The short-term feeding of low- and high-histidine diets prior to market affects the muscle carnosine and anserine contents and meat quality of broilers.

Functional dipeptides carnosine and anserine are abundant in muscle. We determined the effect of short-term dietary histidine (His) content on muscle carnosine and anserine contents and meat quality of broilers. Three groups of 28-day-old female broilers were fed diets with His contents of 67%, 100%, or 150% of requirement for 10 days before market (His contents 0.21%, 0.32%, and 0.48%, respectively). The carnosine and anserine contents of 0-h aged muscle significantly increased with dietary His content; in particular, the carnosine content was 162% higher in the His 0.48% group than in the His 0.32% group. The contents of both peptides also increased with dietary His content in 48-h aged muscle, but carnosine was not detected in 0- and 48-h aged muscle of the His 0.21% group. The drip loss, cooking loss, shear force, and pH of meat were not affected by the dietary His content. The 2-thiobarbituric acid-reactive substances contents of 24- and 48-h aged muscles were lower in the His 0.48% group than in the other groups, and the a* and b* values were lower in the His 0.21% group. These results suggest that short-term dietary His content affects imidazole dipeptide contents, antioxidative capacity, and color of broiler meat.

Suppression of FoxO1 mRNA by ß2 -adrenoceptor-cAMP signaling through miR-374b-5p and miR-7a-1-3p in C2C12 myotubes.

ß2 -Adrenoceptor (ß2 -AR) signaling decreases the transcriptional activity of forkhead box O (FoxO), but the underlying mechanisms remain incompletely understood. Here, we investigated how ß2 -AR signaling regulates the protein abundance of FoxO and its transcriptional activity in skeletal muscle. We observed that stimulation of ß2 -AR with its selective agonist, clenbuterol, rapidly decreased FoxO1 mRNA expression, and this was accompanied by a decrease in either FoxO1 protein level or FoxO transcriptional activity. We subsequently observed that miR-374b-5p and miR-7a-1-3p were rapidly upregulated in response to ß2 -AR stimulation. Transfection with mimics of either miRNA successfully decreased FoxO1 mRNA. Moreover, because ß2 -AR stimulation increased cAMP concentration, pretreatment with an adenylyl cyclase inhibitor canceled out these effects of ß2 -AR stimulation. These results suggest that ß2 -AR stimulation results in rapid upregulation of miR-374b-5p and miR-7a-1-3p in myotubes, which in turn results in a decrease in FoxO1 mRNA expression via the ß2 -AR-cAMP signaling pathway.

Reduction in dietary lysine increases muscle free amino acids through changes in protein metabolism in chickens.

Taste is crucial to meat quality, and free Glu is an important taste-active component in meat. Our recent study showed that the short-term feeding of a low-Lys diet increases the concentration of free Glu and other free amino acids in chicken muscle and improves its taste. Here, we investigated the mechanisms by which the feeding of a low-Lys diet increases free Glu in chicken muscle. Two groups (n = 10 per group) of 28-day-old female Ross strain broiler chickens were fed diets with a graded Lys content of 90% or 100% of the recommended Lys requirement (according to National Research Council [1994] guidelines) for 10 D. Free amino acid concentrations and the mRNA abundance of protein metabolism-related genes were measured in breast muscle, and breast muscle metabolome analysis was conducted. Free Glu in muscle was increased by 51.8% in the Lys 90% group compared with the Lys 100% group (P < 0.01). Free threonine, glutamine, glycine, valine, isoleucine, leucine, tyrosine, phenylalanine, histidine, and 3-methyl-histidine concentrations in breast muscle were also increased in the Lys 90% group (P < 0.05). Metabolome analysis also showed that free amino acids were increased in the Lys 90% group. The mRNA abundance of µ-calpain, caspase-3, and 20S proteasome C2 subunit were increased in the Lys 90% group (P < 0.05). Moreover, the free Glu concentration in muscle was correlated with mRNA abundance of µ-calpain (r = 0.74, P < 0.01), caspase 3 (r = 0.69, P < 0.01), 20S proteasome C2 subunit (r = 0.65, P < 0.01), and cathepsin B (r = 0.52, P < 0.05). Our study suggests that the feeding of a low-Lys diet to chickens increased the free Glu content of breast muscle by promoting protein degradation.

Evaluation of Postprandial Glycemic Response and Physical Properties of High-Amylose Rice "Koshinokaori".

This study evaluated the postprandial glycemic response and physical properties of the high-amylose rice, Koshinokaori (KK), cooked under different conditions. Twelve healthy subjects (Japanese, 6 males, 6 females) were given cooked, white KK rice or tomato chicken rice (TCR) using KK rice. The Japanese standard rice, Koshihikari (KH), was used as reference. All meals contained the same amount (50 g) of available carbohydrate. Blood glucose levels were measured at 0 (fasting), 15, 30, 45, 60, 90, and 120 min after each meal. The results from the cooked, white KK rice showed a significant difference in blood glucose variation at 60, 90, and 120 min and the incremental area under the curve (IAUC) of blood glucose concentration for KK cooked at optimal water to rice ratio was observed. Blood glucose variation and IAUC after intake of TCR-KK rice was lower than that after TCR-KH rice intake. Addition of 5% trehalose to KK rice resulted in a smaller decrease in adhesiveness and stickiness of cooked rice after 180 min at 20ºC. The addition of 5% trehalose to KK rice also produced favorable results in the sensory evaluation. KK rice produces favourable postprandial glycemic responses and physical properties under varied cooking condition and thus, may be beneficial in the prevention of lifestyle-related diseases such as type 2 diabetes.

Dietary Supplementation with Lysine and Threonine Modulates the Performance and Plasma Metabolites of Broiler Chicken.

Here, we investigated whether the optimal threonine (Thr) to lysine (Lys) ratio in high Lys diet improves the growth performance of modern broiler chickens at finisher period and determined the possible mechanism underlying improvement in the growth performance of chickens fed with high Lys or Lys + Thr diet using metabolome analyses. Eighteen 21-day-old chickens housed in individual cages were randomly divided into three groups of six chickens fed with different diets as follows: control diet, high Lys diet (150% Lys content of National Research Council requirement), and high Lys + Thr diet (0.68 of Thr/Lys in high Lys diet). Body weight gain (BWG) increased in chickens receiving high Lys diet as compared with those fed with the control diet (P<0.05); no significant difference was observed in BWG of chickens from high Lys + Thr and high Lys groups. Feed conversion ratio (FCR) was lower in chickens fed with high Lys or high Lys + Thr diet than in those on the control diet. Serotonin concentration increased in the plasma of chickens fed with high Lys diet as compared to those fed with other diets. A negative correlation was observed between plasma serotonin concentration and FCR. These results provide the first evidence on the use of high Lys in broiler diets to reduce FCR during finisher period, which may be associated with change in plasma serotonin concentration. These findings suggest that high Lys content in finisher diet, but not high Thr + Lys diet, may affect the peripheral serotonergic metabolism and improve FCR. Thus, plasma serotonin may serve as a biomarker of FCR in broilers.

Descriptive Sensory Traits of Cooked Eggs Laid from Hens Fed Rice Grain.

Descriptive sensory characteristics of eggs produced by conventional corn-based feeding and unhulled whole rice grain-feeding were compared in two cooking procedures using a trained panel. Rice-feeding significantly decreased brothy and roasted odor in eggs cooked into half-cooked egg yolks, and decreased the creamy odor, smoothness and moisture of eggs cooked into custard puddings. However, a statistical interaction between rice-feeding and production farm was not observed in every sensory attribute. These findings indicated that replacing corn with unhulled whole rice grain in diets for laying hens alters the sensory attributes of eggs.

Influence of Varying Dietary Protein Levels on Glycation of Albumin, Tryptophan and Valine in the Plasma of Chickens.

Glycation is a chemical reaction in which reducing sugars bind non-enzymatically to compounds containing amino groups. Avian species like chickens are hyperglycemic animals and have high body temperature compared to mammalian species, which enables avian species to accelerate the glycation of proteins and amino acids with glucose. Although varying dietary crude protein (CP) levels alter plasma concentrations of proteins and amino acids, the influence of varying CP levels on the glycation of plasma proteins and amino acids has not been studied so far. In the present study, therefore, glycation of albumin, tryptophan and valine in the plasma of chickens fed diets with varying CP levels (0, 10, 20, 40 and 60%) was examined. At the end of the experimental period, blood samples were collected and plasma concentrations of glycoalbumin, glycated tryptophan (tryptophan-Amadori product and (1R, 3S) - 1 - (D - gluco - 1, 2, 3, 4, 5 - pentahydroxypentyl) - 1, 2, 3, 4 - tetrahydro - ß - carboline - 3 - carboxylic acid (PHP-THßC)), and valine-Amadori product were measured. Although plasma albumin concentration was reduced along with the decrease in dietary CP levels from 20% to 0%, glycoalbumin in the plasma was increased under such dietary conditions. Similar increase in the ratios of tryptophan-Amadori product to tryptophan and valine-Amadori product to valine in the plasma of chickens fed a protein-free diet was observed. These results suggest that dietary protein deficiency might enhance the non-enzymatic glycation of plasma proteins and amino acids in chickens.

Reduction of dietary lysine increases free glutamate content in chicken meat and improves its taste.

Taste is a crucial factor of meat quality, and amino acids are important taste-active components in meat. Here, the effects of dietary lysine (Lys) content on taste-active components in meat, especially free glutamate (Glu), were investigated. Twenty-eight-day-old broilers (Gallus gallus) were fed diets with graded Lys content of 90% or 100% of the recommended Lys requirement, (according to the National Research Council, ) for 10 days. Free amino acid content in meat and sensory scores of meat soup were estimated. Free Glu content, the main taste-active component of meat, was significantly increased by a reduction of dietary Lys. Compared with the Lys 100% group (control), free Glu concentrations of meat were increased by 35.7% in the Lys 90% group (P < 0.05). In addition, free glycine, valine, isoleucine, leucine, histidine and threonine concentrations of meat were significantly increased in the Lys 90% group (P < 0.05). Sensory evaluation of meat soup made from the Lys 100% and 90% groups indicated different meat tastes. Sensory scores of taste intensity, umami and kokumi tastes were significantly higher in the Lys 90% group. These results suggest that a reduction of dietary lysine increased free glutamate content in meat and improved its taste.

Rice endosperm protein slows progression of fatty liver and diabetic nephropathy in Zucker diabetic fatty rats.

We previously reported that rice endosperm protein (REP) has renoprotective effects in Goto-Kakizaki rats, a non-obese diabetic model. However, whether these effects occur in obese diabetes remains unclear. This study aimed to clarify the effects of REP on obese diabetes, especially on fatty liver and diabetic nephropathy, using the obese diabetic model Zucker diabetic fatty (ZDF) rats. In total, 7-week-old male ZDF rats were fed diets containing 20 % REP or casein (C) for 8 weeks. Changes in fasting blood glucose levels and urinary markers were monitored during the experimental period. Hepatic lipids and metabolites were measured and renal glomeruli were observed morphologically. HbA1c levels were significantly lower in rats fed REP, compared with C (P<0·05). Compared with C in the liver, REP prevented lipid accumulation (total lipid, TAG and total cholesterol, P<0·01). Liver metabolome analysis indicated that levels of metabolites associated with glycolysis, the pentose phosphate pathway and carnitine metabolism were significantly greater in the REP group than in the C group (P<0·05), suggesting activation of both glucose catabolism and fatty acid oxidation. The metabolite increases promoted by REP may contribute to suppression of liver lipid accumulation. Urinary excretion of albumin and N-acetyl-ß-d-glucosaminidase was significantly reduced in rats fed REP for 8 weeks (P<0·01). In addition, there was a distinct suppression of mesangial matrix expansion and glomerular hypertrophy in response to REP (P<0·01). Thus, REP had preventive effects on obese diabetes, fatty liver and diabetic nephropathy.

Protective effect of composite earthworm powder against diabetic complications via increased fibrinolytic function and improvement of lipid metabolism in ZDF rats.

Thrombosis is the leading cause of mortality globally. It is not only a complication but also a risk factor for progression of diabetes. However, alternative oral therapies and prophylaxis with less adverse effect for thrombosis have not been well studied. In this study, composite powder containing earthworm (CEP) was used and its fibrinolytic activity was measured. CEP was found to have a high urokinase-type plasminogen activator like activity in an in vitro assay. It also had significantly shortened euglobulin clot lysis time (ECLT) at 4 and 24 h after ingestion in Sprague Dawley rats. Zucker Diabetic Fatty rats were used to assess the effect of CEP on diabetes and diabetic nephropathy. After 10 weeks of feeding, CEP significantly shortened ECLT and attenuated HbA1c, hepatic lipid accumulation, and urinary albumin excretion and improved glomerular mesangial matrix score. Therefore, CEP may have beneficial effects on diabetes and diabetic nephropathy.

Effect of dietary histidine on contents of carnosine and anserine in muscles of broilers.

Carnosine (ß-alanyl-L-histidine) and anserine (ß-alanyl-1-methyl-L-histidine) are dipeptides mainly found in skeletal muscle and brain of many vertebrates, and particularly high concentrations are observed in chicken pectoral muscles. It was reported that these peptides have many functions, such as antioxidant activity. In this study, we examined the effect of different levels of dietary histidine on carnosine and anserine contents in broiler muscles. The 14-days-old female Chunky strain broilers were given feeds containing three different levels of histidine; 67% (Low-His), 100% (Control) and 200% (High-His) of histidine requirement according to the NRC (1994). Chicks were fed experimental diets for 10 days. Both dipeptides in muscle were significantly decreased. In particular, carnosine was not detected at all in the Low-His group and was significantly increased in the High-His group. Both dipeptides were not detected in plasma. These results indicated the possibility to produce chicken meat with enhanced amount of these dipeptides by high histidine feeding.

Regulation of free glutamate content in meat by dietary lysine in broilers.

Regulation of taste is important for improving meat quality and glutamate (Glu) is one of the important taste-active components in meat. Here, the effects of dietary lysine (Lys) content on taste-active components in meat, especially free Glu, were investigated. Fourteen-day-old broiler chicks (Gallus gallus) were fed on diets containing 100% or 150% of the recommended Lys content for 10 days. Concentrations of free amino acids in plasma, muscle and liver were measured. The levels of messenger RNAs (mRNAs) for enzymes related to Glu metabolism were determined in muscle and liver. The concentration of muscle metabolites was also determined. The free Glu content in muscle of chicks fed the Lys150% diet was increased by 44.0% compared with that in chicks fed the Lys100% diet (P < 0.01). The mRNA level of lysine α-ketoglutarate reductase, which is involved in Lys degradation and Glu production, was significantly increased (P < 0.05) in the Lys150% group. Metabolome analysis showed that the Lys degradation products, muscular saccharopine, pipecolic acid and α-aminoadipic acid, were increased in the Lys150% group. Our results suggest that free Glu content in muscle is regulated by Lys degradation. These results suggest that a short-term feeding of high-Lys diet could improve the taste of meat.

In vivo digestibility of rice prolamin/protein body-I particle is decreased by cooking.

Rice has storage proteins, e.g., glutelin, globulin and prolamin, in the seeds, which are used as nitrogen sources during germination. Rice prolamin has been reported to be an indigestible protein that decreases the nutritional value of rice. However, the causes for the indigestibility of prolamin are currently not clear. The objective of this study was to determine if prolamin is naturally indigestible or if cooking affects its digestibility. The gastrointestinal (GI) transit of rice 23 kDa glutelin (23G) and 13 kDa prolamin (13P) in Wistar/ST rats fed raw rice (RR) and cooked rice (CR) diets was assessed using Western blot analysis. We also measured the excretion of these proteins in the feces of these rats. Additionally, morphological observation of the structure of type-I protein bodies in the feces was performed using electron microscopy. Assessment of GI transit revealed that 23G rapidly disappeared from the GI contents of both the RR and CR groups, but 13P accumulated in the cecum of the CR group. In the CR group, prolamin, maintaining the structure of PB-I, was fully excreted in the feces. These results indicate that rice prolamin is not indigestible by nature, but is rendered indigestible by cooking.

Effect of dietary astaxanthin rich yeast, Phaffia rhodozyma, on meat quality of broiler chickens.

We evaluated effects of dietary supplementation with astaxanthin (Ax)-rich yeast, Phaffia rhodozyma (Xanthophyllomyces dendrorhous), on broiler chicken meat quality. Fourteen-day-old female Ross broilers were divided into three groups: control group, Ax-free diet; Ax 10 group, 10 mg/kg Ax diet; and Ax 20 group, 20 mg/kg Ax diet for 28 days. At 42 days old, chickens were slaughtered, and then growth performance, meat quality and sensory attributes were analyzed. Compared with the control, a* values increased significantly after slaughter and 48 h postmortem for Ax 20 samples (P<0.05) and for b* values in Ax 20 and Ax 10 groups (P<0.05). Cooking loss decreased in the Ax 20 group (P<0.05). After 120 h aging, contents of several free amino acids and total free amino acid content of Ax 20 group were significantly higher than the control (P<0.05). In sensory evaluation, meat texture attributes improved significantly in the Ax 20 group (P<0.01). No significant changes occurred in flavor attribute scores of meat soup from the Ax 20 group compared with the control even though most assessors preferred meat soup from the Ax 20 group. Overall, Ax-rich yeast in the diet improves broiler chicken meat quality.

Diversity of amino acid signaling pathways on autophagy regulation: a novel pathway for arginine.

Autophagy is the intracellular bulk degradation process to eliminate damaged cellular machinery and to recycle building blocks, and is crucial for cell survival and cell death. Amino acids modulate autophagy in response to nutrient starvation and oxidative stress. We investigated the relevance of reactive oxygen species (ROS) production on the regulation of autophagy using amino acids, both as a mixture and individually, in rat hepatoma H4-II-E cells. Nutrient starvation elevated ROS production and stimulated autophagy. Treatment with complete (CAA), regulatory (RegAA) and non-regulatory (NonRegAA) amino acid mixtures showed significant suppression of ROS production, whereas only CAA and RegAA exhibited significant suppression of autophagy, suggesting a dissociation of the two responses. The effects of individual amino acids were examined. Leucine from RegAA decreased ROS production and suppressed autophagy. However, methionine and proline from RegAA and arginine, cystine and glutamic acid from NonRegAA suppressed autophagy with an opposite increase in ROS production. Other amino acids from the NonRegAA group showed stimulating effects on ROS production without an autophagic response. Arginine's effect on autophagy suppression was not blocked by rapamycin, indicating an mTOR-independent pathway. Inhibitor studies on arginine-regulated autophagy may indicate the involvement of NO pathway, which is independent from ROS and mTOR pathways.

Effect of dietary vitamin E on broiler meat qualities, color, water-holding capacity and shear force value, under heat stress conditions.

This study was conducted to evaluate the effect of dietary vitamin E (VE) on broiler meat quality, especially focused on PSE (pale color, soft and exudative), under chronic heat stress (HS) conditions. Twenty-eight-day-old female Ross broilers were kept in independent cages with a controlled temperature of 24°C (normal temperature: NT) or 30°C (high temperature: HT). The NT chickens were fed basal feed. The HT chickens were fed basal feed (HT) or VE (200 mg/kg) added feed (HT + E). Broilers were weighed and slaughtered at 38 days old. The breast muscle was removed immediately and then the samples were used for determination of meat color, pH, water holding capacity (WHC) and shear force value (SFV). Body weight gain and feed intake were significantly decreased in the HT and HT + E groups compared to the NT group. VE supplementation did not affect the growth performance. Chronic HS at 30°C for 10 days may cause deterioration of meat quality such as PSE. The effects of chronic HS on meat quality were most significant in the toughness of broiler breast meat. Supplementation of VE in broiler feed would be effective to prevent the extent of PSE on broiler meat by chronic HS.

Rice protein ameliorates the progression of diabetic nephropathy in Goto-Kakizaki rats with high-sucrose feeding.

The effect of rice protein (RP) on diabetic nephropathy in non-obese, spontaneous type 2 diabetic Goto-Kakizaki (GK) rats was investigated.GK rats at 7 weeks of age were fed 20% RP or casein (C) in standard or high-sucrose diets for 10 weeks. Plasma total cholesterol,TAG, alkaline phosphatase (ALP), adiponectin, creatinine and urinary albumin excretion (UAE) were measured and renal histology was evaluated. Compared with C, RP lowered plasma TAG and improved plasma adiponectin levels in GK rats fed the standard diet (P<0·05), and also lowered total cholesterol and ALP in high-sucrose-fed GK rats (P<0·05). RP markedly suppressed the sharp increase in UAE when GK rats were fed high-sucrose diets (P<0·05), and prevented glomerular mesangial matrix expansion in the deep renal cortex near the corticomedullary junction (P<0·05). These results strongly indicate that dietary RP can ameliorate the progression of diabetic nephropathy at an early stage compared with C.

Regulation of muscular glutamate metabolism by high-protein diet in broiler chicks.

The major taste active component, glutamate (Glu), improves the taste of meat. In this study, we investigated the effect of a short-term high-protein (HCP) diet on the intramuscular free Glu content to improve the taste of meat. Furthermore, we elucidated how the muscle free Glu content was controlled by the HCP diet. Chicks (14 days old) were fed the control diet or HCP diet for 10 days. Plasma and muscle free amino acid concentrations, and activity and messenger RNA (mRNA) expression of muscle enzymes related to Glu metabolism were determined. Muscle free Glu content was increased (P < 0.01) by 51%. Activity and mRNA expression of glutaminase (GA), which is one of the major Glu-related enzymes, were significantly decreased (P < 0.05) in the HCP group because of feedback inhibition. The mRNA expression of lysine α-ketoglutarate reductase (LKR), which is the enzyme involved in lysine (Lys) degradation and Glu production, was significantly increased (P < 0.001) in the HCP group. These results suggest that short-term dietary HCP feeding is an effective treatment for improving the taste of meat. Furthermore, our results suggest that the free Glu content in muscle is regulated by GA and LKR.

Beef texture characterization using internationally established texture vocabularies in ISO5492:1992: differences among four different end-point temperatures in three muscles of Holstein steers.

'Tenderness' has been an important sensory characteristic for beef, although 'tenderness' has not been commonly defined. On the other hand, ISO5492:1992 provides internationally established vocabularies for sensory analysis with simple definition. The aim of this study was texture characterization for three beef muscles cooked to four end-point temperatures using ISO5492:1992 texture terms in Japanese to develop objective sensory evaluation terms for beef texture other than 'tenderness.' Longissimus, semitendinosus, and psoas major muscles harvested from three Holstein steers were cooked to 45, 60, 72, and 92 degrees C end-point temperatures and evaluated by a trained sensory panel. Correspondence analysis indicated that the 'chewiness' and 'hardness' defined in ISO5492 were distinguished in each muscle. Changes in the 'chewiness' and 'hardness' qualities during cooking were different from each other. These findings suggest that both 'chewiness' and 'hardness' as defined in ISO5492:1992 should be evaluated simultaneously to determine the sensory texture of beef. Warner-Bratzler shear force values (WBSFVs) were also correlated with ISO5492 'chewiness.' This finding suggests that WBSFV indicates ISO5492 'chewiness' rather than undefined 'tenderness.'

Vitamin E as a novel enhancer of macroautophagy in rat hepatocytes and H4-II-E cells.

Autophagy is an intracellular bulk degradation process induced by nutrient starvation, and contributes to macromolecular turnover and rejuvenation of cellular organelles. We demonstrated that vitamin E was a novel nutritional enhancer of autophagy in freshly isolated rat hepatocytes and rat hepatoma H4-II-E cells. Supplementation of fresh hepatocytes with vitamin E (up to 100 microM) increased proteolysis significantly in the presence or absence of amino acids in a dose-dependent manner. The cytosolic LC3 ratio, a newly established index of autophagic flux, was significantly increased by vitamin E, strongly suggesting that the possible site of action is the LC3 conversion step, an early step in autophagosome formation. A typical antioxidant, alpha-lipoic acid, exerted autophagy suppression, while H(2)O(2) stimulated autophagy. It is conceivable that autophagy was stimulated by oxidative stress and this stimulation was cancelled by cellular antioxidative effects. However, in our studies, vitamin E could have enhanced autophagy over-stimulation by H(2)O(2), rather than suppress it. From these results, using a new cytosolic LC3 ratio, vitamin E increases autophagy by accelerating LC3 conversion through a new signaling pathway, emerging as a novel enhancer of autophagy.