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Polyamines (putrescine, spermidine, and spermine) are synthesized primarily from ornithine via ornithine decarboxylase (ODC) in mammals. Although avian tissues contain ODC activity, little is known about intracellular sources of ornithine for their polyamine synthesis. This study tested the hypothesis that arginase and proline oxidase contribute to polyamine synthesis in chickens. Kidney, jejunum, leg muscle, and liver from 0-, 7-, 14- and 21-day-old broiler chickens were assayed for the activities of arginase, proline oxidase (POX), ornithine aminotransferase (OAT), and ornithine decarboxylase (ODC). Kidney slices were also used to determine 14C-polyamine synthesis from [U-14C]arginine and [U-14C]proline. Furthermore, these tissues and plasma were analyzed for polyamines. Results indicate that all tissues contained OAT (mitochondrial) and ODC (cytosolic) activities, but arginase and POX activities were only detected in the mitochondria of chicken kidneys. Renal POX and arginase activities were greater at 7 days of age compared to newly hatched birds, and declined by Day 14. Renal arginase activity was greater at 21 days compared to 14 days of age, but there was no change in renal POX activity during that same period. Concentrations of polyamines in the kidneys and plasma were greater on Day 7 compared to Day 0 and decreased thereafter on Days 14 and 21. Kidney slices readily converted arginine and proline into polyamines, with peak rates being on Day 7. Concentrations of putrescine, spermidine and spermine in the plasma of chickens were about 20- to 100-fold greater than those in mammals. Our results indicate that polyamines are synthesized from arginine and proline in avian kidneys. Unlike mammals, polyamines released from the kidneys are likely the major source of polyamines in the blood and other extra-renal tissues in chickens.

Platinum (Pt) is one of the interesting catalysts in metal-assisted etching (metal-assisted chemical etching) of silicon (Si). The Pt-assisted etching induces not only the dissolution of Si under the Pt catalysts but also the formation of mesoporous layer on the Si surface away from them. In this work, we etched n-Si and p-Si by using patterned Pt films with a diameter of 5 μm and an interval of 50 μm. For both the cases, the Si surface under the Pt catalysts was selectively etched and macropores with a diameter of 5 μm were formed. The macropores formed on n-Si were deeper than those formed on p-Si. The mesoporous layer was observed only around the macropores on n-Si, while it was observed over the entire surface of p-Si. We also measured the open circuit potential of Si in the etching solution. The positive shift of potential of n-Si by the Pt deposition was smaller than that of p-Si except for the initial stage of etching, which can be explained by the polarization characteristics. We discussed the etching behavior of n-Si and p-Si on the basis of the results of structure observation and electrochemical measurements.

This study determined the oxidation of amino acids, glucose and fatty acid in enterocytes of developing chickens. Jejunal enterocytes were isolated from 0-, 7-, 21-, and 42-d-old broiler chickens, and incubated at 40°C for 30 min in Krebs-Henseleit bicarbonate buffer (pH 7.4) containing 5 mM D-glucose and one of the following: 0.5-5 mM L-[U-14C]glutamate, 0.5-5 mM L-[U-14C]glutamine, 0.5-5 mM L-[U-14C]aspartate, 0.5-5 mM L-[U-14C]alanine, 0.5-2 mM [U-14C]palmitate, D-[U-14C]glucose, 0.5-5 mM [U-14C]propionate, and 0.5-5 mM [1-14C]butyrate. 14CO2 produced from each 14C-labeled substrate was collected for determination of radioactivity. Among all the substrates studied, glutamate had the greatest rate of oxidation in enterocytes from 0- to 42-d-old chickens. Glutamate transaminases, rather than glutamate dehydrogenase, may be primarily responsible for initiating glutamate degradation. Rates of amino acid and fatty acid oxidation by cells increased (P0.05) with increasing their extracellular concentrations from 0.5 to 5 mM. Rates of glutamate and glucose oxidation in enterocytes decreased (P0.05) with increasing age, and rates of glutamine, aspartate, propionate, and butyrate oxidation were lower (P0.05) in 42-d-old chickens than in 0-d-old chickens. By contrast, oxidation of palmitate at 2 mM increased (P0.05) by 118% in cells from 42-d-old chickens, compared with 0-d-old chickens. Compared with glutamate, oxidation of glutamine, aspartate, alanine, propionate, butyrate, and palmitate was limited in cells from all age groups of chickens. Collectively, these results indicate that glutamate is the major metabolic fuel in enterocytes of 0- to 42-d-old chickens.Glucose and fatty acids have long been regarded as the primary sources of energy for the absorptive epithelial cells (enterocytes) of the avian small intestine. However, little is known about the use of amino acids for ATP production in these cells. Based on studies with mammalian enterocytes, we hypothesize that aspartate, glutamate, and glutamine provide the bulk of energy for the enterocytes of post-hatching developing chickens. To test this hypothesis, we isolated jejunal enterocytes from 0-, 7-, 21-, and 42-d-old male broiler chickens and performed metabolic studies. Our results indicated that: (1) glutamate (an amino acid) was the major energy source for the enterocytes of post-hatching chickens, (2) the biological oxidation of other amino acids (glutamine, aspartate, and alanine) was limited in chicken enterocytes, (3) glucose was the second most important metabolic fuel in chicken enterocytes, and (4) chicken enterocytes had a limited ability to degrade fatty acids but oxidized more long-chain fatty acids than short-chain fatty acids. We conclude that glutamate is the major source of energy in the enterocytes of post-hatching developing chickens.

Metal-assisted etching is a promising technique for microfabrication of semiconductors. In this method, porous silicon (Si) can be produced with a very simple procedure, and various nanostructures can be designed by changing the catalyst patterns. The kind of metal catalysts is one of the key factors to control the porous structure. In this work, we performed the etching of n-type Si (100) in a hydrofluoric acid solution containing hydrogen peroxide in the dark using silver, gold, and platinum particles electrolessly deposited at a constant coverage, and demonstrated the difference in the porous structures obtained for the different kind of metal catalysts. By comparing the mass loss of substrates with the depth of pores formed under the metal particles, we found that general corrosion occurred on the top-surface of the Si substrate around the metal particles even under the dark condition. The general corrosion depended on the metal species and it was explained by the formation and dissolution of a mesoporous layer. The kind of metal catalysts influences the dissolution of the Si surface not only under the metal catalysts but also between them.

Electroless deposition of metal particles on silicon (Si) followed by the metal-assisted etching (metal-assisted chemical etching) is a simple way to fabricate Si nanostructures. A composite porous structure consisting of straight macropores and a mesoporous layer can be created by platinum (Pt)-particle-assisted etching. In this work, we studied the composite structure formation on a highly-doped p-Si (ca. 5 × 1018 cm−3) in comparison with a moderately-doped p-Si (ca. 1 × 1015 cm−3). The composite structure drastically changed: the ratio of mesoporous layer thickness to macropore depth increased to 1.1 from 0.16 by using the highly-doped Si instead of the moderately-doped Si. The open circuit potential of Si in the etching solution shifted to the positive direction by the Pt deposition. The potential shift of highly-doped Si was smaller than that of moderately-doped Si, which can be explained by the polarization characteristics. We calculated the band bending in Si by using a device simulator that reproduced the conditions of Pt-particle-assisted etching. The results indicated that, in the case of highly-doped Si, the consumption rate of positive holes at the Si surface away from the Pt particles increases due to the tunneling effect, which is consistent with the thick mesoporous layer formation.

Sarcopenia is the decline in skeletal muscle mass, strength, and functions, which decreases the quality of life in elderly people. This study investigated the suppressive effect of turmeric (Curcuma longa) extract (TE) on muscle atrophy in dexamethasone (DEX)-treated mice and C2C12 myotubes. DEX treatment significantly decreased the muscle weight and significantly increased Fbxo32 and Murf1 expression in mice, and these changes were suppressed by the supplementation of an AIN-93 based diet with 2% TE. A similar pattern was observed in FBXO32 and MuRF1 protein expression. In C2C12 myotubes, DEX treatment significantly increased FBXO32 and MuRF1 gene and protein expression, and these increases were significantly suppressed by TE supplementation at a concentration of 200 µg/mL. Furthermore, one of the five TE fractions, which were separated by high-performance liquid chromatography had a similar effect with TE supplementation. The present study proposes the suppressive effect of turmeric on sarcopenia.

Polyamines (putrescine, spermidine, and spermine) are synthesized primarily from ornithine via ornithine decarboxylase (ODC) in mammals. Although avian tissues contain ODC activity, little is known about intracellular sources of ornithine for their polyamine synthesis. This study tested the hypothesis that arginase and proline oxidase contribute to polyamine synthesis in chickens. Kidney, jejunum, leg muscle, and liver from 0-, 7-, 14- and 21-day-old broiler chickens were assayed for the activities of arginase, proline oxidase (POX), ornithine aminotransferase (OAT), and ornithine decarboxylase (ODC). Kidney slices were also used to determine

L-glutamine (Gln) is the most abundant amino acid (AA) in the plasma and skeletal muscle of poultry, and L-glutamate (Glu) is among the most abundant AAs in the whole bodies of all avian tissues. During the first-pass through the small intestine into the portal circulation, dietary Glu is extensively oxidized to CO

While it has been hypothesized that brown adipocytes responsible for mammalian thermogenesis are absent in birds, the existence of beige fat has yet to be studied directly. The present study tests the hypothesis that beige fat emerges in birds as a mechanism of physiological adaptation to cold environments. Subcutaneous neck adipose tissue from cold-acclimated or triiodothyronine (T3)-treated chickens exhibited increases in the expression of avian uncoupling protein (avUCP, an ortholog of mammalian UCP2 and UCP3) gene and some known mammalian beige adipocyte-specific markers. Morphological characteristics of white adipose tissues of treated chickens showed increased numbers of both small and larger clusters of multilocular fat cells within the tissues. Increases in protein levels of avUCP and mitochondrial marker protein, voltage-dependent anion channel, and immunohistochemical analysis for subcutaneous neck fat revealed the presence of potentially thermogenic mitochondria-rich cells. This is the first evidence that the capacity for thermogenesis may be acquired by differentiating adipose tissue into beige-like fat for maintaining temperature homeostasis in the subcutaneous fat ‘neck warmer’ in chickens exposed to a cold environment. Supplementary Information The online version contains supplementary material available at 10.1007/s00726-021-02953-5.

L-glutamine (Gln) is the most abundant amino acid (AA) in the plasma and skeletal muscle of poultry, and L-glutamate (Glu) is among the most abundant AAs in the whole bodies of all avian tissues. During the first-pass through the small intestine into the portal circulation, dietary Glu is extensively oxidized to CO2, but dietary Gln undergoes limited catabolism in birds. Their extra-intestinal tissues (e.g., skeletal muscle, kidneys, and lymphoid organs) have a high capacity to degrade Gln. To maintain Glu and Gln homeostasis in the body, they are actively synthesized from branched-chain AAs (abundant AAs in both plant and animal proteins) and glucose via interorgan metabolism involving primarily the skeletal muscle, heart, adipose tissue, and brain. In addition, ammonia (produced from the general catabolism of AAs) and α-ketoglutarate (α-KG, derived primarily from glucose) serve as substrates for the synthesis of Glu and Gln in avian tissues, particularly the liver. Over the past 20 years, there has been growing interest in Glu and Gln metabolism in the chicken, which is an agriculturally important species and also a useful model for studying some aspects of human physiology and diseases. Increasing evidence shows that the adequate supply of dietary Glu and Gln is crucial for the optimum growth, anti-oxidative responses, productivity, and health of chickens, ducklings, turkeys, and laying fowl, particularly under stress conditions. Like mammals, poultry have dietary requirements for both Glu and Gln. Based on feed intake, tissue integrity, growth performance, and health status, birds can tolerate up to 12% Glu and 3.5% Gln in diets (on the dry matter basis). Glu and Gln are quantitatively major nutrients for chickens and other avian species to support their maximum growth, production, and feed efficiency, as well as their optimum health and well-being.

Several genome-wide association studies (GWASs) have reported the association between genetic variants and the habitual consumption of foods and drinks, however, no association data are available regarding the consumption of black tea. The present study aimed to identify genetic variants associated with black tea consumption in 12,258 Japanese participants. Data on black tea consumption were collected by a self-administered questionnaire, and genotype data were obtained from a single nucleotide polymorphism array. In the discovery GWAS, two loci met suggestive significance (p &lt, 1.0 &times, 10&minus, 6). Three genetic variants (rs2074356, rs144504271, and rs12231737) at 12q24 locus were also significantly associated with black tea consumption in the replication stage (p &lt, 0.05) and during the meta-analysis (p &lt, 5.0 &times, 8). The association of rs2074356 with black tea consumption was slightly attenuated by the additional adjustment for alcohol drinking frequency. In conclusion, genetic variants at the 12q24 locus were associated with black tea consumption in Japanese populations, and the association is at least partly mediated by alcohol drinking frequency.

Excess glucocorticoid secretion induces oxidative damage and muscle proteolysis and modulates glucose and lipid metabolism. It is known that the high-temperature (HT) treatment enhances corticosterone (CORT) secretion, muscle proteolysis, and mitochondrial reactive oxygen species (mtROS) generation in chickens. The present study investigated the co-effects of CORT on proteolysis and mtROS production, together with glucose, fatty acid, and amino acid metabolism in HT-treated cells. Myoblast cells were isolated from the major pectoralis muscle of five 0- or 1-day-old neonatal chicks and were precultured at 37°C/CO2 conditions for 48 h to reach subconfluent (80%-90%) conditions. Cells were then reseeded onto a 6- or 24-well microplate for the subsequent measurement, followed by the culture under a control temperature (37°C, control) or HT (41°C) conditions for 1 or 6 h. The HT-treated cells were cocultured with physiologically relevant concentrations of CORT (20 ng/mL in dimethyl sulfoxide). The HT treatment decreased cellular protein content (P < 0.05) and increased atrogin-1 mRNA levels and mtROS generation levels compared to the control group (P < 0.05), whereas HT/CORT co-treatment did not induce changes in either parameter. The mRNA level of glucose transporter-1 was decreased in HT-treated cells compared to that in normal cells (P < 0.05), and the decrease was increased in the CORT co-treatment (P < 0.05). While HT treatment did not alter pyruvate dehydrogenase kinase-4 mRNA level, the level was increased in the CORT co-treatment compared to the control and HT-treated cells (P < 0.05). Neither HT nor HT/CORT treatments altered the mRNA levels of fatty acid oxidation-related factors, carnitine palmitoyl transferase-1, and cluster of differentiation 36. The study conducted a metabolic analysis using gas chromatography-mass spectrometry. The results showed that HT/CORT-treated cells had decreased intracellular citrate and α-ketoglutarate levels (P < 0.05) and increased extracellular alanine and amino acid that have gluconeogenic properties, as well as increased aspartate, isoleucine, serine, methionine, and threonine levels (P < 0.05) compared to HT-treated cells. These results suggest that CORT may not affect proteolysis and mtROS production but can suppress pyruvate oxidation and promote alanine production in HT-treated chickens.

Obesity is a major global lifestyle disorder associated with gut microbiota. The health benefits of eggshell membrane (ESM) have been shown in previous reports, particularly as regards gut microbiota composition. Here, we investigated whether ESM improves lipid metabolism and alters gut microbiota in high‐fat diet‐fed mice. A total of 20 C57BL/6J mice aged 6 weeks were given either a control diet (CON), high‐fat diet (HFD), or high‐fat diet + 8% ESM powder (HESM) for 20 weeks. ESM supplementation in HFD‐fed mice reduced plasma triglycerides (TG) and liver total cholesterol (TC) and upregulated the expression of lipid metabolism genes carnitine palmitoyltransferase 1A and suppressor of cytokine signaling 2. Microbiota analysis showed increased relative abundance of the anti‐obesity bacterium, Lactobacillus reuteri, at 4, 12, and 16 weeks and reduced the abundance of inflammation‐related Blautia hydrogenotrophica, Roseburia faecis, and Ruminococcus callidus at 12 and 20 weeks. ESM‐supplemented mice had increased cecal isobutyrate, negatively correlated with B. hydrogenotrophica and Parabacteroides goldsteinii abundance. The results indicate that ESM supplementation in HFD‐fed mice reduced plasma TG and liver TC, possibly through alteration of lipid metabolism gene expression and gut microbiota composition, suggesting that ESM may be effective in obesity management., Eggshell membrane (ESM) supplementation for 20 weeks in HFD‐fed mice reduced plasma triglycerides (TG) and liver total cholesterol (TC). The upregulated gene expression of lipid metabolism genes carnitine palmitoyltransferase 1A and suppressor of cytokine signaling 2 suggest that lipid metabolism might be accelerated to increase TG production and clearance rates, as well as increase liver cholesterol excretion. Microbiota analysis showed increased relative abundance of the anti‐obesity bacterium and reduced abundance of inflammation‐related bacterium, which were negatively correlated with cecal isobutyrate levels.

Metal-assisted etching is a promising technique for microfabrication of semiconductors. In this method, porous silicon (Si) can be produced with a very simple procedure, and various nanostructures can be designed by changing the catalyst patterns. The kind of metal catalysts is one of the key factors to control the porous structure. In this work, we performed the etching of

1. Oleuropein (Ole) is a major phenolic compound in Olea europaea, with anti-oxidative, anti-obesity, and anti-inflammatory properties. To explore the effect of Ole on the physiology and metabolism...

Skin aging is one of the hallmarks of the aging process that causes physiological and morphological changes. Recently, several nutritional studies were conducted to delay or suppress the aging process. This study investigated whether nutritional supplementation of the eggshell membrane (ESM) has a beneficial effect on maintaining skin health and improving the skin aging process in vitro using neonatal normal human epidermal keratinocytes (NHEK-Neo) and in vivo using interleukin-10 knockout (IL-10 KO) mice. In NHEK-Neo cells, 1 mg/mL of enzymatically hydrolyzed ESM (eESM) upregulated the expression of keratinocyte differentiation markers, including keratin 1, filaggrin and involucrin, and changed the keratinocyte morphology. In IL-10 KO mice, oral supplementation of 8% powdered-ESM (pESM) upregulated the expression of growth factors, including transforming growth factor β1, platelet-derived growth factor-β and connective tissue growth factor, and suppressed skin thinning. Furthermore, voltage-gated calcium channel, transient receptor potential cation channel subfamily V members were upregulated by eESM treatment in NHEK-Neo cells and pESM supplementation in IL-10 KO mice. Collectively, these data suggest that ESM has an important role in improving skin health and aging, possibly via upregulating calcium signaling.

Heat stress (HS) induces muscle protein degradation as well as production of mitochondrial reactive oxygen species (ROS). In the present study, to improve our understanding of how protein degradation is induced by HS treatment in birds, a time course analysis of changes in the circulating levels of glucocorticoid and N(τ)-methylhistidine, muscle proteolysis-related gene expression, and mitochondrial ROS generation, was conducted. At 25 days of age, chickens were exposed to HS conditions (33 °C) for 0, 0.5, 1 or 3 days. While no alteration in plasma N(τ)-methylhistidine concentration relative to that of the control group was observed in the 0.5 day HS group, the concentration was significantly higher in the 3-d HS treatment group. Plasma corticosterone concentrations increased in response to 0.5-d HS treatment, but subsequently returned to near-normal values. HS treatment for 0.5 days did not change the levels of μ-calpain, cathepsin B, or proteasome C2 subunit mRNA, but increased the levels of mRNA encoding atrogin-1 (P

It has been shown that oleuropein, a phenolic compound in the fruit and leaves of the olive tree (Olea europaea) induces mammalian uncoupling protein 1 (UCP1) expression via an increased secretion of noradrenaline and adrenaline. This study investigated the effects of oleuropein on avian UCP (avUCP) expression as well as genes related to mitochondrial oxidative phosphorylation and biogenesis in cultured avian muscle cells, together with reactive oxygen species generation. Oleuropein induced avUCP as well as peroxisome proliferator-activated receptor γ coactivator-1α (PGC-1α), nuclear respiratory factor-1 (NRF1), mitochondrial transcription factor A (TFAM) and ATP5a1 (a component of mitochondrial adenosine triphosphate synthase) gene expression and cytochrome c oxidase activity, indicating the induction of mitochondrial biogenesis. Sirtuin-1 (SIRT1) gene expression was also up-regulated by this compound, which could contribute to an increase in PGC-1α activity. Oleuropein suppressed the level of superoxide generation per mitochondrion, possibly via the up-regulation of avUCP and manganese superoxide dismutase (MnSOD) expression. Based on these findings, this study is the first to show that oleuropein may induce avUCP expression in avian muscle cells independent of the catecholamines, in which PGC-1α may be involved.

Heat stress is a major factor inducing oxidative disturbance in cells. In the present study, we investigated the mechanism of overproduction of reactive oxygen species (ROS) in cultured avian muscle cells in response to heat stress, and also focused attention on the interaction of mitochondrial superoxide anions with altered NADPH oxidase (NOX), superoxide dismutase (SOD) and heme oxygenase-1 (HO-1) mRNA levels in heat-stressed cells. Exposure of cells to heat stress conditions (41°C, 6h) resulted in increased mitochondrial superoxide and intracellular ROS levels, and increased carbonyl protein content as compared with that of normal cells (37°C). The mitochondrial uncoupler 2,4-dinitrophenol lowered intracellular ROS levels in heat-stressed cells. Heat stress increased NOX4 mRNA and decreased HO-1 mRNA levels, while SOD1 and SOD2 mRNA levels remained relatively stable in heat-stressed cells. Addition of the superoxide scavenger 4-hydroxy TEMPO to the culture medium of heat-stressed cells restored mitochondrial superoxide and intracellular ROS levels as well as NOX4 and HO-1 mRNA levels to near-normal values. We suggest that mitochondrial superoxide production could play an influential role in augmenting oxidative damage to avian muscle cells, possibly via the up-regulation of NOX4 and down-regulation of HO-1 in heat-stressed avian muscle cells.

It has been shown that oleuropein, a phenolic compound in the fruit and leaves of the olive tree (Olea europaea) induces mammalian uncoupling protein 1 (UCP1) expression via an increased secretion of noradrenaline and adrenaline. This study investigated the effects of oleuropein on avian UCP (avUCP) expression as well as genes related to mitochondrial oxidative phosphorylation and biogenesis in cultured avian muscle cells, together with reactive oxygen species generation. Oleuropein induced avUCP as well as peroxisome proliferator-activated receptor γ coactivator-1α (PGC-1α), nuclear respiratory factor-1 (NRF1), mitochondrial transcription factor A (TFAM) and ATP5a1 (a component of mitochondrial adenosine triphosphate synthase) gene expression and cytochrome c oxidase activity, indicating the induction of mitochondrial biogenesis. Sirtuin-1 (SIRT1) gene expression was also up-regulated by this compound, which could contribute to an increase in PGC-1α activity. Oleuropein suppressed the level of superoxide generation per mitochondrion, possibly via the up-regulation of avUCP and manganese superoxide dismutase (MnSOD) expression. Based on these findings, this study is the first to show that oleuropein may induce avUCP expression in avian muscle cells independent of the catecholamines, in which PGC-1α may be involved.