: Plasma threonine concentrations are elevated in infants fed formula containing a whey-to-casein protein ratio of 60:40 compared with concentrations in infants fed formula containing a ratio of 20:80 or human milk (60:40). OBJECTIVE: We studied whether degradation of excess threonine was lower in formula-fed infants than in infants fed their mothers' milk. DESIGN: Threonine kinetics were examined in 17 preterm infants (gestational age: 31+/-2 wk: birth weight: 1720+/-330 g) by using an 18-h oral infusion of [1-13C]threonine at a postnatal age of 21+/-11 d and weight of 1971+/-270 g. Five infants received breast milk. Formula-fed infants (n = 12) were randomly assigned to receive 1 of 3 formulas (5.3 g protein/MJ) that differed only in the whey-to-casein ratio (20:80, 40:60, and 60:40). RESULTS: Threonine intake increased significantly in formula-fed infants with increasing whey content of the formula (48.5, 56.4, and 63.2 micromol.kg(-1).h(-1), respectively; pooled SD: 2.2; P = 0.0001), as did plasma threonine concentrations (228, 344, and 419 micromol/L, respectively; pooled SD: 75; P = 0.03). Despite a generous threonine intake by infants fed breast milk (58.0+/-16.0 micromol.kg(-1).h(-1), plasma threonine concentrations remained low (208+/-41 micromol/L). Fecal threonine excretion and net threonine tissue gain, estimated by nitrogen balance, did not differ significantly among groups. Threonine oxidation did not differ significantly among formula-fed infants but was significantly lower in formula-fed infants fed than in infants fed breast milk (17.1% compared with 24.3% of threonine intake, respectively). CONCLUSION: Formula-fed infants have a lower capacity to oxidize threonine than do infants fed breast milk.

Dietary cystine and liver triacylglycerols in rats: effects of dietary lysine and threonine.

Addition of excess cystine to a wheat gluten diet did not alter rat liver triacylglycerols or serum cholesterol. However, if the cystine-enriched diet was supplemented with lysine and threonine, rats accumulate triacylglycerols and show increased serum cholesterol. Increases in hepatic triacylglycerols can be prevented by the further addition of methionine. This diet further increases serum cholesterol. We conclude that accumulation of triacylglycerols in the liver might be due to an increased methionine requirement, induced by the addition of excess cystine, and therefore to choline deficiency.

Hyperproduction of L-threonine by an Escherichia coli mutant with impaired L-threonine uptake.

An efficient production strain for L-threonine fermentation was derived from Escherichia coli by multiple rounds of mutation programs that aimed at deregulation of the L-threonine biosynthetic pathway and blocking of L-threonine degradation pathways. When the optimum amount of DL-methionine was added, this strain KY10935, an L-methionine auxotroph, gave 100 g/liter L-threonine after 77 h cultivation. In this strain, key enzymes in the L-threonine biosynthetic pathway were highly derepressed, but some were inhibited by lower concentrations of L-threonine than the accumulated level. Such incomplete deregulation of the pathway was accounted for by the intracellular concentration of L-threonine being lower than the extracellular level. In an assessment of L-threonine transport in terms of phenotypic growth responses to the amino acid, L-threonine-auxotrophic mutants with a lesion in the L-threonine operon were derived from strain KY10935 by selection for auxotrophy for dipeptide L-alanyl-L-threonine or glycyl-L-threonine, the transport systems of which were different from those of L-threonine. All three independent mutants isolated needed an extraordinarily high concentration (10 mg/ml) of L-threonine, but grew in the presence of a low concentration (10 micrograms/ml) of either dipeptide, indicating that strain KY10935 had impaired L-threonine uptake. These results suggested that the strain had an unusual mechanism of L-threonine hyperproduction: the inability to take up L-threonine that had accumulated extracellularly decreased the steady-state level of intracellular L-threonine, freeing the remaining regulatory steps of feedback inhibition.

Effects of histamine on mucin biosynthesis in rat gastric mucosa.

Mucin biosynthesis is stimulated by gastrin during the process of glycosylation in the corpus mucosa of the rat stomach. The purpose of this study was to clarify, using an organ culture technique, whether biosynthetic responses to histamine in the rat gastric mucin are the same as that to gastrin. Radiolabeled mucin was obtained from the corpus and antral mucosa of the rat stomach after in vitro incubation for 5 h with [3H]glucosamine (GlcN), [14C]threonine (Thr), and [35S]sulfate. Addition of histamine (10(-7)-10(-5) M) to the culture medium increased [3H]GlcN-labeled mucin in the corpus tissue in a concentration-dependent manner. In the antrum, there was no significant change in the biosynthetic activity of mucin in response to histamine. Histamine at 10(-5) M also increased the incorporation of both [35S]sulfate and [14C]Thr into the corpus mucin. These results indicate that histamine stimulates the biosynthesis of the mucin peptide, as well as the glycosylation step in the corpus, and suggest that the effect of histamine on mucin synthesis is distinct from that of gastrin.

Inhibition of L-threonine intestinal absorption in rabbits by cadmium.

Cadmium compounds are widely spread in the environment. Animal exposure to cadmium compounds occurs mainly through foods or drinks contaminated by this metal. Cadmium has been shown to produce several negative effects on the gastrointestinal tract such as inhibition on sugars and amino acids absorption. The aim of the present work was to study the inhibitory characteristics of cadmium on L-threonine intestinal absorption in rabbits in order to understand about this malabsorption of nutrients. Our results show that L-threonine tissue accumulation as well as mucosal to serosal transepithelial fluxes are decreased in a dose-dependent manner in rabbit jejunum. Amino acid diffusion across the intestinal epithelium was not affected by cadmium. A noncompetitive mechanism and a partial reversion by dithioerythritol (thiol groups protector) is described for this inhibition.

Bioavailability of threonine in soybean meal for young pigs.

The bioavailability of threonine in solvent-extracted soybean meal for 10- to 20-kg pigs was determined using the slope-ratio method. In Exp. 1, the assay range was determined by feeding six diets to 144 pigs. The basal diet (.40% threonine) contained corn, soybean meal, and corn gluten meal. Five additional diets were formulated by supplementing the basal diet with .05 to .25% crystalline L-threonine in .05% increments. Weight gain, gain/feed, and plasma concentrations of threonine and urea responded quadratically (P < .05) to increasing dietary threonine. Breakpoints ranged from .51 to .54% dietary threonine. Experiment 2 consisted of seven trials in which a total of 239 pigs were used in a randomized complete block design. Pigs were penned individually and had ad libitum access to feed and water during the 21-d experiment. The same basal diet that was used in Exp. 1 was supplemented with .018, .053, or .070% threonine from either L-threonine or soybean meal. The weight gains of the pigs were partitioned to yield the response due to the supplemental threonine ingested. Multiple regression was performed on partitioned weight gain vs supplemental threonine intake, and the assay was tested for validity. The regression lines for L-threonine and soybean meal were linear (P < .05) and the intercepts were not significantly different (P > .10). The slope ratio for soybean meal:L-threonine was .80. Although the difference between the soybean meal and L-threonine slopes was not significant (P > .23), the best estimate of the bioavailability of threonine in soybean meal relative to that of L-threonine was 80%.

Effect of erythromycin on L-threonine transport in rabbit jejunum in vitro.

Several antibiotics characterized by different molecular structures are known to affect some intestinal activities. Some of them have been described as inhibitors of the intestinal sugar and amino acid transport with different mechanisms. Erythromycin (EM) is a macrolide antibiotic acting as a motilin agonist and thus stimulating the gastrointestinal motor activity. Since several substances which increase the motor activity of the gastrointestinal tract may produce effects on the intestinal absorption of nutrients, the present study has been carried out to determine whether erythromycin affects the L-threonine intestinal absorption. The results obtained indicate that erythromycin diminishes the L-threonine intestinal transport, probably at the mucosal border level. Two groups of experiments carried out, with Na(+)-deprived medium and ouabain-enriched medium, might indicate that erythromycin action could be due to either a direct or an indirect action on the Na(+)-dependent L-threonine transport located in the brush border.

Bioavailability of threonine in soybean meal for rats and chicks.

The bioavailability of threonine in soybean meal and the effects of the excess amino acids in soybean meal on the estimate were measured using rats and chicks in slope-ratio assays. In Exp. 1, a corn-based diet containing .23% threonine was supplemented with 0 to 45% L-threonine in .05% increments. The growth rate of weanling rats fed these diets increased quadratically (P less than .001) with L-threonine addition, the increase being essentially linear up to the .10% addition. In Exp. 2, the basal diet was supplemented with 0, .025, .050, .075, or 100% threonine from L-threonine, simulated soybean meal (a mixture of crystalline amino acids with a pattern designed to simulate soybean meal), or soybean meal. Regressions of partitioned weight gain and body N gain of rats vs supplemental threonine intake were calculated for each source using multiple regression. Slope ratios (soybean meal:L-threonine) were .91 for weight gain and .92 for body N gain. The additional amino acids in simulated soybean meal did not affect the estimate. For Exp. 3, a corn-soybean meal-based diet containing .48% threonine was supplemented with 0 to 60% L-threonine in .10% increments. The growth rate of broiler chicks fed the diets increased quadratically (P less than .001) with L-threonine addition. The increase was essentially linear up to the .10% addition. In Exp. 4, the basal diet was supplemented as in Exp. 2. Regressions of partitioned weight gain of chicks vs supplemental threonine intake were calculated for each source. The slope ratio for soybean meal:L-threonine was 1.03; however, the model exhibited fundamental invalidity and therefore the estimate should be interpreted with caution. The additional amino acids in the simulated soybean meal did not affect the value.