Supplementation of Sulfur-Containing Amino Acids or Essential Amino Acids Does Not Reverse the Hepatic Lipid-Lowering Effect of a Protein-Rich Insect Meal in Obese Zucker Rats.

The present study tested the hypothesis that the liver lipid-lowering effect of insect meal (IM) is caused by its low methionine concentration. A total of fifty, male obese Zucker rats were randomly assigned to five groups of 10 rats each (casein (C), IM, IM + Met, IM + Cys, and IM + EAA). While group C received a diet with casein, the IM-fed groups received a diet with IM as the protein source. In groups IM + Met, IM + Cys and IM + EAA, the diets were additionally supplemented with methionine, cysteine and essential amino acids (EAA), respectively. Hepatic concentrations of triacylglycerols and cholesterol, and hepatic mRNA levels and activities of lipogenic and cholesterogenic enzymes were markedly lower in the IM-fed groups than in group C (p < 0.05). All of these parameters either did not differ across the IM-fed groups or were only slightly higher in groups IM + Met, IM + Cys and IM+EAA than in the group IM. In conclusion, the results indicate that a difference in the amino acid composition between IM and casein, a low concentration of methionine in IM and a reduced cysteine synthesis secondary to a decreased methionine availability resulting from feeding IM are not causative for the lipid-lowering effect of IM.

Substitution of Dietary Sulfur Amino Acids by dl-2-Hydroxy-4-Methylthiobutyric Acid Reduces Fractional Glutathione Synthesis in Weaned Piglets.

BACKGROUND: Cys is limiting for reduced glutathione (GSH) synthesis and can be synthesized from Met. We hypothesized that the dietary Met hydroxyl analogue dl-2-hydroxy-4-methylthiobutyric acid (dl-HMTBA) affects Cys and GSH metabolism and oxidative stress defense differently than Met. OBJECTIVE: The objective was to elucidate whether dl-HMTBA supplementation of a Met-deficient diet affects Cys flux, GSH fractional synthetic rate (FSR), and the basal oxidative stress level relative to Met supplementation in pigs. METHODS: Twenty-nine male German Landrace piglets aged 28 d were allocated to 3 dietary groups: a basal diet limiting in Met (69% of Met plus Cys requirement) supplemented with either 0.15% l-Met (LMET; n = 9), 0.15% dl-Met (DLMET; n = 11), or 0.17% dl-HMTBA (DLHMTBA; n = 9) on an equimolar basis. At age 54 d the pigs received a continuous infusion of [1-13C]-Cys to calculate Cys flux and Cys oxidation. After 3 d, GSH FSR was determined by [2,2-2H2]-glycine infusion, and RBC GSH and oxidized GSH concentrations were measured. At age 62 d the animals were killed to determine hepatic mRNA abundances of enzymes involved in GSH metabolism, GSH concentrations, and plasma oxidative stress defense markers. RESULTS: The Cys oxidation was 21-39% and Cys flux 5-15% higher in the fed relative to the feed-deprived state (P < 0.001). On average, GSH FSR was 49% lower (P < 0.01), and RBC GSH and total GSH concentrations were 12% and 9% lower, respectively, in DLHMTBA and DLMET relative to LMET pigs (P < 0.05). In the feed-deprived state, Gly flux, the GSH:oxidized glutathione (GSSG) ratio, RBC GSSG concentrations, plasma oxidative stress markers, and the hepatic GSH content did not differ between groups. CONCLUSIONS: Although GSH FSR was higher in LMET compared with DLMET or DLHMTBA feed-deprived pigs, these differences were not reflected by lower oxidative stress markers and antioxidant defense enzymes in LMET pigs.

Changes in cecum microbial community in response to total sulfur amino acid (TSAA: DL-methionine) in antibiotic-free and supplemented poultry birds.

The effect of essential total sulfur amino acids (TSAA) like methionine and cysteine on the cecal microbiome of broilers was investigated at 2 different time points (days 21 and 42) of broiler rearing. A total of 360-day-old Cobb male broiler chicks were randomly distributed to 6 dietary treatments in a 2 × 3 factorial arrangement, with 2 levels of antibiotic growth promoters (AGP: 0 and 0.05%) and 3 levels of TSAA (DL-methionine) either for starter (0.7, 0.8, and 0.9%) or finisher chicks (0.52, 0.62, and 0.72%), labeled as diets 1 to 6. Cecal digesta from each replicate (n = 10) were sampled on days 21 and 42. DNA was extracted for the amplification of the V4 region of bacterial 16S rRNA genes and subjected to Illumina sequencing. Bioinformatic analyses were performed using QIIME, Mothur, and ad hoc tools and functional profiles of the inferred metagenome were analyzed using PICRUST. Statistical difference was determined by 2-way ANOVA and PERMANOVA. Clustering of cecal communities using PCoA showed clear separation of microbial communities based on age (P < 0.05) of birds and between low and medium/ high levels of TSAA (DL-methionine). At day 21, bacterial richness and diversity were higher than at day 42 where Clostridium cluster XI and Lactobacillus were found most abundant. No variability in taxonomic richness at the genus level was observed with AGP and DL-methionine supplementation. Interbird variation for richness was greater at day 42 compared to day 21. The mean fold difference of richness was greater (1.5 mean fold) with diets 1 and 6, suggesting interactive effects of AGP and TSAA (DL-methionine) in the diet. KEGG function profiles calculated by PICRUST suggest that the cecal microbiome increased glycolysis and energy generation correlated with increased dietary TSAA (DL-methionine) supplementation levels during the late broiler growth period (day 42). This study increases our knowledge of microbial dynamics and functions that are relevant to host nutrition and performance that may help us tailoring alternative strategies for raising poultry birds under antibiotic-free conditions.

Estimation of the standardized ileal digestible lysine requirement and optimal sulphur amino acids to lysine ratio for 30-50 kg pigs.

Two experiments were conducted to determine the standardized ileal digestible (SID) lysine (Lys) requirement and the ideal SID sulphur amino acids (SAA) to Lys ratio for 30-50 kg crossbred pigs. In experiment 1, a total of 72 crossbred pigs with an average initial body weight (BW) of 28.9 kg were allotted to one of six dietary treatments in a randomized complete block design. Each diet was assigned to six pens containing two pigs each. Six diets were obtained by supplementing graded levels of L-Lysine∙HCl to create six dietary levels of SID Lys (0.70%, 0.80%, 0.90%, 1.00%, 1.10% and 1.20%). Responses of weight gain (ADG) and gain:feed (G:F) to increasing the SID Lys content of the diet fitted well with the curvilinear-plateau model; whereas, for plasma urea nitrogen (PUN) two-slope linear broken-line model was well fitted. The optimal SID Lys requirement for the pigs of this period was 1.10%. Experiment 2 was a dose-response study using SID Met+Cys to Lys ratios of 50%, 55%, 60%, 65%, 70% and 64%. A total of 72 crossbred pigs with initial BW of 32.9 kg were randomly allotted to receive one of the six diets. Diets 1-5 were formulated to contain 1.0% SID Lys to be second limiting in Lys and diet 6 contained 1.11% SID Lys to be adequate in Lys. The average optimal SID SAA:Lys ratio for maximal ADG and G:F and minimal PUN was 65.2% using curvilinear-plateau and linear broken-line models.

Dietary sulfur amino acids affect jejunal cell proliferation and functions by affecting antioxidant capacity, Wnt/ß-catenin, and the mechanistic target of rapamycin signaling pathways in weaning piglets.

Intestinal epithelial cells undergo rapid renewal along the crypt-villus axis (CVA), which ensures intestinal functions. Weaning stress differentially effects intestinal epithelial cell metabolism and physiological states along the CVA. Sulfur amino acids (SAA) play a key role in intestinal epithelial cell functioning. This study evaluated the effects of SAA dietary supplementation on weaning pig jejunal epithelial cells along the CVA. Sixteen Duroc × Landrace × Yorkshire piglets (6.16 ± 0.22 kg BW) were weaned at 21 d of age and were blocked by BW and gender and the randomly assigned to 1 of 2 groups fed diets consisting of low (0.53%) or high (0.85%) levels of SAA for a 7-d period. All piglets were euthanized for tissue sampling on day 7 postweaning. Jejunal epithelial cells were isolated along the CVA to yield 3 "cell fractions" (upper villus, middle villus, and crypt cells). The number of proliferating cells per crypt of piglets fed the high SAA diet was lower (P < 0.05) than that for low SAA diet. High SAA diet piglets tended to have decreased (P = 0.059) sucrase activities compared low SAA diet piglets. A high SAA diet increased (P < 0.05) total antioxidant capacity, catalase, and superoxide dismutase activities compared with a low SAA diet. mRNA expression levels of claudin-1, Slc5a1, and Slc7a9 in high SAA diet piglets were lower (P < 0.05) than for low SAA diet piglets. There were no interactions between dietary SAA and cell sections along the CVA for enzyme activities and mRNA expression in any of the weaned piglets. Protein amounts and phosphorylation levels related to Wnt/ß-catenin and mechanistic targeting of rapamycin (mTOR) signaling pathways were affected by SAA in weaning piglets. These findings indicate that dietary SAA affects jejunal cell proliferation and functions in weaning piglets. There appears to be no interactions between dietary SAA and cell sections along the CVA. The effects of SAA may be partly through affecting antioxidant capacity, and Wnt/ß-catenin and mTOR signaling pathway.

Food intake of selenium and sulphur amino acids in tuberculosis patients and healthy adults in Malawi.

Wasting in tuberculosis (TB) patients is associated with the severity of lung disease and low serum level of selenium. Low serum levels of selenium may be due to low food intake or altered absorption/metabolism in the body. We therefore wanted to measure and compare the intake of selenium and sulphur amino acids in smear-positive TB patients and appropriately matched adults. We found a low intake of selenium but no significant difference in intake of selenium and sulphur amino acids between the groups in this study.

Total sulfur amino acid requirement and metabolism in parenterally fed postsurgical human neonates.

BACKGROUND: Except for tyrosine, the amino acid requirements of human neonates receiving parenteral nutrition (PN) have not been experimentally derived. OBJECTIVES: The objectives were to determine the total sulfur amino acid (TSAA) requirement (methionine in the absence of cysteine) of postsurgical, PN-fed human neonates by using the indicator amino acid oxidation (IAAO) technique with L-[1-(13)C]phenylalanine as the indicator. DESIGN: Fifteen postsurgical neonates were randomly assigned to receive 1 of 18 methionine intakes ranging from 10 to 120 mg x kg(-1) x d(-1), delivered in a customized, cysteine-free amino acid solution. Breath and urine samples were collected for the measurement of (13)CO(2) and amino acid enrichment. Blood samples were collected at baseline and after the test methionine infusion for the measurement of plasma methionine, homocysteine, cystathionine, and cysteine concentrations. RESULTS: Breakpoint analysis determined the mean TSAA requirements to be 47.4 (95% CI: 38.7, 56.1) and 49.0 (95% CI: 39.9, 58.0) mg x kg(-1) x d(-1) with the use of oxidation and F(13)CO(2), respectively. CONCLUSIONS: This is the first study to report the TSAA requirement of postsurgical, PN-fed human neonates. The estimated methionine requirement expressed as a proportion of the methionine content of current commercial pediatric PN solutions was 90% (range: 48-90%) of that found in the lowest methionine-containing PN solution.

The indicator amino acid oxidation method identified limiting amino acids in two parenteral nutrition solutions in neonatal piglets.

Recent studies using the indicator amino acid oxidation (IAAO) technique in TPN-fed piglets and infants have been instrumental in defining parenteral amino acid requirements. None of the commercial products in use are ideal when assessed against these new data. Our objectives were to determine whether the oxidation of an indicator amino acid would decline with the addition of amino acids that were limiting in the diets of TPN-fed piglets, and to use this technique to identify limiting amino acids in a new amino acid profile. Piglets (n = 26) were randomized to receive TPN with amino acids provided by Vaminolact (VM) or by a new profile (NP). After 5 d of TPN administration, lysine oxidation was measured using a constant infusion of L- [1-(14)C]-lysine. Immediately following the first IAAO study, the piglets were further randomized within diet group to receive either 1) supplemental aromatic amino acids (AAA), 2) sulfur amino acids (SAA) or 3) both (AAA+SAA) (n = 4-5 per treatment group). A second IAAO study was carried out 18 h later. In the first IAAO study, lysine oxidation was high for both groups (18 vs. 21% for VM and NP, respectively, P = 0.055). The addition of AAA to VM induced a 30% decline in lysine oxidation compared with baseline (P < 0.01). Similarly, SAA added to NP lowered lysine oxidation by approximately 30% (P < 0.01). The application of the IAAO technique facilitates rapid evaluation of the amino acids that are limiting to protein synthesis in parenteral solutions.

High hepatic glutathione stores alleviate Fas-induced apoptosis in mice.

BACKGROUND/AIMS: The agonistic Jo2 anti-Fas antibody reproduces human fulminant hepatitis in mice. We tested the hypothesis that enhancing hepatic glutathione (GSH) stores may prevent Jo2-induced apoptosis. METHODS: We fed mice with a normal diet or a sulfur amino acid-enriched (SAA(+)) diet increasing hepatic GSH by 63%, and challenged these mice with Jo2. RESULTS: The SAA(+) diet markedly attenuated the Jo2-mediated decrease in hepatic GSH and the increase in the oxidized glutathione (GSSG)/GSH ratio in cytosol and mitochondria. The SAA(+) diet prevented protein kinase Czeta (PKCzeta) and p47(phox) phosphorylations, Yes activation, Fas-tyrosine phosphorylation, Bid truncation, Bax, and cytochrome c translocations, the mitochondrial membrane potential collapse, caspase activation, DNA fragmentation, hepatocyte apoptosis, and mouse lethality after Jo2 administration. The protective effect of the SAA(+) diet was abolished by a small dose of phorone decreasing hepatic GSH back to the levels observed in mice fed the normal diet. Conversely, administration of GSH monoethyl ester after Jo2 administration prevented hepatic GSH depletion and attenuated toxicity in mice fed with the normal diet. CONCLUSIONS: The SAA(+) diet preserves GSSG/GSH ratios, and prevents PKCzeta and p47(phox) phosphorylations, Yes activation, Fas-tyrosine phosphorylation, mitochondrial permeabilization, and hepatic apoptosis after Fas stimulation. GSH monoethyl ester is also protective, suggesting possible clinical applications.

The effects of sulfur amino acid intake on immune function in humans.

No direct data exist on the influence of supranormal intakes of sulfur amino acids on immune function in humans. However 3 major products of sulfur amino acids, glutathione (GSH), homocysteine (Hcy), and taurine (Tau), influence, mainly, inflammatory aspects of the immune response in vitro and in vivo. Methionine intakes above approximately 1 g/d transiently raise plasma Tau, Hcy, and GSH. Tau and GSH ameliorate inflammation. Hcy has the opposite effect. A biphasic relation, between cellular GSH and CD4+ and CD8+ numbers occurs in healthy men. How changes in sulfur amino acid intake influence this phenomenon is unknown. In animals, high Tau intakes are antiinflammatory. How immune function in humans is affected is unknown. A positive relation between plasma neopterin (a marker of a Th-1-type immune response) and Hcy indicates that Hcy may play a part in inflammatory aspects of Parkinson's disease and aging. In vitro, Hcy, at concentrations seen following consumption of approximately 6 g L-methionine/d in adults, increases the interactions among T lymphocytes, monocytes, and endothelium. Whether a similar phenomenon occurs in vivo is unknown. Polymorphisms in the methylenetetrahydrofolate reductase gene are associated with raised plasma Hcy in young but not old subjects. The relation of this observation to immune function is unknown. The relationships among Hcy, inflammatory aspects of disease, and in vitro alterations in immune cell behavior create a cautionary note about supplementation of diets with l-methionine to raise intake above approximately 1 g/d. Studies directly linking methionine intake, genetics, plasma Hcy, Tau, and GSH and immune function are needed.

Adequate range for sulfur-containing amino acids and biomarkers for their excess: lessons from enteral and parenteral nutrition.

The adequacy range of dietary requirements of specific amino acids in disease states is difficult to determine. In health, several techniques are available allowing rather precise quantification of requirements based on growth of the organism, rises in plasma concentration, or increases in the oxidation of marker amino acids during incremental administration of the amino acid under study. Requirements may not be similar in disease with regard to protein synthesis or with regard to specific functions such as scavenging of reactive oxygen species by compounds including glutathione. Requirements for this purpose can be assessed only when such a function can be measured and related to clinical outcome. There is apparent consensus concerning normal sulfur amino acid (SAA) requirements. WHO recommendations amount to 13 mg/kg per 24 h in healthy adults. This amount is roughly doubled in artificial nutrition regimens. In disease or after trauma, requirements may be altered for methionine, cysteine, and taurine. Although in specific cases of congenital enzyme deficiency, prematurity, or diminished liver function, hypermethionemia or hyperhomocysteinemia may occur, SAA supplementation can be considered safe in amounts exceeding 2-3 times the minimal recommended daily intake. Apart from some very specific indications (e.g., acetaminophen poisoning), the usefulness of SAA supplementation is not yet established. There is a growing body of data pointing out the potential importance of oxidative stress and resulting changes in redox state in numerous diseases including sepsis, chronic inflammation, cancer, AIDS/HIV, and aging. These observations warrant continued attention for the potential role of SAA supplementation. In particular, N-acetylcysteine remains promising for these conditions.

Effect of potassium salts in rats adapted to an acidogenic high-sulfur amino acid diet.

Low-grade metabolic acidosis, consecutive to excessive catabolism of sulfur amino acids and a high dietary Na:K ratio, is a common feature of Western food habits. This metabolic alteration may exert various adverse physiological effects, especially on bone, muscle and kidneys. To assess the actual effects of various K salts, a model of the Westernised diet has been developed in rats: slight protein excess (20 % casein); cations provided as non-alkalinising salts; high Na:K ratio. This diet resulted in acidic urine (pH 5.5) together with a high rate of divalent cation excretion in urine, especially Mg. Compared with controls, K supplementation as KCl accentuated Ca excretion, whereas potassium bicarbonate or malate reduced Mg and Ca excretion and alkalinised urine pH (up to 8). In parallel, citraturia was strongly increased, together with 2-ketoglutarate excretion, by potassium bicarbonate or malate in the diet. Basal sulfate excretion, in the range of 1 mmol/d, was slightly enhanced in rats fed the potassium malate diet. The present model of low-grade metabolic acidosis indicates that potassium malate may be as effective as KHCO3 to counteract urine acidification, to limit divalent cation excretion and to ensure high citrate concentration in urine.

Responses of growing broilers to diets with increased sulfur amino acids to lysine ratios at two dietary protein levels.

An experiment with 1,440 male Cobb 500 and 1,440 male Ross 308 broilers (14 to 35 d of age) was conducted to investigate the effects of diets having 4 levels of digestible methionine plus cysteine (SAA) on various performance criteria at 2 dietary protein levels (20.5 and 26.0%). Two corn-soybean meal/poultry by-product basal diets were formulated to contain 3,060 kcal/kg MEn and either 20.5 or 26.0% balanced protein, and 1.12 and 1.46% digestible (according to table values) lysine, respectively. Except for SAA, the ratios between essential amino acids were kept identical in both diets according to the ideal protein concept. The ratio between digestible SAA and digestible Lys was 50%. All remaining nutrients met or exceeded NRC (1994) recommendations. Graded levels of SAA were supplemented to obtain digestible SAA to Lys ratios of 62, 69, and 77%, with 77% representing an optimized amino acid balance. Increasing the protein level clearly improved weight gain, feed conversion, breast meat yield, and abdominal fat content. Increasing SAA levels resulted in strong nonlinear or linear dose responses at both protein levels and for both strains. Regression analysis suggested that reducing digestible SAA in a balanced protein (diets with SAA:Lys of 77%) impairs performance, and that optimum SAA:Lys ratio for growing broilers might be higher than 77%, although ANOVA revealed no significant improvement with an SAA:Lys ratio higher than 69%. Responses provide evidence that optimum dietary SAA level depends on dietary protein level and should therefore be related to the protein content.

The effect of graded levels of dietary casein, with or without methionine supplementation, on glutathione concentration in unstressed and endotoxin-treated rats.

Glutathione (GSH) concentration was measured in rats fed either graded levels of dietary casein (experiment 1; 180 g, 120 g, 80 g, or 60 g protein/kg diet) or graded levels of dietary casein, supplemented with methionine to equalize dietary sulfur amino acid content to that seen in an 180 g/kg casein diet supplemented with 0.3 g L-methionine/kg diet (experiment 2; 180 g protein +0.3 g L-methionine, 80 g protein +6.70 g L-methionine, or 60 g protein +7.45 g L-methionine/kg diet). Rats were given an inflammatory challenge by intraperitoneal injection of endotoxin (lipopolysaccharide from Escherichia coli), and were compared with ad libitum and pair-fed controls. Glutathione concentration in various organs (liver, lung, spleen, and thymus) decreased in animals fed the low-protein diets (80 g or 60 g/kg diet). Addition of the sulfur amino acid, methionine, to the low-protein diets restored glutathione concentrations in animals fed ad libitum and prevented the fall in GSH concentration, which occurred in lung, spleen, and thymus in response to the endotoxin. Despite the similarity in the amount of sulfur amino acid consumed between the groups fed the 180 g protein +0.3 g L-methionine and the 60 g protein +7.45 g L-methionine/kg diet, in experiment 2, hepatic GSH concentration significantly increased in the latter group, in animals fed ad libitum and in the endotoxin-treated animals, but not in the pair-fed controls.

Influence of supplemental lysine, isoleucine, threonine, tryptophan and total sulfur amino acids on egg weight of Hy-line W-36 hens.

Four experiments were conducted to determine whether synthetic lysine, isoleucine, threonine, and tryptophan (LITT) or TSAA supplementation to diets formulated based on lysine improved egg weight (EW) and hen performance. In Experiment 1, diets were formulated with three TSAA levels (0.65,0.72, and 0.81%). These diets were fed with and without adding lysine and isoleucine each at 0.055% and threonine and tryptophan each at 0.022%. Treatments were randomly assigned to 960 Hy-Line W-36 hens in eight replicates per treatment (20 birds/replicate). Inclusion of LITT improved (P < 0.05) EW within 2 wk with no TSAA x LITT interaction. Inclusion of LITT also increased average feed consumption (FC) by 1.4 g/d (P < 0.01). However, no effect (P > 0.05) of LITT was observed on egg production (EP), egg specific gravity (SG), or BW. The EW, EP, and FC increased linearly (P < 0.05) as dietary TSAA increased. The TSAA had no effect on SG or BW. In Experiment 2, supplemental LITT were removed to determine how rapidly hens lose the effect of LITT on performance. Within 2 wk, the effect of LITT on EW was lost. Average EW, EP, and FC of hens fed higher TSAA levels remained higher (P < 0.05) than hens fed 0.65% TSAA. In Experiment 3, all hens were fed a diet containing 0.65% TSAA to determine the time it takes to lose the effect of TSAA supplementation. The TSAA supplementation effect on EW, EP, and FC observed in the previous two experiments was lost within 1 wk. In Experiment 4, hens were again fed diets similar to Experiment 2 to confirm the time required for TSAA supplementation to improve EW. Within 1 wk, hens fed 0.72 or 0.81% TSAA improved (P < 0.05) EW over hens fed 0.65% TSAA. It was concluded that hens fed diets formulated based on lysine were deficient in LITT. Hens responded to the inclusion or removal of supplemental LITT and TSAA within 1 to 2 wk.

The effects of dietary sulfur amino acid deficiency on rat brain glutathione concentration and neural damage in global hemispheric hypoxia-ischemia.

Primary brain injury in stroke is followed by an excitotoxic cascade, oxidative stress and further neural damage. Glutathione is critical and depleted in oxidative stress. Since cysteine is limiting in glutathione synthesis, this study investigated the effect of dietary sulfur amino acid (SAA) deficiency on neural damage in a rat model of global hemispheric hypoxia-ischemia (GHHI). Animals were fed with SAA deficient ("deficient") or control diet for 3 days, subjected to right common carotid artery ligation and hypoxia, and diet continued for 3 more days. Histologically evaluated neural damage at 7 days post hypoxia-ischemia was greater in "deficient" rats, shown by mean (+/- SEM) global and hippocampal grid scores of 2.5 +/- 0.7 and 34.9 +/- 9.3%, respectively, vs. controls' scores of 0.1 +/- 0.1 and 0.1 +/- 0.1%, respectively. Mean brain (+/- SEM) reduced glutathione was not different between groups at 6h post hypoxia-ischemia, but was decreased in "deficient" animals 3 days later in neocortex (1.46 micromoles/g wet weight +/- 0.05 vs. 1.67 +/- 0.04 in controls) and thalamus (1.60 micromoles/g wet weight +/- 0.05 vs. 1.78 +/- 0.03 in controls). Administration of a cysteine precursor to "deficient" animals did not ameliorate neural damage. These findings suggest that well-nourished but not "deficient" animals tolerate a mild brain insult. The decline in brain glutathione in the "deficient" animals may be one of several contributing mechanisms.

Effect of nonessential amino acids on nitrogen retention in growing pigs fed on a protein-free diet supplemented with sulphur amino acids, threonine and tryptophan.

Two N balance experiments were conducted on growing pigs to study the effect of essential and nonessential amino acids added to a protein-free diet on N retention. In Expt. 1, the addition of sulphur amino acids, threonine and tryptophan to a protein-free diet at levels two times the maintenance requirements reduced (p > 0.1) daily N loss from -131 to -108 mg/kg(0.75). A further addition of nonessential amino acids equivalent to 250 mg N/kg(0.75)/d resulted in a marked increase (p < 0.01) in daily N retention to 28 mg/kg(0.75). In Expt. 2, nonessential amino acids were added to a protein-free diet supplemented with sulphur amino acids, threonine and tryptophan at levels corresponding to 100, 200 and 300 mg N/kg(0.75)/d. N retention increased linearly as dietary nonessential N increased. The slope of the best-fit regression line indicated that the marginal efficiency of nonessential N utilization for protein accretion was 0.26. The results suggest that nonessential amino acids may be a limiting factor for the re-utilization of amino acids released by body protein breakdown or that they may serve as precursors for de novo synthesis of amino acids by gut microorganisms, thus contributing to the amino acid requirements of the pig.

Use of bioimpedance spectroscopy to estimate body water distribution in rats fed high dietary sulfur amino acids.

The effect of dietary sulfur amino acids on bioelectric properties was studied in rats by using bioimpedance spectroscopy. Weanling rats were assigned to one of 12 groups in a factorially arranged experiment with dietary variables of supplemental sulfur amino acid (none, 10 g DL-methionine/kg or 10 g DL-homocystine/kg), pyridoxine hydrochloride (0 or 7.5 mg/kg) and nickel (0 or 1 mg/kg). After 9 wk of feeding, 20-h urine specimens were collected from food-deprived rats for measurements of creatinine, and then bioimpedance was measured with multifrequency (Hydra ECF/ICF 4200) and single-frequency (RJL Systems model 101) analyzers. Urinary creatinine excretion was measured by intracellular water (ICW), total body solid and urinary volume (R2 = 0.675). Extracellular water (ECW) did not add significantly to the model. Rats fed methionine had significantly lower total body water, ICW and ECW than rats fed no supplemental sulfur amino acid. Rats fed homocystine had significantly lower ECW and a significantly higher ratio of ICW to ECW. Rats fed methionine or homocystine had significantly lower capacitance corrected for body length and ICW than those fed no supplemental sulfur amino acids. These results suggest that dietary homocystine changes the distribution of body water and that sulfur amino acids can affect membrane porosity and/or membrane thickness.

Hepatic betaine-homocysteine methyltransferase activity in the chicken is influenced by dietary intake of sulfur amino acids, choline and betaine.

There is much interest in the metabolism of homocysteine, because elevated plasma homocysteine [hyperhomocyst(e)inemia] is an independent risk factor for the development of cardiovascular disease. Four chick assays were conducted to determine the effects of varying dietary sulfur amino acids, choline and betaine on the activity of hepatic betaine-homocysteine methyltransferase (BHMT), an enzyme likely to be important in modulating plasma homocysteine. In Experiment 1, chicks were fed a purified crystalline amino acid diet containing adequate sulfur amino acids and choline. Excess dietary methionine, or the combination of excess cystine with choline or betaine, caused a small increase (P < 0.05) in BHMT activity. In Experiment 2, use of a methionine-deficient purified diet resulted in a threefold increase (P < 0.05) in BHMT activity, and addition of choline or betaine further increased (P < 0.05) BHMT activity. In Experiment 3, use of a methionine-deficient corn-peanut meal diet increased BHMT (P < 0.05) relative to that of chicks supplemented with adequate methionine, and addition of surfeit choline to the methionine-deficient basal diet caused a further increase (P < 0.05). In Experiment 4, addition of both surfeit choline and surfeit betaine to the methionine-deficient corn-peanut meal diet caused an increase (P < 0.05) in BHMT activity relative to that observed in chicks fed the methionine-deficient basal diet. These assays show that large increases in BHMT activity can be produced under methionine-deficient conditions, especially in the presence of excess choline or betaine.

Sulfur amino acid requirement and cystine replacement value of broiler chicks during the period three to six weeks posthatching.

Three experiments were conducted with commercial broiler chicks to determine the SAA requirement during the growth period 3 to 6 wk posthatching. A 20% CP corn-peanut meal basal diet (3,200 kcal ME(n)/kg) was analyzed to contain 0.23% Met and 0.28% cystine. True digestibility assessment in cecectomized cockerels revealed that Met and cystine in the basal diet were 81 and 75% digestible, respectively. Therefore, the basal diet contained 0.19% digestible Met and 0.21% digestible cystine. When fully fortified with DL-Met, growth rate and feed efficiency of chicks fed the corn-peanut meal diet were equal to that of chicks fed a 20% CP Met-fortified corn-soybean meal diet. In the SAA requirement assay, Ross x Hubbard male chicks were fed graded increments of DL-Met (0.03%) and L-cystine (0.03%) to achieve digestible SAA concentrations of 0.40, 0.46, 0.52, 0.58, 0.64, and 0.70%. Weight gain and feed efficiency responded quadratically (P < 0.01) to increasing doses of SAA. The estimated requirement for maximal feed efficiency was higher than that for maximal weight gain. Both visual appraisal and curve fitting procedures suggested a requirement of close to 0.61% digestible SAA. When extrapolated to a corn-soybean meal diet where SAA true digestibility is 87.5%, the total SAA requirement calculates to be 0.70% of the diet. However, because commercial corn-soybean meal diets typically contain supplemental Met, which is only 81% efficient (wt:wt) in furnishing cystine, the estimated total SAA requirement for chicks fed 20% CP Met-fortified corn-soybean meal diets with 3,200 kcal of ME/kg would probably approximately 0.72% of the diet. A DL-Met vs L-cystine supplementation assay suggested that digestible cystine can supply no more than 52% of the total requirement for digestible SAA of chicks during the 3- to 6-wk growth period.