Regulation of global and specific mRNA translation by amino acids. (41/363)

A continuous supply of a complete complement of essential amino acids is a prerequisite for maintenance of optimal rates of protein synthesis in both liver and skeletal muscle. Deprivation of even a single essential amino acid causes a decrease in the synthesis of essentially all cellular proteins through an inhibition of the initiation phase of mRNA translation. However, the synthesis of all proteins is not repressed equally. Specific subsets of proteins, in particular those encoded by mRNAs containing a 5'-terminal oligopyrimidine (TOP) motif, are affected to a much greater extent than most proteins. The specific decrease in TOP mRNA translation is a result of an inhibition of the ribosomal protein S6 kinase, S6K1, and a concomitant decline in S6 phosphorylation. Interestingly, many TOP mRNAs encode proteins involved in mRNA translation, such as elongation factors eEF1A and eEF2, as well as the ribosomal proteins. Thus, deprivation of essential amino acids not only directly and rapidly represses global mRNA translation, but also potentially results in a reduction in the capacity to synthesize protein.  (+info)

Oral and intravenous tracer protocols of the indicator amino acid oxidation method provide the same estimate of the lysine requirement in healthy men. (42/363)

To investigate whether splanchnic uptake of the indicator amino acid ([1-(13)C] phenylalanine) during the fed state alters the estimate of lysine requirement, five healthy men were studied at graded levels of lysine intake, with either an oral or intravenous (IV) tracer protocol, in a randomized, crossover design. Splanchnic extraction of the oral tracer was expressed as the difference between the ratio of the enrichments in urinary phenylalanine between tracer protocols. The rate of release of (13)CO(2) from (13)C-phenylalanine oxidation (F(13)CO(2)) was measured and a two-phase linear regression crossover model was applied to determine the lysine requirement. Mean splanchnic extraction of the oral tracer was approximately 19%. Although actual F(13)CO(2) was higher during oral tracer infusion (P < 0.001), the breakpoint was not different from that determined with IV infusion (P = 0.98), with both yielding a mean lysine requirement of 36.6 mg/(kg. d). The upper 95% confidence intervals were 52.5 and 53.3 mg/(kg. d) for the oral and IV isotope infusions, respectively. These results demonstrate that routes of isotope administration using the indicator amino acid oxidation technique do not affect the estimated amino acid requirement. Therefore, the indicator amino acid oxidation method using the oral route, which is less invasive and allows for studies in vulnerable groups such as infants and children, should be the preferred method for studying amino acid requirements.  (+info)

Effect of folic acid and glycine supplementation on embryo development and folate metabolism during early pregnancy in pigs. (43/363)

The present work aimed to determine if different levels of prolificacy either by parity or by genetic origin are linked to folate metabolism. Nulliparous Yorkshire-Landrace (YL) and multiparous YL, and multiparous Meishan-Landrace (ML) sows were randomly assigned to two treatments: 0 ppm or 15 ppm folic acid+0.6% glycine. Supplements were given from the estrus before mating until slaughter on d 25 of gestation. At slaughter, embryo and endometrial tissues were collected to determine concentrations of DNA, protein, and homocysteine. Allantoic fluid samples were also collected to determine concentrations of folates, vitamin B12 and amino acids. Blood samples were taken at first estrus, at mating, and on d 8, 16, and 25 of gestation to determine serum concentrations of folates, vitamin B12, and relative total folate binding capacity (TFBC). Over the entire experiment, multiparous YL sows had higher average serum concentrations of folates than nulliparous YL sows (P < 0.05) but had similar serum concentrations of relative TFBC. Concentrations of folates and relative TFBC averaged higher in ML measured over the entire experiment than in multiparous YL sows (P < 0.05). Concentrations of serum vitamin B12 were higher in multiparous YL than in ML sows or YL nulliparous sows (P < 0.05) over the entire experiment. In allantoic fluid, folates, vitamin B12, and essential amino acids contents were significantly lower in ML than in YL multiparous sows (P < 0.05). The folic acid+glycine supplement increased concentrations of serum folates, but the increase was more marked in nulliparous YL sows (nulliparous x folic acid+glycine, P < 0.05). The folic acid+glycine supplement had no effect on litter size and embryo survival, but it tended to increase embryo DNA in multiparous YL sows (P = 0.06) but not in ML and nulliparous YL sows. Homocysteine was decreased by folic acid+glycine supplement in embryos from all sows, but in endometrium, the folic acid+glycine effect was dependent on parity (nulliparous x folic acid+glycine, P < 0.05). The effects of folic acid+glycine on litter size and embryo development and survival and some aspects of folate metabolism suggest that the basal dietary content of folic acid+glycine was adequate for ML and nulliparous YL sows but not to optimize embryo development in YL multiparous sows.  (+info)

Acute IGF-I infusion stimulates protein synthesis in skeletal muscle and other tissues of neonatal pigs. (44/363)

Studies have shown that protein synthesis in skeletal muscle of neonatal pigs is uniquely sensitive to a physiological rise in both insulin and amino acids. Protein synthesis in cardiac muscle, skin, and spleen is responsive to insulin but not amino acid stimulation, whereas in the liver, protein synthesis responds to amino acids but not insulin. To determine the response of protein synthesis to insulin-like growth factor I (IGF-I) in this model, overnight-fasted 7- and 26-day-old pigs were infused with IGF-I (0, 20, or 50 microg. kg(-1). h(-1)) to achieve levels within the physiological range, while amino acids and glucose were clamped at fasting levels. Because IGF-I infusion lowers circulating insulin levels, an additional group of high-dose IGF-I-infused pigs was also provided replacement insulin (10 ng. kg(-0.66). min(-1)). Tissue protein synthesis was measured using a flooding dose of L-[4-(3)H]phenylalanine. In 7-day-old pigs, low-dose IGF-I increased protein synthesis by 25-60% in various skeletal muscles as well as in cardiac muscle (+38%), skin (+24%), and spleen (+32%). The higher dose of IGF-I elicited no further increase in protein synthesis above that found with the low IGF-I dose. Insulin replacement did not alter the response of protein synthesis to IGF-I in any tissue. The IGF-I-induced increases in tissue protein synthesis decreased with development. IGF-I infusion, with or without insulin replacement, had no effect on protein synthesis in liver, jejunum, pancreas, or kidney. Thus the magnitude, tissue specificity, and developmental change in the response of protein synthesis to acute physiological increases in plasma IGF-I are similar to those previously observed for insulin. This study provides in vivo data indicating that circulating IGF-I and insulin act on the same signaling components to stimulate protein synthesis and that this response is highly sensitive to stimulation in skeletal muscle of the neonate.  (+info)

Adaptive, or stationary-phase, mutagenesis, a component of bacterial differentiation in Bacillus subtilis. (45/363)

Adaptive (stationary-phase) mutagenesis occurs in the gram-positive bacterium Bacillus subtilis. Furthermore, taking advantage of B. subtilis as a paradigm for the study of prokaryotic differentiation and development, we have shown that this type of mutagenesis is subject to regulation involving at least two of the genes that are involved in the regulation of post-exponential phase prokaryotic differentiation, i.e., comA and comK. On the other hand, a functional RecA protein was not required for this type of mutagenesis. The results seem to suggest that a small subpopulation(s) of the culture is involved in adaptive mutagenesis and that this subpopulation(s) is hypermutable. The existence of such a hypermutable subpopulation(s) raises important considerations with respect to evolution, the development of specific mutations, the nature of bacterial populations, and the level of communication among bacteria in an ecological niche.  (+info)

Acute response of net muscle protein balance reflects 24-h balance after exercise and amino acid ingestion. (46/363)

The purpose of this study was to determine if the acute anabolic muscle response to resistance exercise and essential amino acids (EAA) reflects the response over 24 h. Seven subjects participated in the following two 24-h studies: 1) resting (REST) and 2) rest plus resistance exercise and consumption of EAA (ES). Net balance (NB) across the leg was determined for four amino acids. [(13)C(6)]phenylalanine was infused to determine mixed muscle fractional synthetic rate (FSR). Twenty-four-hour FSR was significantly greater for ES than for REST (P = 0.003). Exchange of phenylalanine across the leg was -194 +/- 74 (SE) mg for ES and -371 +/- 88 mg for REST (P = 0.07) over 24 h and 229 +/- 42 mg (ES) and 28 +/- 15 mg (REST; P < 0.01) over 3 h corresponding to exercise and EAA consumption for ES. The difference in phenylalanine exchange between REST and ES was not different for measurements over 24 and 3 h. Increases in NB during ES were primarily the result of increases in protein synthesis. Results for other amino acids were similar. The acute anabolic response of muscle to EAA intake and exercise is additive to the response at rest and thus reflects the 24-h response.  (+info)

Hypercortisolemia alters muscle protein anabolism following ingestion of essential amino acids. (47/363)

Debilitating injury is accompanied by hypercortisolemia, muscle wasting, and disruption of the normal anabolic response to food. We sought to determine whether acute hypercortisolemia alters muscle protein metabolism following ingestion of a potent anabolic stimulus: essential amino acids (EAA). A 27-h infusion (80 microg. kg(-1). h(-1)) of hydrocortisone sodium succinate mimicked cortisol (C) levels accompanying severe injury (>30 microg/dl), (C + AA; n = 6). The control group (AA) received intravenous saline (n = 6). Femoral arteriovenous blood samples and muscle biopsies were obtained during a primed (2.0 micromol/kg) constant infusion (0.05 micromol. kg(-1). min(-1)) of l-[ring-(2)H(5)]phenylalanine before and after ingestion of 15 g of EAA. Hypercortisolemia [36.5 +/- 2.1 (C + AA) vs. 9.0 +/- 1.0 microg/dl (AA)] increased postabsorptive arterial, venous, and muscle intracellular phenylalanine concentrations. Hypercortisolemia also increased postabsorptive and post-EAA insulin concentrations. Net protein balance was blunted (40% lower) following EAA ingestion but remained positive for a greater period of time (60 vs. 180 min) in the C + AA group. Thus, although differences in protein metabolism were evident, EAA ingestion improved muscle protein anabolism during acute hypercortisolemia and may help minimize muscle loss following debilitating injury.  (+info)

Effect of soybean meal particle size on amino acid and energy digestibility in grower-finisher swine. (48/363)

A study was conducted using the ileal digestibility technique with grower-finisher pigs to evaluate the effects of particle size reduction of soybean meal (SBM) on amino acid and energy digestibility. Soybean meal was processed through a hammer mill to achieve average particle sizes of 900, 600, 300, and 150 microm. The treatments included the use of two soybean meal sources and soy protein concentrate. One source of SBM was ground to four different particle sizes (i.e., 949, 600, 389, 185); a second source was a common SBM source used in other trials (i.e., 800 microm). The soy protein concentrate had an average particle size of 385 microm. A low-protein (5% casein) diet was fed to determine endogenous amino acid losses. This experiment was conducted in a 7 x 7 Latin Square design in two replicates using 14 crossbred barrows ([Landrace x Yorkshire] x Duroc) that averaged 28 kg BW and 60 d of age. Animals were surgically fitted with a T-cannula at the distal ileum. Treatment diets were fed in meal form, initially at 0.09 kg BW(0.75) and at graded increases at each subsequent period. Pigs within replicate were fed a constant quantity of their treatment diet for a 5-d adjustment period followed by a 2-d collection of ileal digesta samples. Apparent and true digestibility of amino acids was calculated by use of chromic oxide (0.5%) as an indigestible marker. Apparent digestibility of isoleucine, methionine, phenylalanine, and valine increased linearly (P < 0.05) as particle size decreased. True digestibility of isoleucine, methionine, phenylalanine, and valine increased linearly (P < 0.05) as particle size decreased. When the essential amino acids were averaged, apparent digestibility increased (P < 0.10) from 83.5% to 84.9% as particle size decreased, whereas, nonessential amino acid digestibility increased only slightly (P > 0.15). Essential amino acid true digestibility increased numerically from 91.0% to 92.4% as particle size decreased. Energy digestibility was not affected by particle size (P > 0.15). These results suggest that a reduction in particle size of soybean meal resulted in a small increase in the digestibility of its amino acids with the essential amino acids being affected more than the nonessential amino acids. The largest improvement in digestibility, however, was obtained when the particle size was reduced to 600 microm.  (+info)