• Journal of Animal Science and Technology
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• v.64 no.3
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• pp.481-499
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• 2022

This study aims to determine the effects of D-methionine (D-Met) isomer and the methionine precursor 2-hydroxy-4-methylthiobutanoic acid i (HMBi) supplementation on milk protein synthesis on immortalized bovine mammary epithelial cell (MAC-T). MAC-T cells were seeded using 10-cm dishes and cultured in Dulbecco's modified Eagle's medium/F12 (DMEM/F12) basic medium. The basic medium of DMEM/F12 was replaced with the lactogenic DMEM/ F12 differentiation medium when 90% of MAC-T cells reached confluency. The best dosage at 0.6 mM of D-Met and HMBi and incubation time at 72 h were used uniformly for all treatments. Each treatment was replicated six times wherein treatments were randomly assigned in a 6-well plate. Cell, medium, and total protein were determined using a bicinchoninic acid protein assay kit. Genes, proteomics and metabolomics analyses were also done to determine the mechanism of the milk protein synthesis pathway. Data were analyzed by two-way analysis of variance (ANOVA) with supplement type and plate as fixed effects. The least significant difference test was used to evaluate the differences among treatments. The HMBi treatment group had the highest beta-casein and S6 kinase beta-1 (S6K1) mRNA gene expression levels. HMBi and D-Met treatments have higher gene expressions compared to the control group. In terms of medium protein content, HMBi had a higher medium protein quantity than the control although not significantly different from the D-Met group. HMBi supplementation stimulated the production of eukaryotic translation initiation factor 3 subunit protein essential for protein translation initiation resulting in higher medium protein synthesis in the HMBi group than in the control group. The protein pathway analysis results showed that the D-Met group stimulated fructose-galactose metabolism, glycolysis pathway, phosphoinositide 3 kinase, and pyruvate metabolism. The HMBi group stimulated the pentose phosphate and glycolysis pathways. Metabolite analysis revealed that the D-Met treatment group increased seven metabolites and decreased uridine monophosphate (UMP) production. HMBi supplementation increased the production of three metabolites and decreased UMP and N-acetyl-L-glutamate production. Taken together, D-Met and HMBi supplementation are effective in stimulating milk protein synthesis in MAC-T cells by genes, proteins, and metabolites stimulation linked to milk protein synthesis.

• Asian-Australasian Journal of Animal Sciences
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• v.14 no.4
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• pp.525-534
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• 2001

A total of 120 pigs were used to investigate the effects of yucca extracts on the growth performance, nutrient digestibility, nutrient excretion and carcass characteristics of finishing pigs fed different levels of dietary protein. Pigs were allotted into $2{\times}3$ factorial design by the supplementation of yucca extracts (YE, 0 and 120 mg/kg) and 3 levels of dietary protein (14, 16, 18% for early finisher and 12, 14, 16% for late finisher for low, medium and high protein diet, respectively). During the early finishing period (51~76 kg BW), no significant difference was found in growth performance regardless of the YE supplementation or dietary protein levels. Growth performance of late finishing pigs (76~101 kg BW) was also not significantly different among treatments. However, ADG of pigs fed YE diet was significantly improved (p<0.05) regardless of the dietary protein levels. For the overall period (51~101 kg BW), although adding YE to the diet and elevating the protein level showed better ADG, there were no significant differences on growth performance among treatments. Early finishers showed significantly higher crude protein, crude ash and crude fat digestibilities when they were fed diets supplemented with YE. Digestibilities of amino acids were not affected by YE. Late finishers did not show any significant differences in proximate nutrient digestibilities regardless of YE supplementation or dietary protein levels. YE tended to slightly improve the CP digestibility, however no significant difference was found with increased dietary protein levels. There was no significant difference in amino acid digestibilities with YE supplementation or dietary CP levels during the late finishing period. Dry matter (DM) and nitrogen (N) excretion in feces did not show any significant difference among treatments. Early finishing pigs also did not respond to the inclusion of YE or dietary protein levels (p<0.05). Fecal N excretion of early finishing pigs seemed to be lowered in pigs fed YE. Pigs fed medium dietary protein diet tended to excrete a higher amount of N during the early finishing period, but not statistically different. A slight increase in fecal N excretion was found with the increased level of dietary protein during the late finishing period. For ammonia nitrogen excretion, although there was no significance, the NH3-N content tended to be increased by the increased dietary protein levels and with YE supplementation. The NH3-N content in manure increased by 24.5% with YE supplementation. There were no significant differences in carcass weight, backfat thickness, carcass grade and loin eye area among treatments. However, pigs fed non-YE with low protein diet showed a significantly (p<0.05) low carcass ratio among treatments and there was significant (p<0.05) difference between the YE-added treatment and non YE treatment in carcass ratio. As for the feed cost, the cost of feeding high level protein was higher than that of medium level protein by 5% and low level protein by 9% (p<0.05). Therefore, based on this study, it could be concluded that environmentally friendly agents might play a role to some extent in finishing pigs from the aspect of pollution control, and that more than 14 and 12% of dietary protein for early finishing and late finishing pigs respectively do not necessarily guarantee high growth performance.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.8
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• pp.1080-1087
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• 2004

A trial with twenty-four newly weaned Awassi lambs (initial body weight=21.5$\pm$0.8 kg) was conducted using a 3$\times$2 factorial design to study the effect of feeding three sources of protein supplements (soybean meal (SBM), sunflower seed meal (SSM), and cottonseed meal (CSM)), either untreated or formaldehyde-treated on the growth performance and carcass traits of Awassi lambs. Lambs were randomly assigned to one of the six diets (4 lambs/treatment diet) and were individually fed for a period of 107 days. Experimental diets were isonitrogenous and isocaloric. Final live weight and average daily gain (ADG) were affected by both source of protein and formaldehyde treatment (undegradable protein). Lambs fed untreated diets had better (p<0.01) daily gain compared to those fed formaldehyde-treated diets. Similarly total feed intake per animal was significantly (p<0.05) affected by protein source and formaldehyde treatment. Formaldehyde treatment caused a significant decrease (p<0.01) in feed intake compared to lambs fed untreated diets. Feed requirement per unit of gain was not affected by formaldehyde treatment during all periods of the experiment except for the second period (the second 28 day period), whereby untreated SBM, SSM and CSM had better feed conversion ratio (FCR) than the treated groups. Source of protein had a moderate effect (p<0.10) on FCR but had a significant effect (p<0.05) on hot and cold carcass weight, digestive tract empty weight and liver weight, with lambs fed SBM having higher values than lambs fed SSM and CSM diets. Supplementation with undegradable protein had a significant effect (p<0.05) on dressing-out percentage (p<0.05), final live weight, and hot and cold carcass weight (p<0.01). The lower values pertain to lambs fed treated diets compared to lambs fed untreated diets. In general, there were no significant differences among all carcass linear dimensions, carcass cut weights and dissected loin tissue weights for both treatments (protein source and formaldehyde treatment). Supplementation with undegradable protein but not the source of protein resulted in significantly higher dissected leg total bone weight (p<0.05), tibia and femur weight (p<0.05), and femur length (p<0.01) at the same carcass weight. Results suggest that the treatment of SBM, SSM and CSM with formaldehyde did not improve efficiency of feed utilization, lamb performance or carcass traits and that the SBM diet resulted in an increase in lamb performance compared to other experimental diets.

• Journal of Nutrition and Health
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• v.29 no.7
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• pp.788-805
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• 1996

This study was performed to investigate effects of examination-stress and protein supplementation on nitrogen metabolism and blood protein levels of Korean college students. Experiment was conducted at the beginning of a academic term and during midterm examination. During midterm examination, subjects were classified into two groups randomly : protein supplemental group(male n=6, female n=10) and placebo group(male n=4, female n=9). Protein capsules(2g/day) above 10% of indispensible amino acids requirement estimates were given to supplemental group for 10 days. At the begining of the term, male students(n=12) ingested 223.15mgN/kg/d, excreted 20.7mgN/kg/d in feces, and excreted 94.31mgN/kg/d in urine. Their apparent protein protein digestibility was 90.72%, true N balance was +100.11mgN/kg/d, and the mean maintenance N requirement of mixed Korena diet calculated was 112.13mgN/kg/d. Female students(n=19) ingested 171.44mgN/kg/d, excreted 22.13mgN/kg/d in feces, and excreted 122.92mgN/kg/d in urine. Their apparent protein digestibility was 86.76%, true N blance was + 18.39mgN/kg/d, and the mean maintenance N requirement calculated was 135.31mgN/kg/d. Blood levels of serum total protein, albumin, and BUN were within normal range. During midterm examination, fecal and urinary N excretions of female subjects(n=19) were increased, especially urea N markedly, and urea N/creatinine N ratio was augumented significantly. Apparent protein digestibility of male subjects(n=10) was decreased. Examination-stress showed 8.05mgN/kg/d (7.2%) increase of mean maintenance N requirement in male and 8.55mgN/kg/d(6.3%) increase in female students in comparison with that of the beginning of the term. Serum total protein and albumin levels showed no significant change, but serum transferrin level of female were decreased significantly. During midterm examination, females supplemented with protein capsules(2g/d)had no significant increase in fecal and urinary N excretions.

• Korean journal of food and cookery science
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• v.13 no.5
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• pp.627-634
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• 1997

The purpose of this study is to examine the effect of protein content on the physicochemical properties, gelatinized characteristics and textural properties of cowpea precipitate gels stored for 24 hrs and 48 hrs at room temperature. The contents of protein, total fat, and ash ranged from 0.35%∼1.38%, 0.54%∼0.64%, and 0.21%∼0.25%, respectively. The X-ray diffraction patterns were all Ca-type, showing no difference according to the protein content. Protein content did not make any difference in the blue values of cowpea precipitate. The blue value of cowpea precipitate powder as protein content was decreased. The water-binding capacity of cowpea precipitate powder increased as the protein content increased. Swelling power and solubility of cowpea precipitate powder increased as protein content decreased. The transmittance of cowpea precipitate powder was not different according to the protein content. The initial pasting temperature of cowpea precipitate powder by differential scanning calorimetry (DSC) and rapid visco analyser (RVA) showed no differences according to the protein content. In sensory evaluation, the color and clarity of cowpea precipitate gels stored for 24 hrs and 48 hrs at room temperature as the protein content increased, and the hardness, cohesiveness, springiness, acceptability were greater when the gels were stored for 48 hrs. Instrumental analyses using a rheometer showed that the hardness, gumminess, and chewiness of cowpea precipitate gels stored for 24 hrs, which was increased as the high protein content increased. For the gels stored for 48 hrs, all other factors are significantly different except cohesiveness as the protein content increased.

• Asian-Australasian Journal of Animal Sciences
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• v.11 no.3
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• pp.311-317
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• 1998

A total of 1033 Brown Swiss and 610 Canadienne cows were phenotyped for the genetic variants ${\alpha}_{s1}$-casein, ${\beta}$-casein, ${\kappa}$-casein, ${\beta}$-lactoglobulin and ${\alpha}$-lactalbumin. In Brown Swiss, frequency distributions were: 97.3% B and 2.7% C variant of ${\alpha}_{s1}$-casein; 31.6% $A^1$, 51.8% $A^2$, 0.5% $A^3$ and 16.1% B variant of ${\beta}$-casein; 70.4% A, 29.3% B, and 0.3% C variant of ${\kappa}$-casein; 41.7% A and 58.3% B variant of ${\beta}$-lactoglobulin; and 100% B variant of ${\alpha}$-lactalbumin. Corresponding frequencies in Canadienne for those five milk proteins were: 98.6 and 1.4%;58.5, 33.5, 0.08 and 7.9%; 78.8, 21.1 and 0.1%, 42.4 and 57.6%; and 100%. Analysis of variance by least squares showed possible association between milk protein phenotypes and some lactational production traits. There were no significant association of phenotypes of ${\alpha}_{s1}$-casein, ${\beta}$-casein and ${\beta}$-lactoglobulin with milk yield, fat yield, protein yield, fat percentage and protein percentage in both breeds during the three lactations. In the Brown Swiss, ${\kappa}$-casein phenotype was associated with 305-day fat yield and protein yield during the first lactation. ${\kappa}$-Casein AB was associated with higher milk, fat and protein yield during the second lactation. During the third lactation, ${\beta}$-lactoglobulin AA in Canadienne cows was associated with higher protein content in the milk (3.70%) when compared to phenotypes AB (3.54%) and BB (3.64%).

• Journal of KIISE:Software and Applications
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• v.36 no.10
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• pp.851-860
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• 2009

Recently, among the computational methods of protein-protein interaction prediction, vast amounts of domain based methods originated from domain-domain relation consideration have been developed. However, it is true that multi domains collaboration is avowedly ignored because of computational complexity. In this paper, we implemented a protein interaction prediction system based the Interaction Significance matrix, which quantified an influence of domain combination pair on a protein interaction. Unlike conventional domain combination methods, IS matrix contains weighted domain combinations and domain combination pair power, which mean possibilities of domain collaboration and being the main body on a protein interaction. About 63% of sensitivity and 94% of specificity were measured when we use interaction data from DIP, IntAct and Pfam-A as a domain database. In addition, prediction accuracy gradually increased by growth of learning set size, The prediction software and learning data are currently available on the web site.

• Proceedings of the Korean Vacuum Society Conference
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• 2015.08a
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• pp.230.1-230.1
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• 2015

One of the major problems of biological ToF-SIMS imaging is the lack of protein and peptide imaging. Most of biological story telling is mianly based on proteins. The biological implication of lipid ToF-SIMS imaging would be much higher if protein imaging is provided together. Utilizing high secondary ion yields of metals, proteins can be ToF-SIMS imaged with nanoparticle tagged proteins. Nanoparticles such as Fe3O4, SiO2, PbS were used for imaing NeuN, MCH, Orexin A, ${\alpha}$ synucline, TH(Tryosine Hydroxylase) in mouse tissues with a spatial resolution of ${\sim}2{\mu}m$ using a TOF-SIMS. Lipids and neurotransmitters images obtained simultaneously with protein images were overlayed for more deeper understanding of neurobiology, which is not allowed by any other bioimaging technqiues. The protein images from TOF-SIMS were compared with confocal fluorescence microscopy and NanoSIMS images. A new sample preparation method for imaging single cell membranes in a tissue using the vibrotome technique to prepare a tissue slice without any fixation and freeze drying will be also presented briefly for Hippocampus and Hypothalamus tissues.

• BMB Reports
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• v.28 no.2
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• pp.149-154
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• 1995

Protein methylase II (S-adenosyl-L-methionine : protein O-methyl-transferase, EC 2.1.1.24; PM II) was purified approximately 1250-fold from porcine testis by fractional precipitation and DEAE-cellulose chromatography, followed by gel filtration on a Sephadex G-75 column and HPLC on a Protein Pak 125 column. The molecular weight of the enzyme was estimated to be 33,000 daltons by SDS-PAGE, which agreed with the value determined by gel filtration. Isoelectric focusing of purified PM II showed a single protein species with an isoelectric point of 6.2. The optimum pH for the reaction was 6.0. The $K_m$ value of the enzyme was $1{\times}10^{-5}M$ with a $V_{max}$ value of 769 pmol/min/mg of enzyme. S-adenosyl-L-homocysteine is a competitive inhibitor of PM II with a $K_i$ value of $1.38{\times}10^{-6}M$.

• Interdisciplinary Bio Central
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• v.3 no.2
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• pp.8.1-8.6
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• 2011

Background: Thanks to the development of the mathematical/statistical reverse engineering and the high-throughput measuring biotechnology, lots of biologically meaningful genegene interaction networks have been revealed. Steady-state analysis of these systems provides an important clue to understand and to predict the systematic behaviours of the biological system. However, modeling such a complex and large-scale system is one of the challenging difficulties in systems biology. Results: We introduce a new stochastic modeling approach that can describe gene regulatory mechanisms by dividing two (DNA and protein) layers. Simple queuing system is employed to explain the DNA layer and the protein layer is modeled using G-networks which enable us to account for the post-translational protein interactions. Our method is applied to a transcription repression system and an active protein degradation system. The steady-state results suggest that the active protein degradation system is more sensitive but the transcription repression system might be more reliable than the transcription repression system. Conclusions: Our two layer stochastic model successfully describes the long-run behaviour of gene regulatory networks which consist of various mRNA/protein processes. The analytic solution of the G-networks enables us to extend our model to a large-scale system. A more reliable modeling approach could be achieved by cooperating with a real experimental study in synthetic biology.