Rat liver amino acid balances after the administration of an oral protein load.

The hepatic balances of amino acids, ammonia and urea have been measured in rats for three hours after receiving a protein load. The liver took up practically all of the portal ammonium. Alanine was retained to a large extent during all three hours. Other portal amino acids, mainly essential amino acids, were largely retained in about one hour after the gavage, to be released in a similar proportion thereafter. The other amino acids were also retained and then released, but to a lower extent. These amino acids were used in part by the liver for the synthesis and release of urea, which appearance in hepatic vein peaked at two hours after the protein administration.     

Gene and protein expression profiles in the foetal liver of the pregnant rat fed a low protein diet

Foetal growth is particularly sensitive to the protein content of the mother’s diet. Microarray data from the foetal liver of pregnant rats fed normal (HP) or reduced protein diets (LP) were compared by gene set enrichment analysis. Soluble proteins from a second portion of the liver were analysed by two-dimensional gel electrophoresis. Genes associated with progesterone, insulin-like growth factor-1 and vascular endothelial growth factor were upregulated in HP compared to LP, in addition to genes associated with cell differentiation and signalling from the extracellular matrix. In contrast, cytokine signalling was downregulated. Proteomics showed that proteins associated with amino acid metabolism, mitochondrial function and cell motility were differentially abundant in the HP compared to the LP groups. These growth factor and extracellular matrix signalling pathways linked to cell motility may be important mediators of the changes in liver structure that occur in utero and persist into adult life.     

Changes in UCP expression in tissues of Zucker rats fed diets with different protein content

The effect of dietary protein content on the uncoupling proteins (UCP) 1, 2 and 3 expression in a number of tissues of Zucker lean and obese rats was studied. Thirty-day-old male Zucker lean (Fa/?) and obese (fa/fa) rats were fed on hyperproteic (HP, 30% protein), standard (RD, 17% protein) or hypoproteic (LP, 9% protein) diets ad libitum for 30 days. Although dietary protein intake affected the weights of individual muscles in lean and obese animals, these weights were similar. In contrast, huge differences were observed in brown adipose tissue (BAT) and liver weights. Lean rats fed on the LP diet generally increased UCP expression, whereas the HP group had lower values. Obese animals, HP and LP groups showed higher UCP expression in muscles, with slight differences in BAT and lower values for UCP3 in subcutaneous adipose tissue. The mean values of UCP expression in BAT of obese rats were lower than in their lean counterpart, whereas the expression in skeletal muscle was increased. Thus, expression of UCPs can be modified by dietary protein content, in lean and obese rats. A possible thermogenic function of UCP3 in muscle and WAT in obese rats must be taken into account.     

Rat intestinal amino acid balances after the administration of an oral protein load.

The intestinal amino acid balances in rats given an oral load of protein were measured three hours after the gavage. During that period, about one half of the nitrogen from the protein given appeared as net balance in the portal blood. The release of amino acids was not a continuous process, since two peaks of maximal intestinal efflux were found at 1 and 2.5 hours after gavage. This pattern followed that of portal blood flow with a 30 minute delay. Under prandial conditions, the intestine showed a net uptake of glutamine, aspartate, serine, threonine, phenylalanine, tyrosine, leucine, isoleucine and valine. It also showed a net production of alanine, glutamate, ornithine skeleton (arginine + citrulline + ornithine) and ammonia. There was also a surge of taurine, attributed to reabsorption of secreted taurine conjugates. There was a net unchanged absorption of glycine, proline, lysine and histidine, with respect to their proportions in the protein administered. The results suggest that the amino acid metabolism in the intestine under prandial conditions is much less passive than is generally assumed. The intestinal action upon the luminal amino acids is not limited to absorption, but is directly implied in their transformation to complement the ensuing homeostatic action of the liver upon them.     

The effect of protein intake on the potential activity of the lysosomal vacuolar system in the cat.

1. Experiments were conducted to examine the relationship between protein intake and protein degradation in the liver of cats. 2. The cats were fed either a low protein/high carbohydrate diet (LP) or a high protein diet devoid of carbohydrate (HP). 3. The potential proteolytic activity of the lysosomal vacuolar system in the liver was assessed by both indirect (osmotic fragility of hepatic lysosomes) and direct (stereological measurement of lysosomal volume) methods. 4. The results from both tests indicated a significantly lower autophagic activity of the lysosomal system in the LP fed animals than in the HP fed cats. 5. This suppression of lysosomal protein degradation may represent an important mechanism for the conservation of proteins by the cat when low protein diets are fed.     

Effect of undegradable protein concentration in the post-weaning diet on body growth and reproductive development of Assaf rams

Twenty-four growing Assaf lambs, divided into four groups of six animals, were used to study the effect of the undegradable protein content of the post-weaning diet on feed intake, body growth and reproductive development. From week 1 to week 21, the four groups were fed ad libitum as follows: group LL was given barley straw and low protein concentrate (LP); group HH was given barley straw and high protein concentrate (HP); group LH was given barley straw and LP concentrate from week 1 to 11 (period 1) and barley straw and HP concentrate from week 12 to 21 (period 2); group HL was given barley straw and HP concentrate in period 1 and barley straw and LP concentrate in period 2. From week 22 to week 26 (period 3), all animals received the same amount of hay and LP concentrate. Barley straw intake was not significantly (P>0.05) affected by dietary treatments. In the 1st period, average concentrate intake and live body weight gain (LWG) were greater in lambs fed HP than LP supplement. In the 2nd period, concentrate intake was not significantly (P>0.05) affected by type of supplement, but LWG was greater for lambs fed HP than LP supplement. Scrotal circumference in week 11 was significantly (P<0.05) lower in lambs fed LP supplement than in lambs fed HP supplement. No significant differences (P>0.05) due to dietary treatments were observed on scrotal circumference in weeks 21 and 25. Dietary treatments had no significant (P>0.05) effect on either circulating concentration of testosterone or ejaculate characteristics. In conclusion, results from this study suggest that supplementing diets with undegradable protein enhanced performance throughout the breeding period and accelerated testis growth. Nevertheless, final testis size, pattern of circulating testosterone and sperm output were unaffected by dietary treatments.     

饵料蛋白水平对德国小蠊肠道细菌群落的影响

昆虫肠道微生物在宿主营养代谢、生长发育、免疫以及抵御病原菌等方面具有重要作用。研究不同蛋白水平饵料饲养对德国小蠊（Blattella germanica）雄成虫肠道细菌群落组成及其功能的作用，探究德国小蠊肠道细菌对宿主营养和健康的影响，以期为发展生物防治的诱食性饵料提供理论支持。分别取连续饲喂低蛋白（LP2组：5%）、高蛋白（HP3组：65%）以及正常蛋白水平饵料（CD1组：25%）21 d的德国小蠊雄成虫，饥饿24 h后无菌条件下分离并提取肠道总基因组DNA，采用特异引物扩增细菌16S rDNA的V4可变区并进行高通量测序，分析德国小蠊肠道细菌群落组成及其功能特征。结果表明德国小蠊肠道细菌主要由拟杆菌门（Bacteroidetes）、变形菌门（Proteobacteria）、梭杆菌门（Fusobacteria）和厚壁菌门（Firmicutes）等细菌群落组成。饲喂低蛋白饵料LP2组的德国小蠊肠道细菌中拟杆菌属（Bacteroides）细菌丰度（47.44%）显著高于HP3组（23.97%）和对照CD1（7.04%）。饲喂高蛋白饵料的HP3组梭杆菌门丰度显著高于其他两组。LEfSe物种差异分析也表明HP3组德国小蠊肠道细菌中梭杆菌属（Fusobacterium）细菌与低蛋白饵料LP2组和对照CD1组有显著差异。基于Tax4Fun功能预测显示，HP3高蛋白饵料组的德国小蠊肠道细菌中与能量代谢功能相关基因的相对丰度极显著高于对照组CD1组，外源性物质代谢与降解和其他氨基酸代谢功能基因的相对丰度显著高于低蛋白LP2组。本研究结果表明饵料中蛋白质水平的差异能够显著改变德国小蠊肠道中细菌群落结构组成，并影响其代谢功能。     

The level of dietary protein and carbohydrate has a different effect on intestinal uptake of hexoses and lipids in rabbits with an ileal resection than in those with an intact intestinal tract

Either high protein, low carbohydrate (HP) or low protein, high carbohydrate (LP) diets were fed for 6 weeks to rabbits with or without resection of the distal half of the small intestine. Control and resected rabbits fed HP consumed more food and gained more weight than the animals fed LP. The level of dietary protein has a different effect on intestinal transport in animals with an ileal resection than in those with an intact small intestine. With 0.5 mM glucose, the in vitro uptake in control rabbits was greater for the LP than HP diet but was unchanged in resected rabbits; uptake of 0.5 mM galactose and 3-O-methyl glucose was unaffected by HP and LP, whereas in rabbit uptake was lower in LP than HP. The uptake of 40 mM glucose was greater with the LP than HP diet in control rabbits, but lower with LP than HP in resected rabbits. In control rabbits, the uptake of aluric acid was lower on the LP than HP diet; the uptake of octanoic acid, decanoic acid, and cholesterol was unchanged; and the uptake of each fatty acid and cholesterol was greater in resected rabbits fed LP than HP. Feeding a low protein diet to animals with an ileal resection is associated with lower jejunal uptake of high concentrations of glucose, but the higher uptake of galactose and enhanced permeability to fatty acids result in superior weight gain. Thus, recommendations for alterations in dietary protein and carbohydrate levels following ileal resection must be made with the knowledge that these changes may influence intestinal transport function as well as body weight gain.     

Effect of protein intake on amino acid catabolism and gluconeogenesis by isolated hepatocytes from the cat (Felis domestica)

Cats were fed 17.5% (LP) and 70% (HP) diets and hepatocytes were prepared from them. Rates of gluconeogenesis from pyruvate, alanine and threonine (10 mM) were unaffected by protein intake but 10 mM glutamine was converted faster by cells from HP fed animals. Rates of oxidation of alanine, threonine and glutamine and flux rates of tyrosine aminotransferase and tryptophan 2,3-dioxygenase were greater in cells from HP fed cats at all amino acid concentrations used. Proteolysis was indicated by urea production which was higher in cells from HP fed cats but was reduced significantly by leupeptin.     

Effects of feeding low protein diets to piglets on plasma urea nitrogen, faecal ammonia nitrogen, the incidence of diarrhoea and performance after weaning

This study evaluated the effects of feeding pigs low protein (LP) diets for different lengths of time after weaning on indices of protein fermentation, the incidence of postweaning diarrhoea (PWD), growth performance, and total-tract apparent digestibility. Sixty weaner pigs weighing 6.1 +/- 0.13 kg (mean +/- SEM) were used in a completely randomised design having five treatments: (i) a high protein diet (HP, 243 g/kg CP) fed for 14 d after weaning (HP14); (ii) a low protein diet (LP, 173 g CP/kg) fed for 5 d after weaning (LP5); (iii) LP diet fed for 7 d after weaning (LP7); (iv) LP diet fed for 10 d after weaning (LP10), and (v) LP diet fed for 14 d after weaning (LP14). All diets were supplemented with lysine, methionine, tryptophan and threonine, with all LP diets additionally fortified with crystalline isoleucine and valine to conform to a proposed ideal amino acid (AA) pattern. A second-stage diet (215 g CP/kg) was fed to pigs at the conclusion of each treatment. None of the diets contained antimicrobial compounds. Feeding a LP diet, regardless of duration of feeding, decreased plasma urea nitrogen (p < 0.001) and faecal ammonia-nitrogen (p < 0.001) contents. Feeding a LP diet, irrespective of feeding duration, decreased the incidence of PWD at day 8 after weaning (p = 0.044), and pigs fed diets LP7, LP010 and LP14 had firmer faeces (p = 0.030, p = 0.047 and p = 0.007, respectively) between days 10 and 12 after weaning. Treatments LP5, LP7, LP10 and LP14 did not reduce (p > 0.05) growth performance up to 106 days after weaning compared to pigs fed the HP diet. Total-tract apparent digestibility of dry matter, energy and crude protein were similar (p > 0.05) between treatments. Our data suggest that feeding a LP diet, supplemented with AA to conform to an ideal AA pattern, for 7-10 days after weaning can reduce PWD in pigs fed antibiotic-free diets without compromising production.     

Intrauterine growth retarded progeny of pregnant sows fed high protein:low carbohydrate diet is related to metabolic energy deficit

High and low protein diets fed to pregnant adolescent sows led to intrauterine growth retardation (IUGR). To explore underlying mechanisms, sow plasma metabolite and hormone concentrations were analyzed during different pregnancy stages and correlated with litter weight (LW) at birth, sow body weight and back fat thickness. Sows were fed diets with low (6.5%, LP), adequate (12.1%, AP), and high (30%, HP) protein levels, made isoenergetic by adjusted carbohydrate content. At -5, 24, 66, and 108 days post coitum (dpc) fasted blood was collected. At 92 dpc, diurnal metabolic profiles were determined. Fasted serum urea and plasma glucagon were higher due to the HP diet. High density lipoprotein cholesterol (HDLC), %HDLC and cortisol were reduced in HP compared with AP sows. Lowest concentrations were observed for serum urea and protein, plasma insulin-like growth factor-I, low density lipoprotein cholesterol, and progesterone in LP compared with AP and HP sows. Fasted plasma glucose, insulin and leptin concentrations were unchanged. Diurnal metabolic profiles showed lower glucose in HP sows whereas non-esterified fatty acids (NEFA) concentrations were higher in HP compared with AP and LP sows. In HP and LP sows, urea concentrations were 300% and 60% of AP sows, respectively. Plasma total cholesterol was higher in LP than in AP and HP sows. In AP sows, LW correlated positively with insulin and insulin/glucose and negatively with glucagon/insulin at 66 dpc, whereas in HP sows LW associated positively with NEFA. In conclusion, IUGR in sows fed high protein:low carbohydrate diet was probably due to glucose and energy deficit whereas in sows with low protein:high carbohydrate diet it was possibly a response to a deficit of indispensable amino acids which impaired lipoprotein metabolism and favored maternal lipid disposal.     

Interactive Effects of Indigestible Carbohydrates,Protein Type,and Protein Level on Biomarkers of Large Intestine Health in Rats

The effects of indigestible carbohydrates, protein type, and protein level on large intestine health were examined in rats. For 21 days, 12 groups of six 12-week-old male Wistar rats were fed diets with casein (CAS), or potato protein concentrate (PPC), providing 14% (lower protein level; LP), or 20% (higher protein level; HP) protein, and containing cellulose, resistant potato starch, or pectin. Fermentation end-products, pH, and β-glucuronidase levels in cecal digesta, and ammonia levels in colonic digesta were determined. Cecal digesta, tissue weights, cecal and colon morphology, and colonocyte DNA damage were also analyzed. Digesta pH was lower, whereas relative mass of cecal tissue and digesta were higher in rats fed pectin diets than in those fed cellulose. Cecal parameters were greater in rats fed PPC and HP diets than in those fed CAS and LP diets, respectively. Short-chain fatty acid (SCFA) concentrations were unaffected by protein or carbohydrate type. Total SCFA, acetic acid, and propionic acid concentrations were greater in rats fed LP diets than in those fed HP. Cecal pool of isobutyric and isovaleric acids was greater in rats fed PPC than in those fed CAS diets. PPC diets decreased phenol concentration and increased ammonia concentration in cecal and colonic digesta, respectively. Cecal crypt depth was greater in rats fed PPC and HP diets, and was unaffected by carbohydrates; whereas colonic crypt depth was greater in rats fed cellulose. Myenteron thickness in the cecum was unaffected by nutrition, but was greater in the colon of rats fed cellulose. Colonocyte DNA damage was greater in rats fed LP diets than in those fed HP diets, and was unaffected by carbohydrate or protein type. It was found that nutritional factors decreasing cecal digesta weight contribute to greater phenol production, increased DNA damage, and reduced ammonia concentration in the colon.     

Increased dietary protein elevates plasma uric acid and is associated with decreased oxidative stress in rapidly-growing broilers

Uric acid is an important antioxidant and methods to elevate its plasma concentration may be important in animal health. In a first study, the effect of dietary protein on plasma uric acid (PUA) and glucose concentrations were determined in 3-week-old chicks. Twenty-four broiler chicks were randomly assigned to four diets: a commercial control diet (C, 20% crude protein), low protein (LP) containing 10% casein, medium protein (MP) containing 20% casein or high protein (HP) containing 45% casein for a 3-week experiment. PUA concentration increased (P<0.05) in chicks fed HP diet and declined (P<0.05) in chicks fed LP while plasma glucose concentrations were lower (P<0.05) in chicks fed the LP diet at the end of the study. In a second study, PUA and leukocyte oxidative activity (LOA) were determined in broilers fed C, LP, MP or HP diets for 4 weeks. As in the first study, dietary protein directly affected PUA concentrations. In birds consuming HP diets, PUA was negatively correlated (P=0.06) with lowered LOA. These data support the view that increases in dietary protein can increase PUA concentrations, which can ameliorate oxidative stress.     

Exogenous and endogenous contributions to nitrogen fluxes in the digestive tract of pigs fed a casein diet. II. Ileal and faecal digestibilities and absorption of amino acids.

The present work aimed at quantifying nitrogen (N) and amino acid (AA) fluxes in the digestive tract of growing pigs fed a casein diet. In this paper we report on digesta passage at the terminal ileum, on apparent balances at the ileal and faecal levels, and on nutrients appearance in the portal vein. Digesta flow-rate at the terminal ileum was maximum between 6 and 12 h after the meal. About 10% of N and 5% of total AA ingested were recovered within 24 h. AA absorption started 30 min after the meal, and was measurable until 13 to 14 h. The total AA absorbed in 24 h accounted for 128% of the AA ingested. The AA composition of ileal digesta was very different from that of casein, closely resembling that of endogenous proteins. The AA composition of faeces was very close to that of bacterial proteins. The ileal digestibilities of AA, though lower than their faecal values, were very high. This was confirmed by AA absorption balances greater than 100%. These data suggest that casein was almost totally digested by the terminal ileum, and that endogenous AA were substantially reabsorbed. These findings are supported by data on endogenous N recycling (15N), reported in a following paper.     

Daily delivery of dietary nitrogen to the periphery is stable in rats adapted to increased protein intake

Dietary nitrogen was traced in rats adapted to a 50% protein diet and given a meal containing 1.50 g (15)N-labeled protein (HP-50 group). This group was compared with rats usually consuming a 14% protein diet and fed a meal containing either 0.42 g (AP-14 group) or 1.50 g (AP-50 group) of (15)N-labeled protein. In the HP group, the muscle nonprotein nitrogen pool was doubled when compared with the AP group. The main adaptation was the enhancement of dietary nitrogen transferred to urea (2.2 +/- 0.5 vs. 1.3 +/- 0.1 mmol N/100 g body wt in the HP-50 and AP-50 groups, respectively). All amino acids reaching the periphery except arginine and the branched-chain amino acids were depressed. Consequently, dietary nitrogen incorporation into muscle protein was paradoxically reduced in the HP-50 group, whereas more dietary nitrogen was accumulated in the free nitrogen pool. These results underline the important role played by splanchnic catabolism in adaptation to a high-protein diet, in contrast to muscle tissue. Digestive kinetics and splanchnic anabolism participate to a lesser extent in the regulation processes.     

Characterization and hormonal regulation of protein synthesis by the murine epididymis

Protein synthesis and secretion were examined in vitro by incubating minced tissue with [35S]methionine. The incorporation of label into tissue plus medium was linear for the 5 h of incubation. The percentage of available label incorporated into protein increased with the weight of tissue used. Approximately 13% of the label incorporated appeared in the medium after 5 h of incubation. Release of radioactive protein into the medium was characterized by an initial slow release (1-2 h) followed by a more rapid linear release between 3 and 5 h. Polyacrylamide gel electrophoresis revealed that the pattern of radioactive proteins present in the medium was different from and less complex than the tissue proteins. Substantial differences in protein patterns from different epididymal regions could be detected. The caput epididymidis was particularly active in secreting proteins characteristic of this region, whereas the corpus and cauda synthesized and secreted similar proteins. At least one of these proteins characteristic of the caput is stabilized by disulphide bonds. Short-term (9 day) castration resulted in reduced synthesis and secretion of several of these epididymal proteins. Testosterone administered after 9 days of castration reinitiated synthesis of some but not all of these epididymal proteins.     

Effects of dietary phosphorus intake on bone mineralization and calcium absorption in adult female rats

We investigated the effects of dietary phosphorus (P) intake on the bone mineralization and calcium (Ca) absorption in adult female rats. Fifteen 16-wk-old female Wistar rats were divided into three groups, and respectively fed a low-P diet containing 0.15% P (LP), a control diet containing 0.5% P (C), and a high-P diet containing 1.5% P (HP) for 42 d. The apparent Ca absorption was significantly increased with decreasing dietary P level. The serum parathyroid hormone concentration was significantly lower in the LP group than in the C and HP groups. The serum osteocalcin concentration and urinary excretion of deoxypyridinoline were significantly higher in the HP groups than in the LP and C groups. The bone mineral density of the fifth lumbar vertebra was significantly increased with decreasing dietary P level. These results indicate that the low-P diet increased Ca absorption, this being effective for bone mineralization in adult female rats.     

Radioglucose metabolism by richardson's ground squirrels in the weight-gain and weight-loss phases of the circannual cycle

Summary Captive fed, starved, and refed Richardson's ground squirrels in the weight-gain and weight-loss phases of the circannual cycle were injected with radioglucose and the activity of the label in skeletal muscle proteins and white adipose tissue lipids four hours after injection was used to determine if lean body mass and white adipose tissue would be rapidly restored when starved animals were refed. Starvation for six days reduced carcass mass 27–31% and white adipose tissue mass 23–24% (Table 1). Activity of the label in both tissues of weight-gain and weight-loss animals was reduced by starvation. After four days of refeeding activities retured to levels similar to those in fed animals, with the exception of lower activity in skeletal muscle proteins of weight-gain animals. Furthermore, activity in each tissue fraction of starved and refed weight-gain animals was similar to that in weight-loss animals when expressed as per cent of activity in the respective fed state (Table 2). Radioglucose incorporation indicated that when skeletal muscle and adipose tissue are depleted by starvation, distribution of the label upon refeeding is similar to that in the fed state. Four days after refeeding weight-gain phase ground squirrels had restored 5.5 g of lean body mass and 7.5 g of adipose tissue, including 1.4 g (6 kcal) of protein and 7.0 g (66 kcal) of lipid, respectively. These results are also consistent with the fed state, in which weight-gain animals were depositing more lipid than lean body mass.     

Na(+)-dependent transport of alanine and serine by liver plasma-membrane vesicles from rats fed a low-protein or a high-protein diet

Plasma-membrane vesicles prepared from the liver of rats fed either a low-(LP) or a high-protein (HP) diet exhibited Na(+)-dependent active transport of alanine and serine. The process gave apparent kinetic parameters compatible with a single saturable component for both amino acids. Na,K-ATPase (EC 3.6.1.37), marker of the basolateral domain of the hepatocyte plasma-membrane, was chosen as reference for the expression of amino acid transport in vesicle preparations. The high-protein diet induced a significant increase in liver Na,K-ATPase activity also found in corresponding plasma-membrane preparations, in parallel with an increase in the capacity towards amino acid transport. This suggests that in rats fed the high protein diet, transcellular Na+ exchange, although increased, remains well balanced. N-Methylaminoisobutyric acid (MeAIB), due to its poor velocity, proved unsuitable to distinguish between systems A and ASC in the experimental model. Comparing Na(+)- and Li(+)-driven transport, a family of carriers with strict Na(+)-dependency (A-like) was evidenced in LP vesicles but not in HP vesicles. The sensitivity to the lowering of the pH from 7.5 to 6.5 in the external medium was similar in both type of vesicles when Na+ was the driving ion. In the HP vesicles the Li(+)-tolerant, pH-insensitive component (ASC-like) was increased in parallel with overall Na(+)-dependent transport. These functional properties suggest that the carriers involved in the stimulation of transport in HP vesicles are composite in nature. Increasing concentrations of an amino acid mixture mimicking the changes of portal aminoacidemia inhibited the transport of alanine and of serine. The degree of inhibition was correlated with the relative concentration of substrate and was independent of the nutritional treatment.