• The Korean Journal of Physiology and Pharmacology
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• v.3 no.4
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• pp.447-454
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• 1999

The production of nitric oxide (NO) and the expression of inducible nitric oxide synthase (iNOS) are known to be modulated by a variety of factors. Recent study showed that endotoxin-induced NO synthesis and iNOS expression were greatly enhanced by elevation of extracellular glucose concentration in murine macrophages. Although this was suggested to be due to the activation of protein kinase C (PKC) via sorbitol pathway, there was lack of evidence for this speculation. This study was performed to delineate the underlying intracellular mechanisms of glucose-enhancing effect on endotoxin-induced NO production in Raw264.7 macrophages. The levels of NO release induced by lipopolysaccharide (LPS) significantly increased by the treatment of glucose in a concentration dependent manner and also, this effect was observed in LPS-preprimed cells. Concurrent incubation of cells with PKC inhibitors, H-7 or chelerythrine, and LPS resulted in the diminution of NO production regardless of glucose concentration but this was not in the case of LPS-prepriming, that is, chelerythrine showed a minimal effect on the glucose- enhancing effect. PMA, a PKC activator, did not show any significant effect on glucose-associated NO production. Modulation of sorbitol pathway with zopolrestat, an aldose reductase inhibitor, did not affect LPS-induced NO production and iNOS expression under high glucose condition. And also, sodium pyruvate, which is expected to normalize cytosolic $NADH/NAD^+$ ratio, did not show any significant effect at concentrations of up to 10 mM. Glucosamine marginally increased the endotoxin-induced nitrite release in both control and high glucose treated group. 6-diazo-5-oxonorleucine (L-DON) and azaserine, glutamine: fructose- 6-phosphate amidotransferase (GFAT) inhibitors, significantly diminished the augmentation effect of high glucose on endotoxin-induced NO production. On the other hand, negative modulation of GFAT inhibitors was not reversed by the treatment of glucosamine, suggesting the minimal involvement, if any, of glucosamine pathway in glucose-enhancing effect. In summary, these results strongly suggest that the hexosamine biosynthesis pathway and the activation of PKC via sorbitol pathway do not contribute to the augmenting effect of high glucose on endotoxin induced NO production in macrophage-like Raw264.7 cells.

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.2
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• pp.232-236
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• 2008

This experiment was performed to investigate the effects of two energy levels and four lysine:digestible energy (DE) ratios on the apparent digestibility of nutrients in finishing pigs. The experiment was conducted using a $2{\times}4$ randomized complete block (RCB) design with three replicates. Twenty-four cross-bred finishing barrows ((Landrace${\times}$Yorkshire)${\times}$Duroc) with an average body weight of $64.2{\pm}0.69kg$ were assigned to one of eight treatments. Each barrow was placed in an individual metabolism crate and dietary treatment and water was provided ad libitum. Diets were designed to contain lysine:ME ratios of 1.5, 1.8, 2.1 and 2.4 g/Mcal at 3.35 and 3.6 Mcal/kg of diet in a $4{\times}2$ factorial arrangement. Dry matter (DM), ash, Ca and P digestibility were not affected by energy density or lysine:DE ratios. Crude fat digestibility increased as the energy density increased from 3.35 to 3.6 Mcal of DE/kg. Increasing the lysine:DE ratio also increased crude protein digestibility. There were no interactions between energy density and lysine:DE ratio in terms of nutrient digestibility. Nitrogen excretion via feces was not affected by energy density and lysine:DE ratio, while nitrogen excretion via urine was significantly affected by energy density and lysine:DE ratio. The apparent digestibility of all amino acids except for isoluecine, arginine and aspartic acid as well as average values of essential amino (EAA), non-essential amino acids (NEAA) and total amino acid digestibility (p>0.05) were not affected by energy density. The apparent digestibility of all amino acids except for leucine, proline, alanine and tyrosine, NEAA and total amino acid digestibility were significantly affected by lysine: DE ratio (p<0.05). Interactive effects of energy and lysine:DE ratio also significantly affected amino acid digestibility except for isoleucine, alanine, cystine, leucine, phenylalanine, glutamine and proline (p<0.05). In conclusion, these results suggest that maintaining the appropriate lysine:DE ratio becomes more important as the energy density of the diet increases. Consequently, increasing the lysine:DE ratio can result in increased crude protein digestibility and urinary nitrogen excretion, although apparent protein digestibility and nitrogen excretion were not affected by energy density Furthermore, increasing the lysine:DE ratio also increased the apparent digestibility of essential amino acids, except for leucine, regardless of energy density. The optimum lysine:DE ratio for maximum essential amino acid digestibility of the $64.2{\pm}0.69kg$ pig is approximately 2.4 g of lysine/Mcal of DE.

• Food Science and Preservation
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• v.23 no.4
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• pp.547-552
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• 2016

The objective of this research was to find out the potential value of flaxseed as a dietary supplement as well as an edible oil resource. The characteristics of yellow flaxseed oil and brown flaxseed oil were compared to check which oil is better in the aspects of cooking purpose and of nutritional value. The quality of flaxseed oil was evaluated based on the composition of fatty acid, the content of phenolic compounds and the anti-oxidant activity. The total phenolics of yellow flaxseed oil and brown flaxseed oil were $10.78{\pm}0.46$ and $29.88{\pm}3.25mg/100g$, respectively. Their ${\gamma}$-tocopherol contents were 20.59 and 17.94 mg/100 g, respectively. Contents of linolenic acid were 56.60 and 31.38% and oleic acid were 18.24 and 39.16 %, respectively. Yellow flaxseed oil showed higher ratio of unsaturated fatty acid than brown flaxseed oil. However, brown flaxseed oil showed higher electron-donating abilities than brown flaxseed oil, which might be due to its higher content of phenolic compounds. In conclusion, flaxseed has a great potential as a good edible oil resource due to its high content of unsaturated fatty acid and anti-oxidant activity.

• Asian-Australasian Journal of Animal Sciences
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• v.13 no.5
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• pp.587-594
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• 2000

At present embryonic stem (ES) cells with confirmed pluripotential properties are only available in the mouse. Recently, we were able to isolate, culture and genetically transform primordial germ cell (PGC)-derived cells from pig embryos and demonstrate their ability to contribute to chimera development in the pig. In order to determine whether the system we developed could be used to isolate embryonic germ (EG) cells from other mammalian species, we placed isolated PGCs from cattle, goats, rabbits and rats in culture. Briefly, PGCs were isolated from fetuses of cow (day 30-50), goat (day 25), rabbit (day 15-18) and rat (day 11-12), and plated on STO feeder cells in Dulbecco's modified Eagle's medium (DMEM): Ham's F10 medium (1:1) supplemented with 0.01 mM nonessential amino acids, 2 mM L-glutamine, 0.1 mM $\beta$ - mercaptoethnol, soluble recombinant human stem cell factor (SCF; 40ng/ml), human basic fibroblast growth factor (bFGF; 20ng/ml) and human leukemia inhibitory factor (LIF; 20ng/ml). For maintenance of the cells, colonies were passed to fresh feeders every 7-10 days. In all species tested, we were able to obtain and maintain colonies with ES-like morphology. Their developmental potential was tested by alkaline phosphatase (AP) staining and in vitro differentiation assay. For genetic transformation, cells were electroporated with a construct containing the green fluorescent protein (GFP) under the control of the cytomegalovirus (CMV) promoter. GFP-expressing colonies were detected in cattle, rabbits and rats. These results suggest that PGC-derived cells from cattle, goats, rabbits and rats can be isolated, cultured, and genetically transformed, and provide the basis for analyzing their developmental potential and their possible use for the precise genetic modification of these species.

• Asian-Australasian Journal of Animal Sciences
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• v.18 no.7
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• pp.942-948
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• 2005

New polymorphism of major histocompatibility complex B-G genes was investigated by amplification and digestion of a 401bp fragment including intron 1 and exon 2 using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique with two restriction enzymes of Msp I and Tas I in eight breeds of Chinese indigenous chickens and one exotic breed. In the fragment region of the gene, three novel single nucleotide polymorphisms (SNPs) were detected at the two restriction sites. We found the transition of two nucleotides of A294G and T295C occurred at Tas I restriction site, and consequently led to a non-synonymous substitution of asparagine into serine at position 54 within the deduced amino acid sequence of immunoglobulin variable-region-like domain encoded by the exon 2 of B-G gene. It was observed at rare frequency that a single mutation of A294G occurring at the site, also caused an identical substitution of amino acid, asparagine 54-to-serine, to that we described previously. And the transversion of G319C at Msp I site led to a non-synonymous substitution, glutamine 62-to-histidine. The new alleles and allele frequencies identified by the PCR-RFLP method with the two enzymes were characterized, of which the allele A and B frequencies at Msp I and Tas I loci were given disequilibrium distribution either in the eight Chinese local breeds or in the exotic breed. By comparison, allele A at Msp I locus tended to be dominant, while, the allele B at Tas I locus tended to be dominant in all of the breeds analyzed. In Tibetan chickens, the preliminary association analysis revealed that no significant difference was observed between the different genotypes identified at the Msp I and Tas I loci and the laying performance traits, respectively.

• Korean Journal of Veterinary Research
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• v.39 no.4
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• pp.724-729
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• 1999

Amino acids, Na, K and glutathione (GSH) in red blood cells (RBCs) and hematological indices were examined in Korean dogs. A total of seven dogs possessing RBCs with high K and high glutathione (GSH)(HK/HG) were found in 42 Korean dogs : three from Cheju dog, two from Jindo dog and two from Korean mongrel. The RBCs in Korean HK/HG dog contained abnormally high aspartate (Asp), Glu and glutamine (Gln) the same as in HK/HG RBCs from Japanese Shiba dog. Two dogs possessing RBCs with HK and low GSH (HK/LG) were found in Cheju dog, and they accumulated Asp and Gln. Thus, not only the existence of HK dog was confirmed in Korean dogs, but HK/LG dog was also found. The Asp concentration in RBCs from seven of 33 LK dogs was more than $1000{\mu}mol/lc$, the same as in variant LK RBCs with defective Glu/Asp transport (LK/GAT), while it was less than $800{\mu}mol/lc$ in normal LK RBCs. Thus, there were variant dogs having RBCs with abnormally high amino acids accumulation among HK and LK Korean dogs.

• Korean Journal of Food Science and Technology
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• v.45 no.3
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• pp.350-355
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• 2013

The quality of fermented dropwort extract (FDE) and fermented dropwort vinegar (FDV) was assessed for free sugar, organic acid and free and total amino acid content. Major organic acids were lactic acid in FDE and acetic acid in FDV. Free sugars in FDE were fructose and glucose, and those in FDV were fructose, sucrose, and maltose. Aspartic acid was the major free amino acid in both FDE and FDV. Additionally, the main free amino acids in FDE were alanine and ${\gamma}$-amino-n-butyric acid (GABA), while those in FDV were arginine and valine. Moreover, to investigate the protective effects of FDE and FDV against oxidative stress induced by t-BHP and $H_2O_2$, C6 cells were treated with FDE or FDV prior to inducing the oxidative damage. FDE and FDV inhibited cell death significantly in a dose-dependent manner. These data imply that FDE and FDV may be effective in neuronal cell protection against oxidative damage.

• Journal of Life Science
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• v.16 no.5
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• pp.716-722
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• 2006

Superoxide dismutase (SOD) is physiologically important in regulating cellular homeostasis and apoptotic cell death, and its mutations are the cause of familial amyotrophic lateral sclerosis (FALS). Mitochondrial serine protease HtrA2 has a pro-apoptotic function and has known to be associated with neurodegenerative disorders. To investigate the relationship between genes associated with apoptotic cell death, such as HtrA2 and SOD1, we utilized the pGEX expression system to develop a simple and rapid method for purifying wild-type and ALS-associated mutant SOD1 proteins in a suitable form for biochemical studies. We purified SOD1 and SOD1 (G93A) proteins to approximately 90% purity with relatively high yields (3 mg per liter of culture). Consistent with the result in mammalian cells, SOD1 (G93A) was more insoluble than wild-type SOD1 in E. coli, indicating that research on the aggregate formation of SOD1 may be possible using this pGEX expression system in E. coli. We investigated the HtrA2 serine protease activity on SOD1 to assess the relationship between two proteins. Not only wild-type SOD1 but also ALS-associated mutant SOD1 (G93A) were cleaved by HtrA2, resulting in the production of the 19 kDa and 21 kDa fragments that were specific for anti-SOD1 antibody. Using protein gel electrophoresis and immunoblot assay, we compared the relative molecular masses of thrombin-cleaved GST-SOD1 and HtrA2-cleaved SOD1 fragments and can predict that the HtrA2-cleavage sites within SOD1 are the peptide bonds between leucine 9-lysine 10 (L9-K10) and glutamine 23-lysine 24 (Q23-K24). Our study indicates that SOD1 is one of the substrate for HtrA2, suggesting that both HtrA2 and SOD1 may be important for modulating the HtrA2-SOD1-mediated apopotic cell death that is associated with the pathogenesis of neurodegenerative disorder.

• Journal of Plant Biotechnology
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• v.38 no.2
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• pp.186-190
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• 2011

This study was conducted to evaluate effects of various kinds or concentrations in abscisic acid (ABA), reduced nitrogen sources (casein hydrolysate, casamino acid and L-glutamine) and osmoticum for production of somatic embryos (SEs) from pro-embryogenic mass (PEM) in yellow poplar (Liriodendron tulipifera). In comparison of various concentrations of ABA, the highest number (640/10 mg PEM) of SEs was marked in the treatment of 0.5 mg/L. With higher concentration than 0.5 mg/L ABA, number of induced SEs were decreased. And the lowest number of SEs were obtained from the treatment of 20 mg/L ABA. Differences of 8 treatments of the nitrogen sources in the medium were also compared. In the experiment of 8 treatments for SEs production, the highest result showed in the treatment of 500 mg/L casamino acid (223/5 mg PEM). In comparison of different kinds/concentrations of osmotica for SEs induction, the best response was obtained from the treatment of 4% sucrose (317/5 mg PEM). In contrast, no SEs were found from the treatments supplemented with any concentrations of maltose.

• Korean Journal of Food Science and Technology
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• v.22 no.1
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• pp.88-93
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• 1990

For effective separation of the N-TFA n-butyl ester amino acids on the stainless steel column by GLC, dual column of the mixed stationary phases, 3.36% OV-17+3.0% SE-30(column 1) and 1% NPGS +0.5% OV-17+0.5% SE-30(column 2) on chromosorb W HP 100-120 mesh, were used. On the column 1. the nineteen amino acids except histidine were obtained. However, alanine and valine peaks were not separated by this column. On the column 2, the sixteen amino acid peaks showed good separation, but tryptophan. arginine, histidine, and tyrosine peaks were not obtained. Calibration graphs for all amino acids obtained by the plotting the ratios of their peaks hights to that of internal standard versus the micro mole of the amino acids in the range $1.25{\times}10^{-3}{\mu}mol-1.0{\times}10^{-2}{\mu}mole$ showed linearity and passed through the origin.