• BMB Reports
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• v.55 no.4
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• pp.161-165
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• 2022

The mechanistic target of rapamycin (mTOR) regulates numerous extracellular and intracellular signals involved in the maintenance of cellular homeostasis and cell growth. mTOR also functions as an endogenous inhibitor of autophagy. Under nutrient-rich conditions, mTOR complex 1 (mTORC1) phosphorylates the ULK1 complex, preventing its activation and subsequent autophagosome formation, while inhibition of mTORC1 using either rapamycin or nutrient deprivation induces autophagy. Autophagy and proteasomal proteolysis provide amino acids necessary for protein translation. Although the connection between mTORC1 and autophagy is well characterized, the association of mTORC1 inhibition with proteasome biogenesis and activity has not been fully elucidated yet. Proteasomes are long-lived cellular organelles. Their spatiotemporal rather than homeostatic regulation could be another adaptive cellular mechanism to respond to starvation. Here, we reviewed several published reports and the latest research from our group to examine the connection between mTORC1 and proteasome. We have also investigated and described the effect of mTORC1 inhibition on proteasome activity using purified proteasomes. Since mTORC1 inhibitors are currently evaluated as treatments for several human diseases, a better understanding of the link between mTORC1 activity and proteasome function is of utmost importance.

• Asian-Australasian Journal of Animal Sciences
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• v.24 no.1
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• pp.88-95
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• 2011

An experiment was conducted to determine the standardized ileal digestibility (SID) of amino acids (AA) in four sources of full-fat soybeans (FFSB) and in one source of soybean meal (SBM). The FFSB had different concentrations of trypsin inhibitor units (TIU) and included two sources of conventional FFSB, and two sources of a soybean variety that was selected for a reduced concentration of the Kunitz trypsin inhibitor. The conventional FFSB was either low temperature-processed (LT-FFSB-CV; 37.7% CP, 35.4 TIU/mg) or high temperature-processed (HT-FFSB-CV; 40.5% CP, 4.4 TIU/mg). The low-Kunitz FFSB was also either low temperature-processed (LT-FFSB-LK; 36.2% CP, 23.5 TIU/mg) or high temperature-processed HT-FFSB-LK; (38.2% CP, 4.0 TIU/mg). The SBM contained 47.5% CP and 3.20 TIU/mg. Twelve weanling barrows (initial BW: $11.1{\pm}1.3\;kg$) were fitted with a T-cannula in the distal ileum. Pigs were allotted to a replicated $6{\times}6$ Latin square design with six diets and six periods per square. Five diets were prepared using each of the soybean sources as the only source of AA in the diet. An N-free diet was also included in the experiment to measure basal endogenous losses of AA. The two low temperature-processed FFSB had lower (p<0.05) AID and SID values for all indispensable AA than the two high temperature-processed FFSB and SBM. The SID values for all indispensible AA except Trp were greater (p<0.05) in LT-FFSB-LK than in LT-FFSB-CV, but the SID of AA in HT-FFSB-CV and HT-FFSB-LK were not different. The SID of AA in SBM were not different from the SID in HT-FFSB-CV and in HT-FFSB-LK. Results of this experiment show that a reduction of the TIU from 35.4 to 23.5 TIU/mg will improve the SID of AA, but this reduction is not sufficient to completely ameliorate the negative impact of trypsin inhibitors. Results also show that the SID of AA in high temperature-processed FFSB is similar to that in de-hulled SBM.

• Asian-Australasian Journal of Animal Sciences
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• v.27 no.5
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• pp.706-716
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• 2014

A new strategy of co-inoculating Bacillus subtilis MA139 with Streptococcus thermophilus and Saccharomyces cerevisiae was used to produce fermented soybean meal (FSBM). Three experiments were conducted to determine the concentration of digestible energy (DE) and metabolizable energy (ME) (Exp. 1), apparent ileal digestibility (AID) and standardized ileal digestibility (SID) of amino acids (AA) (Exp. 2), and feeding value (Exp. 3) of FSBM produced by this new strategy (NFSB) compared with soybean meal (SBM) and conventionally available FSBM (Suprotein). In Exp. 1, twenty-four barrows (initial body weight [BW] of $32.2{\pm}1.7kg$) were randomly allotted to 1 of 4 diets with 6 replicates per diet. A corn basal diet and 3 diets based on a mixture of corn and 1 of 3 soybean products listed above were formulated and the DE and ME contents were determined by the difference method. The results showed that there were no differences in DE and ME between SBM and either FSBM product (p>0.05). In Exp. 2, eight barrows (initial BW of $26.8{\pm}1.5kg$) were fitted with ileal T-cannulaes and used in a replicated $4{\times}4$ Latin square design. Three corn-starch-based diets were formulated using each of the 3 soybean products as the sole source of AA. A nitrogen-free diet was also formulated to measure endogenous losses of AA. The results showed that the SID of all AA except arginine and histidine was similar for NFSB and SBM (p>0.05), but Suprotein had greater (p<0.05) SID of most AA except lysine, aspartate, glycine and proline than NFSB. In Exp. 3, a total of 144 piglets (initial BW of $8.8{\pm}1.2$ kg) were blocked by weight and fed 1 of 4 diets including a control diet with 24% SBM as well as diets containing 6% and 12% NFSB or 12% Suprotein added at the expense of SBM. During d 15 to 28, replacing SBM with 6% NFSB significantly improved average daily gain (ADG) and average daily feed intake (ADFI) (p<0.05) for nursery piglets. During the overall experiment, ADG of piglets fed diets containing 6% NFSB was significantly greater (p<0.05) than that of piglets fed SBM. In conclusion, fermentation with the new strategy did not affect the energy content or the AID and the SID of AA in SBM. However, inclusion of 6% NFSB in diets fed to nursery piglets improved performance after weaning likely as a result of better nutritional status and reduced immunological challenge.

• Journal of Animal Science and Technology
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• v.62 no.3
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• pp.374-384
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• 2020

Two experiments were conducted to determine apparent metabolizable energy (AME), nitrogen-corrected AME (AMEn), and ileal digestible amino acid (AA) content of brown rice (BR) and to investigate the effect of dietary supplementation of BR on laying performance and egg quality of laying hens. In Exp. 1, 72 Hy-line Brown layers (49-week-old) were allocated to two treatments using a completely randomized block design, and each treatment included six cages per treatment and six hens per cage. A semi-purified diet was formulated to include BR as the sole source of AA and energy and an N-free diet was used to determine basal endogenous loss of AA. The hens were fed a commercial layer diet for adaptation to the experimental environment and diet for 7 days from d 0, and then fed experimental diets for 5 days from d 7. Excreta were collected from d 10 to 11 and ileal digesta were collected on d 12. On a dry matter (DM) basis, the AME and AMEn of BR was determined at 3,773 and 3,729 kcal/kg, respectively. The apparent ileal digestibility (AID) of BR ranged from 32.7% for Thr to 73.7% for Arg. The range of the standardized ileal digestibility (SID) value was between 79.4% for Met and 96.6% for Lys. In Exp. 2, 252 Hy-line Brown layers (44-week-old) were divided into four groups, comprising seven replicates of nine birds each and assigned to four experimental diets containing 0 (Control), 5%, 10%, or 15% BR for 5 weeks. The BR-containing diets were formulated to be equal in the content of AMEn and digestible AA to those of the diet without BR. No significant differences were observed in laying performances. Egg quality and blood profiles were not linearly or quadratically affected by dietary treatments. These results suggest that up to 15% BR can be included into layer feed without any adverse effects on laying performance and egg quality, if its energy and digestible AA values are well evaluated.

• Asian-Australasian Journal of Animal Sciences
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• v.27 no.7
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• pp.990-995
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• 2014

The objective of this experiment was to determine the concentration and digestibility of crude protein (CP) and amino acid (AA) in meat meal (MM), and to compare these values with the respective values in soybean meal (SBM). Six barrows (initial body weight = $66.9{\pm}3.8kg$) surgically fitted with a T-cannula at the distal ileum were allotted to a replicated $3{\times}3$ balanced Latin square design with 3 diets and 3 periods. Two experimental diets containing test ingredients as the sole source of AA were prepared to estimate the apparent ileal digestibility (AID) for CP and AA by the direct method. An N-free diet was also prepared to estimate basal endogenous losses of CP and AA. All experimental diets contained 5% chromic oxide as an indigestible index. Each period consisted of a 5-d adaptation period and a 2-d of ileal digesta collection period. Ileal digesta samples were collected from 0900 to 1700 on d 6 and 7 of each period. The concentrations of CP, Lys, Met, and Trp in MM and SBM were analyzed to be 64.1, 3.5, 1.1 and 0.6, and 45.6, 2.8, 0.8, and 0.3%, respectively. The AID of all AA except Gly in MM was less (p<0.05) than in SBM. The AID of Lys, Met, and Trp in MM was estimated to be 56.0, 71.7, and 47.1%, respectively. The SID of all AA in MM was less (p<0.05) than in SBM. The SID of Lys, Met, and Trp was 65.1, 79.2, and 78.5%, respectively. In conclusion, the CP and AA contents in MM were greater than those in SBM whereas the ileal digestibility of all AA in MM was less than in SBM.

• Molecules and Cells
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• v.28 no.6
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• pp.575-581
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• 2009

Ornithine decarboxylase (ODC) is a rate-limiting enzyme in the biosynthesis of polyamines, which are essential for cell growth, differentiation, and proliferation. This report presents the characterization of an ODC-encoding cDNA (SlitODC) isolated from a moth species, the tobacco cutworm, Spodoptera litura (Lepidoptera); its expression in a polyamine-deficient strain of yeast, S. cerevisiae; and the recovery in polyamine levels and proliferation rate with the introduction of the insect enzyme. SlitODC encodes 448 amino acid residues, 4 amino acids longer than B. mori ODC that has 71% identity, and has a longer C-terminus, consistent with B. mori ODC, than the reported dipteran enzymes. The null mutant yeast strain in the ODC gene, SPE1, showed remarkably depleted polyamine levels; in putrescine, spermidine, and spermine, the levels were > 7, > 1, and > 4%, respectively, of the levels in the wild-type strain. This consequently caused a significant arrest in cell proliferation of > 4% of the wild-type strain in polyamine-free media. The transformed strain, with the substituted SlitODC for the deleted endogenous ODC, grew and proliferated rapidly at even a higher rate than the wild-type strain. Furthermore, its polyamine content was significantly higher than even that in the wild-type strain as well as the spe1-null mutant, particularly with a very continuously enhanced putrescine level, reflecting no inhibition mechanism operating in the putrescine synthesis step by any corresponding insect ODC antizymes to SlitODC in this yeast system.

• Archives of Pharmacal Research
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• v.22 no.5
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• pp.464-473
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• 1999

Regulation of phosphcholine-hydrolyzing phosphatase (phosphocholine-phosphatase) activity, purified from bovine brain, was examined under physiological conditions. Various endogenous phosphomonoesters, which were utilized as substrate, inhibited the phosphocoline-phosphatase activity competitively (Ki 5.5-$82.0 {\mu}M$); among phosphomonoesters tested, there was a similar order of capability between the binding affinity of substrate and the inhibitory potency. In addition, phosphate ions also inhibited the phosphatase activity competitively with a Ki value of approximately $16{\mu}M$. Although leucine or theophylline inhibited the phosphatase activity at pH 9.0, their inhibitory action decreased greatly at pH 7.4. The pH-Km and pH-Vm profiles indicate that ionizable amino acids are involved in substrate binding as well as catalysis, alluding that the phosphatase activity may be highly dependent on the intracellular pH. Amino acid modification study supports the existence of tyrosine, arginine or lysine residue in the active site, and the participation of tyrosine residue in the catalytic action may e suggested positively for the susceptibility to the action of tetranitromethane or HOl-generator. Separately, the oxidative inactivation of phosphocholine-phosphatase activity was investigated. Of oxidants tested, HOONO, HOCl, HOl and $ascorbate/Cu^{2+}$ system were effective to inactivate the phosphatase activity. Noteworthy, a remarkable inativation was accomplished by $30{\mu}M$ HOCl in combination with 1 mM Kl. Inaddition, $Cu^{2+}(3{\mu}M) $in combination with ascorbate at concentrations as low as 0.1-0.3 mM reduced the phosphatase activity to a great extent. From these results, it is proposed that the phosphocholine-phosphatase activity may be regulated endogenously and susceptible to the various oxidant system in vivo.

• Asian-Australasian Journal of Animal Sciences
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• v.32 no.11
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• pp.1745-1752
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• 2019

Objective: The objective was to determine standardized ileal digestibility (SID) of amino acids (AA) in 11 plant protein sources fed to growing pigs. Methods: Eleven feed ingredients used were sesame meal, two sources of soybean meal (SBM) produced in the Republic of Korea, a source of SBM produced in India, high-protein distillers dried grains (HPDDG), perilla meal, canola meal, copra meal, corn germ meal, palm kernel expeller, and tapioca distillers dried grains (TDDG). Experimental diets were prepared to contain each test ingredient as a sole source of AA, and a nitrogen-free diet was also prepared to estimate the basal ileal endogenous losses of AA. Twelve barrows surgically fitted with T-cannulas at the distal ileum with an initial body weight of 29.0 kg (standard deviation = 3.0) were individually housed in metabolism crates equipped with a feeder and a nipple drinker. A $12{\times}9$ incomplete Latin square design was employed with 12 experimental diets, 12 animals, and 9 periods. After a 5-d adaptation period, ileal digesta were collected on d 6 and 7 in each experimental period. Results: Values for apparent ileal digestibility of most indispensable AA in three sources of SBM were greater compared with other test ingredients except HPDDG and canola meal (p<0.05). Pigs fed diets containing SBM sources had also greater SID of most indispensable AA compared with those fed diets containing other test ingredients (p<0.05) except for HPDDG and canola meal. There was no difference in the apparent ileal digestibility and SID of AA among sources of SBM. The TDDG had the least value for the SID of methionine among test ingredients (p<0.05). Conclusion: The SID of most AA in SBM, HPDDG, and canola meal were greater than those in sesame meal, perilla meal, copra meal, and TDDG.

• Journal of The Korean Society of Grassland and Forage Science
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• v.42 no.3
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• pp.195-200
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• 2022

Sulfur is an essential element in plants, including amino acids, vitamin synthesis, and acting as an antioxidant. However, the interaction between endogenous sulfur and proline synthesis has not been yet fully documented. White clover (Trifolium repens L.) is known as a species highly sensitive to sulfate supply. Therefore, this study aimed to elucidate the role of sulfur in regulating proline metabolism in relation to ammonia detoxification and hydrogen peroxide (H₂O₂) accumulation in white clover. The detached leaves of white clover were immersed in solution containing different concentration of sulfate (0, 10, 100, and 1000 mM MgSO₄). As MgSO₄ concentrations were increased, the concentration of H₂O₂ increased up to 2.5-fold compared to control, accompanied with H₂O₂ detection in leaves. Amino acid concentrations significantly increased only at higher levels (100 and 1000 mM MgSO₄). No significant difference was observed in protein concentration. Proline and ∆^1-pyrroline-5-carboxylate (P5C) concentrations slightly decreased at 10 and 100 mM MgSO₄ treatments, whereas it rapidly increased over 1.9-fold at 1000 mM MgSO₄ treatment. Ammonia concentrations gradually increased up to 8.6-fold. These results indicate that exogenous sulfur levels are closely related to H₂O₂ and ammonia synthesis but affect proline biosynthesis only at a higher level.

• Asian-Australasian Journal of Animal Sciences
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• v.26 no.5
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• pp.691-699
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• 2013

Three experiments were conducted to evaluate the metabolizable energy (ME) value, standardized ileal digestibility (SID) of amino acids (AA) of soybean meal (SBM), soy protein concentrate (SPC) and fermented soybean meal (FSBM), and the application of these products in early-weaned piglets. In Exp. 1, four barrows with initial body weight (BW) of $14.2{\pm}1.4$ kg were used in a $4{\times}4$ Latin square design. The diet 1 contained corn as the only energy source. The other three diets replaced 25% of corn in diet 1 with one of the three soybean products, and the digestable energy (DE) and ME contents were determined by difference. In Exp. 2, four barrows (initial BW of $18.2{\pm}1.5$ kg) were fitted with ileal T-cannulas and allotted to a $4{\times}4$ Latin square design. Three cornstarch-based diets were formulated using each of the soybean products as the sole source of AA. A nitrogen-free diet was also formulated to measure endogenous losses of AA. In Exp. 3, ninety six piglets (initial BW of $5.6{\pm}0.9$ kg) weaned at $21{\pm}2$ d were blocked by weight and assigned to one of three treatments for a 21-d growth performance study. The control diet was based on corn and SBM, the two treatments' diets contained either 10% SPC or FSBM and were formulated to same SID lysine to ME ratio of 3.6 g/Mcal. The results showed that the ME content of SPC was greater than SBM (p<0.05). The SID of most AA in SPC was greater than the SID of AA in SBM (p<0.05). For the essential AA, the SID of histidine, isoleucine, leucine, lysine and threonine in FSBM were greater than in SBM (p<0.05). Even though they were fed same SID lysine to ME ratio of 3.6 g/Mcal diets, pigs fed SPC and FSBM diets had greater weight gain, G:F (p<0.05) and better fecal score (p<0.05) than pigs fed SBM diet. In conclusion, SPC showed a higher ME content and SID of AA than the SBM. SID of some essential AA in FSBM was higher than SBM and was similar with SPC. But the lower antigenic proteins and anti-nutritional factors content in SPC and FSBM may be the main factors affecting the performance of early-weaned piglets rather than the increased ME content and SID of AA.