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Nitric oxide (NO) plays a central role as a cellular signaling molecule in health and disease. In the heart, NO decreases the rate of spontaneous beating and the velocity and extent of shortening and accelerates the velocity of relengthening. Since the cationic amino acid L-arginine (L-Arg) is the substrate for NO production by NO synthases (NOS), we tested whether the transporters that mediate L-Arg import in cardiac muscle cells represent an intervention point in the regulation of NO synthesis. Electrical currents activated by L-Arg with low apparent affinity in whole cell voltage-clamped rat cardiomyocytes were found to be rapidly and reversibly inhibited by NO donors. Radiotracer uptake studies performed on cardiac sarcolemmal vesicles revealed the presence of high-affinity/low-capacity and low-affinity/ high-capacity components of cationic amino acid transport that were inhibited by the NO donor S-nitroso-N-acetyl-DL-penicillamine. NO inhibited uptake in a noncompetitive manner with [K.sub.i] values of 275 and 827 nM for the high- and low-affinity component, respectively. Fluorescence spectroscopy experiments showed that millimolar concentrations of L-Arg initially promoted and then inhibited the release of endogenous NO in cardiomyocytes. Likewise, L-Arg currents measured in cardiac myocytes voltage clamped in the presence of 460 nM free intracellular [Ca.sup.2+], a condition in which a Ca-CaM complex should activate endogenous NO production, showed fast activation followed by inhibition of L-Arg transport. The NOS inhibitor N-nitro-L-arginine methyl ester, but not blockers of downstream reactions, specifically removed this inhibitory component. These results demonstrate that NO acutely regulates its own biosynthesis by modulating the availability of L-Arg via cationic amino acid transporters. lysine; amino acid transporters; nitric oxide synthase doi: 10.1152/ajpcell.00077.2010.

Mammalian cells respond to various kinds of stress, including nutritional stress, via pathways that are initiated by phosphorylation of the [alpha] subunit of the eukaryotic initiation factor 2 complex (elF2[alpha]). Because the models used to study elF2[alpha]-kinase--mediated responses to amino acid deficiency have commonly used media or diets devoid of 1 or more essential amino acids, we asked whether el F2[alpha]-kinase--mediated responses would be induced in animals fed a more typical diet that was not as imbalanced as one in which 1 essential amino acid is totally absent. To answer this question, we fed rats soy protein-based diets that were either adequate or limiting in sulfur-containing amino acids (SAA). Rats fed a SAA-deficient diet (3.4 g methionine equivalents/kg diet) grew more slowly than rats fed the control diet (5.86 g methionine equivalents/kg diet). Analysis of liver from rats fed these diets for 7 d showed that the SAA-deficient rats had higher levels of elF2[alpha] phosphorylation and higher levels of activating transcription factor (ATF) 4, ATF3, asparagine synthetase, solute carrier 7A11, cysteinyl-tRNA synthetase, and cystathionine [gamma]-lyase. On the other hand, components of the integrated stress response (ISR) known to promote apoptosis or translational recovery were not induced. Taken together, our results indicate that rats fed the SAA-deficient diet had a prolonged activation of an elF2[alpha] kinase that leads to upregulation of adaptive components of the ISR. J. Nutr. 140: 1080-1085, 2010. doi:10.3945/jn.109.120428.

We reporta unique mutation in the D-amino acid oxidase gene (R199W DAO) associated with classical adult onset familial amyotrophic lateral sclerosis (FALS) in a three generational FALS kindred, after candidate gene screening in a 14.52 cM region on chromosome 12q2223 linked to disease. Neuronal cell lines expressing R199W DAO showed decreased viability and increased ubiquitinated aggregates compared with cells expressing the wild-type protein. Similarly, lentiviral-mediated expression of R199W DAO in primary motor neuron cultures caused increased TUNEL labeling. This effect was also observed when motor neurons were cocultured on transduced astrocytes expressing R199W, indicating that the motor neuron cell death induced by this mutation is mediated by both cell autonomous and noncell autonomous processes. DAO controls the level of D-serine, which accumulates in the spinal coral in cases of sporadic ALS and in a mouse model of ALS, indicating that this abnormality may represent a fundamental component of ALS pathogenesis. motor neuron disease | neurodegeneration | linkage analysis | ubiquitinated aggregates | apoptosis doi/ 10.1073/pnas.0914128107

The purpose of this study was to investigate whether short-term amino acid supplementation could maintain a short-term net anabolic hormonal profile and decrease muscle cell damage during a period of high-intensity resistance training (overreaching), thereby enhancing recovery and decreasing the risk of injury and illness. Eight previously resistance trained males were randomly assigned to either a high branched chain amino acids (BCAA) or placebo group. Subjects consumed the supplement for 3 weeks before commencing a fourth week of supplementation with concomitant high-intensity total-body resistance training (overreaching) (3 x 6-8 repetitions maximum, 8 exercises). Blood was drawn prior to and after supplementation, then again after 2 and 4 days of training. Serum was analyzed for testosterone, cortisol, and creatine kinase. Serum testosterone levels were significantly higher (p KEY WORDS overreaching, testosterone, cortisol, muscle damage, BCAA

Dietary restriction extends healthy lifespan in diverse organisms and reduces fecundity (1,2). It is widely assumed to induce adaptive reallocation of nutrients from reproduction to somatic maintenance, aiding survival of food shortages in nature (3-6). If this were the case, long life under dietary restriction and high fecundity under full feeding would be mutually exclusive, through competition for the same limiting nutrients. Here we report a test of this idea in which we identified the nutrients producing the responses of lifespan and fecundity to dietary restriction in Drosophila. Adding essential amino acids to the dietary restriction condition increased fecundity and decreased lifespan, similar to the effects of full feeding, with other nutrients having little or no effect. However, methionine alone was necessary and sufficient to increase fecundity as much as did full feeding, but without reducing lifespan. Reallocation of nutrients therefore does not explain the responses to dietary restriction. Lifespan was decreased by the addition of amino acids, with an interaction between methionine and other essential amino acids having a key role. Hence, an imbalance in dietary amino acids away from the ratio optimal for reproduction shortens lifespan during full feeding and limits fecundity during dietary restriction. Reduced activity of the insulin/insulin-like growth factor signalling pathway extends lifespan in diverse organisms (7), and we find that it also protects against the shortening of lifespan with full feeding. In other organisms, including mammals, it may be possible to obtain the benefits to lifespan of dietary restriction without incurring a reduction in fecundity, through a suitable balance of nutrients in the diet., Dietary restriction, whereby food intake is reduced without leading to malnutrition, extends lifespan in many organisms: yeast (8), invertebrates (9) and mammals (1), including primates (10). In rodents and primates, [...]

Feline infectious peritonitis (FIP) is an uncommon, fatal, progressive, and immune-augmented disease of cats caused by feline coronavirus (FCoV) infection. Although FCoV is common in most domestic, feral, and nondomestic [...]

Intestinal nutrient transport is altered in response to changes in dietary conditions and luminal substrate level. It is not clear, however, whether an amino acid in the intestinal lumen can acutely affect its own absorption from a distant site. Our aim is to study the effect of an amino acid present in rat small intestinal segment on its own absorption from a proximal or distal site and elucidate the underlying mechanisms. The effect of instillation of alanine (Ala) in either jejunum or ileum on its own absorption at ileal or jejunal level was examined in vivo. The modulation of this intestinal regulatory loop by the following interventions was studied: tetrodotoxin (TTX) added to Ala, subdiaphragmatic vagotomy, chemical ablation of capsaicin-sensitive primary afferent (CSPA) fibers, and IV administration of calcitonin gene-related peptide (CGRP) antagonist. In addition, the kinetics of jejunal Ala absorption and the importance of [Na.sup.+]-dependent transport were studied in vitro after instilling Ala in the ileum. Basal jejunal Ala absorption [0.198 [+ or -] 0.018 [micro]mol x [cm.sup.-1] x 20 [min.sup.-1] (means [+ or -] SD)] was significantly decreased with the instillation of 20 mM Ala in the ileum or in an adjacent distal jejunal segment (0.12 [+ or -] 0.015; P < 0.0001 and 0.138 [+ or -] 0.014; P < 0.002, respectively). Comparable inhibition was observed in the presence of proline in the ileum. Moreover, basal Ala absorption from the ileum (0.169 [+ or -] 0.025) was significantly decreased by the presence of 20 mM Ala in the jejunum (0.103 [+ or -] 0.027; P < 0.01). The inhibitory effect on jejunal Ala absorption was abolished by TTX, subdiaphragmatic vagotomy, neonatal capsaicin treatment, and CGRP antagonism. In vitro studies showed that Ala in the ileum affects [Na.sup.+]-mediated transport and increases [K.sub.m] without affecting [V.sub.max]. Intraluminal amino acids control their own absorption from a distant part of the intestine, by affecting the affinity of the [Na.sup.+]-mediated Ala transporter, through a neuronal mechanism that involves CSPA and CGRP. control of amino acid absorption; intestinointestinal reflex; enteric nervous system

Idd3 is one of many gene regions that affect the development of type 1 diabetes (T1D) in the nonobese diabetic (NOD) mouse. Idd3 has been localized to a 650-kb region on chromosome 3 containing the IL-2 gene. Exon 1 of the IL-2 gene is polymorphic between the susceptible NOD and the protective C57BL/6 (B6) alleles, causing multiple amino acid changes that have been proposed to be responsible for the differing glycosylation status. To address whether this coding polymorphism recapitulates the disease suppression mediated by the B6 Idd3 allele, we generated knockin mice in which exon 1 of the B6 IL-2 allele replaces the homologous region in the NOD allele. We generated these mice by targeting the NOD allele of NOD/129 [F.sub.1] ES cells. IL-2 protein from the knockin mice showed the glycosylation pattern of the B6 IL-2 isoform, confirming that the amino acid differences encoded within exon 1 affect the glycosylation of the IL-2 protein. However, unlike NOD.B6 Idd3 congenic mice, the knockin mice were not protected from T1D. Furthermore, the difference in amino acid sequence in the IL-2 protein did not affect the level of expression of IL-2. This approach provides a general method for the determination of a functional role of a given genomic sequence in a disease process. Further, our result demonstrates that the variants in exon 1 of the IL-2 gene are not responsible for T1D suppression in NOD.B6 Idd3 mice, thereby supporting the hypothesis that variants in the regulatory region affecting expression levels are causative. Idd3 | NOD ES cells | flanking gene problem

Although amino acids (AA) synthesized by enteric microbiota in the upper gut of nonruminants can be absorbed, they do not necessarily make a net contribution to the host's AA supply. That depends on whether protein or nonprotein nitrogen sources are used for microbial protein production. We determined the contributions of urea, endogenous protein (EP), and dietary protein (DP) to microbial valine (MVAL) at the distal ileum of growing pigs, based on isotope dilutions after a 4-d continuous infusion of L-[1-[sup.13]C]valine to label EP and of [[sup.15]N [sup.15]N]urea. Eight barrows were assigned to either a cornstarch and soybean meal-based diet with or without 12% added fermentable fiber from pectin. Dietary pectin did not affect (P > 0.10) the contributions of the endogenous and DP to M.VAL. More than 92% of valine in microbial protein in the upper gut was derived from preformed AA from endogenous and DP, suggesting that de novo synthesis makes only a small contribution to microbial AA.

To reduce the impact of animal production on the environment, the CP content of the diet can be reduced by limiting the excess supply of AA. Improving the balance between AA relative to the requirement of the animal implies that we need to have accurate knowledge of the requirement of individual AA. The purpose of this study was to determine the Val requirement in postweaned piglets (12 to 25 kg) because Val is considered to be potentially limiting to performance after Lys, Met (and Cys), Thr, and Trp. The first experiment was carried out to identify a diet limiting in Lys supply. In this experiment, piglets were offered 1 of 3 diets: a low-CP diet containing low or adequate Lys concentrations [providing 1.0 and 1.2% standardized ileal digestible (SID) Lys, respectively] or a normal-CP diet with 1.2% SID Lys. Average daily gain of piglets receiving the diet containing 1.0% SID Lys was significantly less than that of piglets receiving diets containing 1.2% SID Lys at low or normal CP (486 vs. 522 g/d, respectively; P < 0.01). In Exp. 2, four diets with 1.0% SID Lys were used in a 2 x 2 factorial design, in which diets contained 57 or 70% SID Val:Lys in combination with 50 or 60% SID Ile:Lys. Independent of the Ile supply, feed intake and daily BW gain were, respectively, 15 and 20% less in piglets receiving diets providing 57% SID Val:Lys compared with piglets receiving 70% SID Val:Lys (P < 0.001). The Ile content of the diet did not affect feed intake or daily BW gain (P > 0.10). Experiment 3 was conducted to evaluate the response of piglets to an increasing Val supply provided by 2 sources of L-Val differing in the degree of purity. Increasing the Val supply from 58 to 66% SID Val:Lys resulted in a linear increase in both feed intake and daily gain by 24 and 30%, respectively (P < 0.001). No difference was observed between both sources of L-Val (P > 0.10). Experiment 4 was a dose-response study using 5 concentrations of Val supply (ranging from 60 to 80% SID Val:Lys). The estimated SID Val:Lys requirements for maximizing ADFI, ADG, and G:F were, respectively, 74, 70, and 68% using a linear-plateau model, and 81, 75, and 72% using a curvilinear-plateau model. Plasma Val, plasma [alpha]-ketoisovaleric acid, Ile, and Leu concentrations after an overnight fast increased with increasing Val supply (P < 0.001). The results of these experiments indicated that the SID Val:Lys was at least 70%, which was slightly greater than the current NRC recommendation. Key words: branched-chain amino acid, feed intake, growth, pig, requirement, valine

Delayed allograft function (DGF) is a common adverse event in postrenal transplantation. The etiology of DGF is thought to include both nonimmunologic (donor age, cold ischemia time, and recipient race) and immunologic factors. We examined the association of DGF with amino acid mismatches at 66 variable sites of the HLA-A molecule in a prospective cohort study of 697 renal transplant recipients of deceased donors. Using a multivariate logistic regression model adjusted for nonimmunologic risk factors, we show that combinations of a few amino acid mismatches at crucial sites of HLA-A molecules were associated with DGF. In Caucasian recipients, a mismatch at position 62, 95, or 163, all known to be functionally important within the antigen recognition site, was associated with an increased risk for DGF. Furthermore, a decreased risk for DGF was associated with a mismatch at HLA-A family-specific sites (149, 184, 193, or 246), indicating that evolutionary features of HLA-A polymorphism separating HLA-A families and lineages among donor-recipient pairs may correlate with the magnitude of alloreactivity influencing the development of DGF. These findings suggest that amino acid polymorphisms at functionally important positions at the antigen recognition site of the HLA-A molecule have a significant influence on DGF. HLA alleles | kidney transplantation | HLA mismatches | MHC

Ruminally cannulated Rambouillet wether lambs were used in three 6 x 6 Latin square experiments (n = 6/experiment) to determine which essential AA limit N retention. Lambs (BW = 36.9 [+ or -] 1.9 kg for Exp. 1, 35.1 [+ or -] 1.4 kg for Exp. 2, and 46.0 [+ or -] 1.3 kg for Exp. 3) were housed in metabolism crates and limit-fed (DMI = approx. 1.8% of BW daily) twice daily a soybean hull-based diet low in ruminally undegradable protein. Treatments for Exp. 1 were continuous abomasal infusions of a solution (500 mL/d) containing 1) no AA (CON), 2) a mixture of 10 essential AA and 2 nonessential AA (10EAA), 3) 10EAA with Met removed, 4) 10EAA with Lys removed, 5) 10EAA with His removed, and 6) 10EAA with Thr removed. Treatments for Exp. 2 were abomasal infusions of 1) CON, 2) 10EAA, 3) 10EAA with Leu, Ile, and Val removed (-BCAA), 4) 10EAA with Arg removed, 5) 10EAA with Phe removed, and 6) 10EAA with Trp removed. Treatments for Exp. 3 were abomasal infusions of 1) CON, 2) 10EAA, 3) -BCAA, 4) 10EAA with Leu removed, 5) 10EAA with Ile removed, and 6) 10EAA with Val removed. All lambs received continuous infusions of acetate and propionate into the rumen and dextrose into the abomasum to supply additional energy. Periods were 7 d: 3 d for adaptation to abomasally infused treatments and 4 d for fecal and urinary collections. Blood samples were collected 3 h after feeding on d 7. In all 3 experiments, N retention was greater (P < 0.10) for lambs receiving 10EAA vs. CON, demonstrating that the basal AA supply from CON was limiting. Removal of each of the essential AA from 10EAA decreased (P < 0.10) their concentrations in plasma (except for Trp), indicating that 10EAA supplied these AA in excess of the animal's requirement. In Exp. 1, N retention (g/d) decreased (P < 0.10) in response to the removal of Met and Thr, but was not affected by removal of Lys and His from 10EAA. In Exp. 2, N retention decreased (P < 0.10) in response to removal of all 3 branched-chain AA, Arg, and Trp, whereas the removal of Phe from 10EAA did not affect N retention. In Exp. 3, N retention decreased (P < 0.10) in response to removal of branched-chain AA and Val, but was not affected by the omission of Leu and Ile from 10EAA. The results of this research demonstrated that Met, Thr, Arg, Trp, and Val limited N retention of lambs fed a diet low in ruminally undegradable protein. Key words: lamb, limiting amino acid, nitrogen retention

The existing data on the safe upper limits of amino acid intake in humans is essentially observational; how much do individuals ingest and what side effects do they have? There are numerous studies in humans comparing the effects of high doses of amino acids given as protein bound vs. as free amino acids. These studies have shown that protein-bound amino acids have much less effect on plasma levels of the test amino acid, because protein intake stimulates protein synthesis as another sink for the increased amino acid intake. In practice, the highest amino acid intakes occur with free amino acid supplements that may be ingested by athletes who believe that the amino acids will benefit them in training and/or performance. Previously, in a piglet study, we were able to define the point at which maximal phenylalanine oxidation occurred, above which plasma phenylalanine concentration and body balance rose exponentially. We regard this value of maximal disposal (oxidation) of an amino acid as one metabolic marker of the upper limit of intake. Recently, others have demonstrated a similar maximal oxidation rate for leucine in rats. Based on these experimental data and the paucity of published human data in controlled experiments, we think that a systematic approach needs to be undertaken to define the maximal oxidation rate for all dietary indispensable amino acids and other amino acids that may be ingested in excess by humans. We believe that this will provide a rational basis to begin to define the upper limits of tolerance for dietary amino acids. However, some amino acids, such as threonine and methionine, will be more difficult to study, because they have more than 1 route of disposal or very complex metabolic regulation, in which case defining their upper limits will be more multifaceted.

Tumstatin is an angiogenesis inhibitor that binds to [alpha]v[beta]3 integrin and suppresses tumor growth. Previous deletion mutagenesis studies identified a 25-aa fragment of tumstatin (tumstatin peptide) with in vitro antiangiogenic activity. Here, we demonstrate that systemic administration of this tumstatin peptide inhibits tumor growth and angiogenesis. Site-directed mutagenesis identified amino acids L, V, and D as essential for the antiangiogenic activity of tumstatin. The tumstatin peptide binds to [alpha]v[beta]3 integrin on proliferating endothelial ceils and localizes to select tumor endothelium in vivo. Using 3D molecular modeling, we identify a putative interaction interface for tumstatin peptide on [alpha]v[beta]3 integrin. The antitumor activity of the tumstatin peptide, in combination with bevacizumab (anti-VEGF antibody), displays significant improvement in efficacy against human renal cell carcinoma xenografts when compared with the single-agent use. Collectively, our results demonstrate that tumstatin peptide binds specifically to the tumor endothelium, and its antiangiogenic action is mediated by [alpha]v[beta]3 integrin, and, in combination with an anti-VEGF antibody it exhibits enhanced tumor suppression of renal cell carcinoma. angiogenesis | [alpha]v[beta]3 integrin | bevacizumab | tumstatin peptide

A study was conducted to evaluate the extent and dynamics of whole body protein deposition (Pd) and changes in chemical and physical body composition after a period of AA intake restriction in entire male pigs with high lean-tissue growth potentials. Fifty-eight entire male pigs (initial BW 15.8 [+ or -] 0.9 kg) were allotted to 1 of 3 dietary AA levels between 15 and 38 kg of BW: control (15% above requirements), AA-15% (15% below requirements), and AA-30% (30% below requirements). Thereafter, pigs were fed diets not limiting in AA content. Throughout the experiment, pigs were scale-fed at 90% of estimated voluntary daily DE intake. Representative pigs were slaughtered at 15, 38, 53, 68, or 110 kg of BW to monitor changes in body composition. Between 15 and 38 kg of BW, restriction of AA intake reduced BW gain (P < 0.01; 794, 666, and 648 g/d for control, AA-15%, and AA-30%, respectively). At 38 kg of BW, AA intake restriction increased whole body lipid (LB) content (P < 0.01; 11.3, 14.3, 17.5% of empty BW), and the LB-to-whole body protein (PB) ratio (LB/PB; P < 0.02; 0.68, 0.88, 1.10 for control, AA-15%, and AA-30%, respectively). Relationships between PB versus whole body water and PB versus whole body ash were not affected by dietary treatments (P > 0.10). At 110 kg of BW and based on BW, PB, and LB/PB, complete compensatory growth (CG) was achieved. Body weight gain between 38 and 110 kg of BW was inversely related to previous dietary AA levels (P < 0.01; 1,089, 1,171, and 1,185 g/d for control, AA-15%, and AA-30%, respectively). For pigs on the control diet, and based on N-balance data, Pd increased with BW, from 172 g/d at 40 kg of BW to 226 g/d at 82 kg of BW. At 40 kg of BW, Pd was greater (P < 0.05) for pigs on the AA-15% (205 g/d) and AA-30% (191 g/d) diets than pigs on the control diet (172 g/d). These findings indicate that pigs with high lean-tissue growth potentials are more likely to express compensatory Pd and their genetically determined upper limit to Pd (PdMax) after a period of AA intake restriction. This study confirms previous findings that BW effects on PdMax are small in growing pigs between 40 and 80 kg of BW. It is suggested that CG and compensatory Pd after a period of AA intake restriction is constrained by the pig's PdMax and is driven by a target LB/PB. Combined with previous observations in our laboratory, these results suggest that CG after a period of AA intake restriction tends to occur only when pigs are within the energy-dependent phase of lean-tissue growth and not when the genetically determined upper limit to lean-tissue growth, or PdMax, determines growth performance. Key words: amino acid intake, body composition, compensatory growth, pig

A study was conducted to evaluate the extent and dynamics of whole body protein deposition and changes in chemical and physical body composition after a period of AA intake restriction in growing barrows with medium lean tissue growth potentials. Forty Yorkshire barrows (initial BW 14.4 [+ or -] 1.6 kg) were scale-fed at 75% of estimated voluntary daily DE intake up to 35 kg of BW and assigned to 1 of 2 diets: AA adequate (AA+; 20% above requirements; NRC, 1998) and AA deficient (AA-; 40% below requirements; restriction phase). Thereafter (re-alimentation phase), pigs from both dietary AA levels were scale-fed or fed ad libitum diets that were not limiting in AA. Body weight gain and body composition, based on serial slaughter, were monitored during the 34-d re-alimentation phase. During the restriction phase AA intake restriction reduced BW gains (556 vs. 410 g/d; P < 0.001; AA+ and AA-, respectively). At 35 kg of BW, AA intake restriction increased whole body lipid content (11.1 vs. 17.5% of empty BW; P < 0.05) and the whole body lipid to body protein ratio (0.65 vs. 1.20; P < 0.01) and reduced body protein content (17.1 vs. 14.6% of empty BW; P < 0.01) and body water content (68.2 vs. 63.9%; P < 0.05). The relationships between body protein vs. body water and body protein vs. body ash content were not altered by previous AA intake restriction or by feeding level during the re-alimentation phase (P > 0.10). Throughout the re-alimentation phase, there were no interactive effects of time, feeding level, and previous AA intake level on growth performance, body protein, and body lipid content (P > 0.10). During the re-alimentation phase, body protein deposition, derived from the linear regression analysis of body protein content vs. time, was not affected by feeding level and previous AA intake level (P > 0.10; 156 g/d for AA- vs. 157 g/d for AA+). Based on BW and body protein content, it can be concluded that no compensatory body protein deposition occurred in barrows, with medium lean tissue growth potential after AA intake restriction between 15 and 35 kg of BW. It is suggested that the upper limit to body protein deposition was the main factor that limited the extent of compensatory body protein deposition in this population of pigs. The concept of an upper limit to body protein deposition may be used to explain why compensatory growth is observed in some studies and not in others. Key words: amino acid intake, body composition, compensatory growth, pig

Over the last 50 yr, the study of intestinal peptide transport has rapidly evolved into a field with exciting nutritional and biomedical applications. In this review, we describe from a historical and current perspective intestinal peptide transport, the importance of peptides to whole-body nutrition, and the cloning and characterization of the intestinal peptide transporter, PepT1. We focus on the nutritional significance of peptide transport and relate these findings to livestock and poultry. Amino acids are transported into the enterocyte as free AA by a variety of AA transporters that vary in substrate specificity or as di- and tripeptides by the peptide transporter, PepT1. Expression of PepT1 is largely restricted to the small intestine in most species; however, in ruminants, peptide transport and activity is observed in the rumen and omasum. The extent to which peptides are absorbed and utilized is still unclear. In ruminants, peptides make a contribution to the portal-drained visceral flux of total AA and are detected in circulating plasma. Peptides can be utilized by the mammary gland for milk protein synthesis and by a variety of other tissues. We discuss the factors known to regulate expression of PepT1 including development, diet, hormones, diurnal rhythm, and disease. Expression of PepT1 is detected during embryological stages in both birds and mammals and increases with age, a strategic event that allows for the immediate uptake of nutrients after hatch or birth. Both increasing levels of protein in the diet and dietary protein deficiencies are found to upregulate the peptide transporter. We also include in this review a discussion of the use of dietary peptides and potential alternate routes of nutrient delivery to the cell. Our goal is to impart to the reader the nutritional implications of peptide transport and dietary peptides and share discoveries that shed light on various biological processes, including rapid establishment of intestinal function in early neonates and maintenance of intestinal function during fasting, starvation, and disease states. Key words: amino acid absorption, dietary regulation, intestine, PepT1, peptide, transporter

Human [gamma][delta]T lymphocytes are a subset of T cells and are a first line of defense against microbes and tumors. These [gamma][delta] T cells can be primed by nitrogen-containing bisphosphonates, and certain short-chain alkylamines. These primed [gamma][delta] T cells have an enhanced capacity to proliferate and to secrete cytokines upon ex vivo exposure to a wide variety of microbes and tumor cells. The largest dietary source of alkylamines is L-theanine, an amino acid unique to tea beverages that is catabolized to ethylamine. Supplementation of subjects with capsules containing L-theanine and catechins has recently been shown to decrease the incidence of cold and flu symptoms, while enhancing [gamma][delta] T cell function.

We determined complete genome sequences of lineage 2 West Nile virus (WNV) strains isolated from patients in South Africa who had mild or severe WNV infections. These strains had previously [...]

: Introduction: Failure of protease‐inhibitor (PI)‐based second‐line antiretroviral therapy (ART) with medication adherence but no protease drug resistance mutations (DRMs) is not well understood. This study investigated the involvement of gp41 and gag as alternative mechanisms, not captured by conventional resistance testing and particularly relevant in resource‐limited settings where third‐line ART is limited. Methods: We evaluated gp41 and gag for unique amino acids in seven subtype A infected Kenyans failing second‐line therapy with no PI resistance yet detectable lopinavir (query dataset), compared to seven similar‐setting patients with PI resistance or undetectable lopinavir and 69 publically available subtype A Kenyan whole‐genomes sequences. Results: Three gp41 (607T, 641L, 721I) and four gag (124S, 143V, 339P, 357S) amino acids were significantly more frequent in the query dataset compared to the other datasets, with significantly high co‐occurrence. Conclusion: The genotypic analysis of a unique group of HIV‐1 subtype A infected patients, identified seven amino acids that could potentially contribute to a multi‐gene mechanism of PI‐based ART failure in the absence of PI DR mutations., Introduction The main cause of HIV‐1 antiretroviral therapy (ART) failure is drug resistance (DR) development. In resource‐limited settings (RLS) there are only two recommended ART lines: a non‐nucleoside reverse transcriptase [...]

Background/Objectives: Recent metabolomics technique reveals a plasma-free amino acid (PFAA)-based metabolite signature that is suggestive of altered PFAAs being an early manifestation of obesity-related insulin resistance. However, the PFAA profiles within non-obese, but more insulin-resistant Asians are not well researched. Compared with Caucasians, Asian populations have more central adiposity, which is generally regarded as metabolically more adverse, but the underlying mechanisms remain unclear. In the present study, we examined whether PFAA profiling was at least one important factor mediating central adiposity and insulin resistance, and aid in cardiovascular risk assessment in healthy Asians with normal body weight. Subject/Methods: This was a cross-sectional study. A total of 190 healthy men (n=87 with a mean[+ or -]s.d. body mass index (BMI) of 23.5[+ or -]3.5 kg/m[sup.2]) and women (n=103 with a mean[+ or -]s.d. BMI of 21.4[+ or -]3.7 kg/m[sup.2]) residing in Singapore took part in this study. PFAA levels were measured by using an amino acid analyzer. Basic anthropometric measurements, fasting blood glucose, fasting serum insulin and lipid profiles were obtained using standard protocols. Results: Seven out of 18 amino acids were significantly correlated with measures of obesity (for example, waist circumference; waist-to-hip ratio and BMI) in current participants. Among them, the plasma concentrations of five amino acids, including Phe, Tyr, Met, Ala and His were positively associated with waist-to-hip ratio. With the exception of His, which had no association with insulin resistance, Phe, Tyr, Met and Ala were significantly associated with hyperinsulinemia and insulin resistance (P [less than] 0.05). In contrast, no associations were observed between circulating BCAAs (that is, Val, Leu, Ile), measures of obesity and insulin resistance. However, significant inverse associations were observed between BCAAs and total cholesterol and high-density lipoprotein. Conclusions: We found that central adiposity was associated with alterations of specific amino acids. As a result, PFAAs may serve as metabolite predictors of hyperglycemia, hyperinsulinemia and dyslipidemia in healthy participants. European Journal of Clinical Nutrition (2017) 71, 1080-1087; doi: 10.1038/ejcn.2017.34; published online 5 April 2017, Author(s): X Bi [1]; S L Tey [1]; Y T Loo [1]; C J Henry [1, 2] Introduction Diabetes mellitus (DM) is an increasingly serious public health concern globally, which [...]

Insulin and amino acids act independently to stimulate protein synthesis in skeletal muscle of neonatal pigs, and the responses decrease with development. The purpose of this study was to compare the separate effects of fed levels of INS and AA on the activation of signaling components leading to translation initiation and how these responses change with development. Overnight-fasted 6- (n = 4/group) and 26-day-old (n = 6/ group) pigs were studied during 1) euinsulinemic-euglycemic-euaminoacidemic conditions (controls), 2) euinsulinemic-euglycemic-hyperaminoacidemic clamps (AA), and 3) hyperinsulinemic-euglycemic-euaminoacidemic clamps (INS). INS, but not AA, increased the phosphorylation of protein kinase B (PKB) and tuberous sclerosis 2 (TSC2). Both INS and AA increased protein synthesis and the phosphorylation of mammalian target of rapamycin (mTOR), ribosomal protein S6 kinase-1, and eukaryotic initiation factor (eIF)4E-binding protein 1 (4E-BP1), and these responses were higher in 6-day-old compared with 26-day-old pigs. Both INS and AA decreased the binding of 4E-BP1 to eIF4E and increased eIF4E binding to eIF4G; these effects were greater in 6-day-old than in 26-day-old pigs. Neither INS nor AA altered the composition of mTORC1 (raptor, mTOR, and G[beta]L) or mTORC2 (rictor, mTOR, and G[beta]L) complexes. Furthermore, neither INS, AA, nor age had any effect on the abundance of Rheb and the phosphorylation of AMP-activated protein kinase and eukaryotic elongation factor 2. Our results suggest that the activation by insulin and amino acids of signaling components leading to translation initiation is developmentally regulated and parallels the developmental decline in protein synthesis in skeletal muscle of neonatal pigs. neonates; mammalian target of rapamycin; tuberous sclerosis; raptor; protein synthesis

Schizophrenia is a chronic, complex and heterogeneous mental disorder, with pathological features of disrupted neuronal excitability and plasticity within limbic structures of the brain. These pathological features manifest behaviorally as positive symptoms (including hallucinations, delusions and thought disorder), negative symptoms (such as social withdrawal, apathy and emotional blunting) and other psychopathological symptoms (such as psychomotor retardation, lack of insight, poor attention and impulse control). Altered glutamate neurotransmission has for decades been linked to schizophrenia, but all commonly prescribed antipsychotics act on dopamine receptors. LY404039 is a selective agonist for metabotropic glutamate 2/3 (mGlu2/3) receptors and has shown antipsychotic potential in animal studies. With data from rodents, we provide new evidence that mGlu2/3 receptor agonists work by a distinct mechanism different from that of olanzapine. To clinically test this mechanism, an oral prodrug of LY404039 (LY2140023) was evaluated in schizophrenic patients with olanzapine as an active control in a randomized, three-armed, double-blind, placebo-controlled study. Treatment with LY2140023, like treatment with olanzapine, was safe and well-tolerated; treated patients showed statistically significant improvements in both positive and negative symptoms of schizophrenia compared to placebo (P [less than] 0.001 at week 4). Notably, patients treated with LY2140023 did not differ from placebo-treated patients with respect to prolactin elevation, extrapyramidal symptoms or weight gain. These data suggest that mGlu2/3 receptor agonists have antipsychotic properties and may provide a new alternative for the treatment of schizophrenia., Author(s): Sandeep T Patil [1, 13]; Lu Zhang [1]; Ferenc Martenyi (corresponding author) [2]; Stephen L Lowe [3]; Kimberley A Jackson [4]; Boris V Andreev [5]; Alla S Avedisova [6]; [...]

Aromatic amino acids in the brain function as precursors for the monoamine neurotransmitters serotonin (substrate tryptophan) and the catecholamines [dopamine, norepinephrine, epinephrine; substrate tyrosine (Tyr)]. Unlike almost all other neurotransmitter biosynthetic pathways, the rates of synthesis of serotonin and catecholamines in the brain are sensitive to local substrate concentrations, particularly in the ranges normally found in vivo. As a consequence, physiologic factors that influence brain pools of these amino acids, notably diet, influence their rates of conversion to neurotransmitter products, with functional consequences. This review focuses on Tyr and phenylalanine (Phe). Elevating brain Tyr concentrations stimulates catecholamine production, an effect exclusive to actively firing neurons. Increasing the amount of protein ingested, acutely (single meal) or chronically (intake over several days), raises brain Tyr concentrations and stimulates catecholamine synthesis. Phe, like Tyr, is a substrate for Tyr hydroxylase, the enzyme catalyzing the ratelimiting step in catecholamine synthesis. Tyr is the preferred substrate; consequently, unless Tyr concentrations are abnormally low, variations in Phe concentration do not affect catecholamine synthesis. Unlike Tyr, Phe does not demonstrate substrate inhibition. Hence, high concentrations of Phe do not inhibit catecholamine synthesis and probably are not responsible for the low production of catecholamines in subjects with phenylketonuria. Whereas neuronal catecholamine release varies directly with Tyr-induced changes in catecholamine synthesis, and brain functions linked pharmacologically to catecholamine neurons are predictably altered, the physiologic functions that utilize the link between Tyr supply and catecholamine synthesis/release are presently unknown. An attractive candidate is the passive monitoring of protein intake to influence protein-seeking behavior.

Liver failure is associated with hepatic encephalopathy (HE). An imbalance in plasma levels of aromatic amino acids (AAA) phenylalanine, tyrosine, and tryptophan and branched chain amino acids (BCAA) and their BCAA/AAA ratio has been suggested to play a causal role in HE by enhanced brain AAA uptake and subsequently disturbed neurotransmission. Until recently, data on this subject and the role of the liver and splanchnic bed were scarce, particularly in humans, due to inaccessibility of portal and hepatic veins. Here, we discuss, against a background of relevant literature, data obtained in patients undergoing liver resection or with a transjugular intrahepatic portasystemic stent shunt (TIPSS), where these veins are accessible. The BCAA/AAA ratio remained unchanged after major liver resection, but plasma AAA levels were inversely correlated (P < 0.001) with residual liver volume, in keeping with the observed hepatic AAA uptake. In patients with stable cirrhosis and a TIPSS, the plasma BCAA/AAA ratio was lower than in controls (1.19 [+ or -] 0.09 vs. controls: 3.63 [+ or -] 0.34). Gastrointestinal bleeding in cirrhotics with a TIPSS induced disturbances in BCAA levels and the BCAA/AAA ratio and induced catabolism, which could partly be corrected by isoleucine administration. AAA may be important in the pathogenesis of HE, but it is unlikely that they are the sole factors. HE most likely is a syndrome with multifactorial pathogenesis, where hyperammonemia, AAA/BCAA imbalances, inflammation, brain edema, and neurotransmitter changes interact. Novel therapies to normalize AAA levels in patients with liver failure (such as the molecular adsorbent recirculating system dialysis device) should probably be combined with supplementation of e.g. isoleucine and enhancing ammonia excretion by the kidneys.

Arginine (ARG) and its related amino acids (AAs) ornithine (ORN) and citrulline (CIT) find a range of applications as dietary supplements in subgroups of healthy subjects (e.g., bodybuilders) and patients with acute or chronic malnutrition. These AAs appear to be well utilized in humans with, in general, a rapid return of blood concentrations to basal values (i.e., within 5-8 h) and low absolute and relative excretion in urine (< 5% of administered dose). Based on published data for the maximum observed plasma concentrations ([C.sub.max]) after administration of doses in the range 5 to 10 g, CIT appeared to present relatively better absorption and systemic bioavailability than ARG and ORN. The few relevant dose-ranging studies available include 1 limited to a single subject receiving 5- to 20-g doses of ornithine [alpha]-ketoglutarate and another in which 8 subjects received from 5 to 15 g of CIT. Comparison of these 2 studies further indicates that CIT has higher bioavailability than ORN. The pharmacokinetics and metabolism of these AAs are modified by the coadministration of a salt such as [alpha]-ketoglutarate that modifies AA metabolism, as has clearly been demonstrated for ornithine [alpha]-ketoglutarate. Concomitant administration of a meal leads to a 15- to 30-min delay in [C.sub.max]. Finally, data from various pharmacokinetic studies together with basic physiology and biochemistry indicate that ARG is a net urea producer and ORN has a nitrogen-sparing effect, whereas CIT is neutral. However, most of the studies performed to date carry methodological weaknesses and are difficult to compare because of a number of confounding factors. To date, there have been no pharmacokinetic studies on the long-term administration of these AAs in healthy subjects despite the need to determine the safe upper limit of daily intake.

The calcium-sensing receptor (CAR) is recognized as a member of class 3 of the G-protein coupled receptor superfamily. Members of this subgroup, which have large N-terminal extracellular domains, include receptors that respond specifically to the amino acid glutamate; receptors that respond to the glutamate analogue, [gamma]-amino butyric acid; and several receptors that act as broad-spectrum amino acid sensors. The CaR is one of these broad-spectrum amino acid sensors that, along with several other members of the subgroup, also responds to extracellular [Ca.sup.2+]. In this mini-review, we consider evidence that the CaR is a sensor of aromatic amino acids, that it has broad-spectrum amino acid sensing properties, that it provides an amino acid binding site in its extracellular N-terminal Venus Fly Trap domain, and that amino acids have a physiological impact on systems in which the CaR is expressed.

Aquaporins (AQP) are a family of membrane channels primarily responsible for conducting water across cellular membranes. The availability of a large body of high resolution structural data along with numerous atomic-scale simulation studies have resulted in an unprecedented level of understanding of the mechanism of function and selectivity in AQP. In this article, after summarizing major highlights of structure-functional studies of AQP, we will report on some of our recent large-scale molecular dynamics simulations investigating the mechanisms of permeation of various substances through pure lipid bilayers and through multiple pathways provided by tetrameric structures of different AQP. Comparison of the results obtained for structurally highly homologous, but functionally distinct, AQP allowed us to identify novel mechanisms of gating and selectivity of these channels and to design mutants with experimentally verified, altered properties. When applicable, special attention will be given to specific aromatic amino acids and their involvement in various functional aspects of AQP.

Oral supplements of arginine and citrulline increase local nitric oxide (NO) production in the small intestine and this may be harmful under certain circumstances. Gastrointestinal toxicity was therefore reviewed with respect to the intestinal physiology of arginine, citrulline, ornithine, and cystine (which shares the same transporter) and the many clinical trials of supplements of the dibasic amino acids or N-acetylcysteine (NAC). The human intestinal dibasic amino acid transport system has high affinity and low capacity. L-Arginine (but not lysine, ornithine, or D-arginine) induces water and electrolyte secretion that is mediated by NO, which acts as an absorbagogue at low levels and as a secretagogue at high levels. The action of many laxatives is NQ mediated and there are reports of diarrhea following oral administration of arginine or ornithine. The clinical data cover a wide span of arginine intakes from 3 g/d to >100 g/d, but the standard of reporting adverse effects (e.g. nausea, vomiting, and diarrhea) was variable. Single doses of 3-6 g rarely provoked side effects and healthy athletes appeared to be more susceptible than diabetic patients to gastrointestinal symptoms at individual doses >9 g. This may relate to an effect of disease on gastrointestinal motility and pharmacokinetics. Most side effects of arginine and NAC occurred at single doses of >9 g in adults (>140 mg/kg) often when part of a daily regime of ->30 g/d (>174 mmol/d). In the case of arginine, this compares with the laxative threshold of the nonabsorbed disaccharide alcohol, lactitol (74 g or 194 mmol). Adverse effects seemed dependent on the dosage regime and disappeared if divided doses were ingested (unlike lactitol). Large single doses of poorly absorbed amino acids seem to provoke diarrhea. More research is needed to refine dosage strategies that reduce this phenomenon. It is suggested that dipeptide forms of arginine may meet this criterion.

Arginine is a nonessential amino acid in the normal physiological state that becomes conditionally essential during periods of hypermetabolic stress. Recent literature supports the hypothesis that arginine plays an important role in the intermediary metabolism of the critically ill patient. Current critical care literature is conflicting on arginine use in the clinical setting, with some proposing it as a panacea, whereas others report it as poison. Multiple individual reports and at least 5 major meta-analyses using combinations of immune-modulating nutrients have reported mostly beneficial results, but few have evaluated the effects of arginine when given as a single supplemental nutrient. This review attempts to objectively analyze the literature and evaluate the potential role of arginine in the critical care setting.

Lysine cannot be synthesized by mammals and, as a consequence, is an indispensable amino acid. The main role of lysine is to participate in protein synthesis. The catabolism of lysine is principally located in the liver. Lysine released from digested protein undergoes a significant first-pass metabolism of ~30 to 42% in humans and piglets. An important question regarding the biological basis of the requirement of lysine is the possible participation of microbial de novo synthesized amino acids in the whole-body fluxes. Recent intake recommendations to meet the tysine requirement range from 64 to 30 mg/(kg * d) for 0.5-y infants and adults (> 18 y), respectively. Lysine intake in the Western human diet is in the range 40-180 mg/(kg * d). An upper limit of 300-400 mg/(kg * d) can be considered in humans.

Differences in lysine and arginine requirements among various species such as omnivores (humans, pigs, rats, dogs), carnivores (cats), herbivores (rabbits, horses), ruminants (cattle), poultry, and fish, are covered in detail in this article. Although lysine is classified as an indispensable amino acid across species, the classification of arginine as either an indispensable or dispensable amino acid is more ambiguous because of differences among species in rates of de novo arginine synthesis. Because lysine is most often the limiting amino acid in the diet, its requirement has been extensively studied. By use of the ideal protein concept, the requirements of the other indispensable amino acids can be extrapolated from the lysine requirement. The successful use of this concept in pigs is compared with potential application of the ideal protein concept in humans. The current dietary arginine requirement varies widely among species, with ruminants, rabbits, and rats having relatively low requirements and carnivores, fish, and poultry having high requirements. Interspecies differences in metabolic arginine utilization and reasons for different rates of de novo arginine synthesis are reviewed in detail, as these are the primary determinants of the dietary arginine requirement. There is presently no dietary requirement for humans of any age, although this needs to be reassessed, particularly in neonates. A thorough understanding of the factors contributing to the lysine and arginine requirements in different species will be useful in our understanding of human amino acid requirements.

Citrulline (CIT) is an amino acid that is not involved in protein synthesis but that is tightly linked to arginine (ARG) metabolism. CIT displays a very specific metabolism: In the 1980s, Windmuller demonstrated that the small intestine releases CIT, which is mainly taken up by the kidney and metabolized into ARG. Because CIT is not taken up by the liver, this ARG-CIT-ARG cycle can be seen as a means of protecting dietary ARG from liver degradation and of sustaining protein homeostasis. These observations have led to the concept that plasma CIT concentration would be a good marker of intestinal failure in short bowel syndrome. Hence, in massive intestinal resection, citrullinemia is greatly reduced, and this is proportional to the severity of the intestinal disease. This concept was then extended to other situations in which the intestinal function is compromised. The data strongly suggest that CIT may be a conditionally essential amino acid in situations where the intestinal function is compromised. Recent data support this idea. Thus, CIT supplementation is able to restore nitrogen balance, generate large amounts of ARG in rats with short bowel syndrome, and increase muscle protein content (+20%) as well as muscle protein synthesis (+90%) in elderly malnourished rats. Finally, recent data indicate that CIT per se could be able to stimulate muscle protein synthesis. Hence, CIT could play a pivotal role in maintaining protein homeostasis, and the determination of the underlying mechanisms involved in its action should be important for the development of new nutritional strategies in malnourished patients with compromised intestinal functions.

Lysine nutrition is unique among indispensable amino acids in that it can be conserved and can be fed 12 h out of phase (delayed supplement) with the other dietary amino acids. In piglets, high levels (2-6%) of L-lysine added to a 10% protein diet can be tolerated without obvious detrimental effects. In both rat and piglet liver preparations, the first enzyme in the saccharopine-dependent pathway of lysine catabolism, lysine [alpha]-ketoglutarate reductase (LKR), is found only in the mitochondrial matrix. For Lys catabolism to occur, Lys must first enter the matrix of the mitochondrion. LKR, saccharopine dehydrogenase, mitochondrial lysine uptake, and lysine oxidation (LOX) all increased >3-fold in rats fed high levels of dietary protein (up to 60%). The activities of mitochondrial Lys uptake and LOX were similar when expressed as mmol/(d * 100 g body weight). Thus, LOX can be a proxy for mitochondrial Lys uptake. Piglet liver LKR and LOX increase 5- to 10-fold when piglets are fed high-protein (50 or 75%) diets. In both the rat and piglet, after adapting to the high protein diet, the activity of LKR is 400-500 times that of LOX, suggesting that Lys uptake by a transporter(s) is rate limiting. Quantitative 24-h dietary infusion studies in piglets revealed that >80% of the Lys infused (4% of the diet) could not be recovered in the urine or body or accounted for by calculated Lys oxidation based on liver activity of LOX. Other pathways and tissues may account for the Lys oxidation in piglets.

Arginine has multiple metabolic fates and thus is one of the most versatile amino acids. Not only is it metabolically interconvertible with the amino acids proline and glutamate, but it also serves as a precursor for synthesis of protein, nitric oxide, creatine, polyamines, agmatine, and urea. These processes do not all occur within each cell but are differentially expressed according to cell type, age and developmental stage, diet, and state of health or disease. Arginine metabolism also is modulated by activities of various transporters that move arginine and its metabolites across the plasma and mitochondrial membranes. Moreover, several key enzymes in arginine metabolism are expressed as multiple isozymes whose expression can change rapidly and dramatically in response to a variety of different stimuli in health and disease. As illustrated by the questions raised in this article, we currently have an imperfect and incomplete picture of arginine metabolism for any mammalian species. It has become clear that a more complete understanding of arginine metabolism will require integration of information obtained from multiple approaches, including genomics, proteomics, and metabolomics.

Diet-induced (taurine deficiency) dilated cardiomyopathy is reported more in large than small dogs possibly because taurine biosynthesis rate (TBR) is lower in large than small dogs. The TB R in 6 mongrels (37.9 [+ or -] 2.1 kg) and 6 beagles (12.8 [+ or -] 0.4 kg) was determined from the fractional dilution rate of urinary [[1,2-.sup.2] [H.sub.2]]-taurine, (d4-tau). All dogs were given a 15.6% protein, 0.60% sulfur amino acid (SAA) diet in amounts to maintain an ideal body condition score. After 3 mo, 14.6 mg/kg body weight of d4-tau was given orally and TBR determined from d4-tau to taurine ratio in urine collected each d for 6 d. Enrichments of d4-tau were determined by GC-MS. Thereafter, mongrels and beagles were paired by ranking of SAA intake per metabolic body weight per [kg.sup.0.75] Each pair received the same amount of diet/kg [kg.sup.0.75] for 2 wk, then TBR was again determined. Concentrations of taurine in plasma, blood, and urine and concentrations of plasma thiols were measured during each TBR determination. In Expt. 1, TBR and taurine concentrations in plasma and urine of mongrels were lower (P< 0.05) than those of beagles. In Expt. 2, TBR and taurine concentrations in blood and plasma of mongrels were lower (P< 0.05) than beagles. Together, the results support the hypothesis that large compared with small dogs have lower TBR when fed diets nearlimiting in dietary SAA, but adequate to maintain ideal body condition.

To determine the effects of chronic maternal renal insufficiency on fetal renal function, we studied nine fetuses whose mothers underwent subtotal nephrectomy at least 2 mo before mating (STNxF) and seven fetuses from intact ewes (IntF) (126-128 days of gestation, term 150 days). STNxF had lower hematocrit (P < 0.05), plasma chloride (P < 0.01), and creatinine levels (P < 0.01), and the length-to-width ratio of their kidneys was reduced (P < 0.05). They excreted twice as much urine (P < 0.05) and sodium (P < 0.01). Total (P = 0.01) and proximal fractional sodium reabsorptions (P < 0.05) were lower in STNxF; distal delivery of sodium (P < 0.05) and distal fractional sodium reabsorption (P < 0.05) were higher. They tended to have suppressed renin levels (P = 0.06). Infusions of amino acids (alanine, glycine, proline, and serine at 0.32 mmol/min for 1 h and 0.64 mmol/min for 2 h intravenously), known to stimulate renal blood flow and glomerular filtration rate in fetal sheep, did so in IntF (P < 0.01). Arterial pressure also increased (P < 0.01). These effects were not observed in STNxF. In summary, chronic maternal renal insufficiency was associated with profound alterations in fetal renal excretion of fluid and electrolytes and impaired renal hemodynamic and glomerular responses to amino acid infusion. Whether these marked changes in the renal function of fetuses carried by STNx ewes are associated with alterations in renal function in postnatal or adult life remains to be determined. glomerular filtration; amino acid infusions; hypochloremia; hyponatremia; renin