• Asian-Australasian Journal of Animal Sciences
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• v.23 no.3
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• pp.292-301
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• 2010

Follicle-stimulating hormone (FSH) is a pituitary glycoprotein hormone that is encoded by separate alpha- and betasubunit genes. It plays a key role in stimulating and regulating ovarian follicular development and egg production in chicken. FSH signal transduction is mediated by the FSH receptor (FSHR) that exclusively interacts with the beta-subunit of FSH, but characterization of prokaryotic expression of the FSHb gene and its effect on the expression of the FSHR gene in local chickens have received very little attention. In the current study, the cDNA fragment of the FSHb gene from Dagu chicken was amplified using reverse transcription polymerase chain reaction (RT-PCR), and inserted into the pET-28a (+) vector to construct the pET-28a-FSHb plasmid. After expression of the plasmid in E. coli BL21 (DE3) under inducing conditions, the recombination protein, $FSH{\beta}$ subunit, was purified and injected into the experimental hens and the effect on the mRNA expression levels of the FSHR gene was investigated. Sequence comparison showed that the coding region of the FSHb gene in the local chicken shared 99%-100% homology to published nucleotides in chickens; only one synonymous nucleotide substitution was detected in the region. The encoded amino acids were completely identical with the reported sequence, which confirmed that the sequences of the chicken FSHb gene and the peptides of the $FSH{\beta}$ subunit are highly conserved. This may be due to the critical role of the normal function of the FSHb gene in hormonal specificity and regulation of reproduction. The results of gene expression revealed that a recombinant protein with a molecular weight of about 19 kDa was efficiently expressed and it was identified by Western blotting analysis. After administration of the purified $FSH{\beta}$ protein, significantly higher expression levels were demonstrated in uterus, ovary and oviduct samples (p<0.05). These observations suggested that the expressed $FSH{\beta}$ protein possesses biological activity, and has a potential role in regulation of reproductive physiology in chickens.

• Asian-Australasian Journal of Animal Sciences
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• v.22 no.9
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• pp.1267-1278
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• 2009

Three beef steers fitted with permanent cannulae in the rumen and duodenum were used to determine the effects of protein supply from soyhulls (SH) and wheat bran (WB) on ruminal metabolism, blood metabolites, nitrogen metabolism, nutrient digestion and concentrations of soluble non-ammonia nitrogen (SNAN) in ruminal (RD) and omasal digesta (OD). In a 3${\times}$3 Latin square design, steers were offered rice straw and concentrates formulated either without (control) or with two brans to increase crude protein (CP) level (9 vs. 11% dietary DM for control and bran-based diets, respectively). The brans used were SH and WB that had similar CP contents but different ruminal CP degradability (52 vs. 80% CP for SH and WB, respectively) for evaluating the effects of protein degradability. Ruminal ammonia concentrations were higher for bran diets (p<0.01) than for the control, and for WB (p<0.001) compared to the SH diet. Similarly, microbial nitrogen and blood urea nitrogen were significantly increased (p<0.05) by bran and WB diets, respectively. Retained nitrogen tended (p<0.082) to be increased by SH compared with the WB diet. Intestinal and total tract CP digestion was enhanced by bran diets. In addition, bran diets tended (p<0.085) to increase intestinal starch digestion. Concentrations of SNAN fractions in RD and OD were higher (p<0.05) for bran diets than for the control, and for WB than for the SH diet. More rumendegraded protein supply resulting from a higher level and degradability of CP released from SH and WB enhanced ruminal microbial nitrogen synthesis and ruminal protein degradation. Thus, free amino acids, peptides and soluble proteins from microbial cells as well as degraded dietary protein may have contributed to increased SNAN concentrations in the rumen and, consequently, the omasum. These results indicate that protein supply from SH and WB, having a low level of protein (13 and 16%, respectively), could affect ruminal metabolism and nutrient digestion if inclusion level is relatively high (>20%).

• Asian-Australasian Journal of Animal Sciences
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• v.24 no.5
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• pp.670-677
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• 2011

The growth rate of the young pig is generally much less than it's potential and may be constrained by endocrine status as well as nutrient intake. Growth factors are present in relatively high quantities in colostrum and play an important part in gut development. It is possible that supplementation of colostrum protein isolate may stimulate gut and whole body growth in the pig. Eight male and 8 female (Large Whitex${\times}$Landrace) piglets were weaned at 1 d of age after each pig had obtained colostrum from their dam, and were trained to consume one of two liquid diets. The two diets were based on either a colostrum protein isolate (n = 4 males and 4 females) or whey protein concentrate (n = 4 males and 4 females) and were formulated to contain equal levels of crude protein and amino acids. Pigs were fed their diets ad libitum for 28 days after which time 12 pigs were euthanised and various tissues and organs weighed. Pigs were bled for IGF-I analyses at 21 and 28 days of age. Daily gain was higher in pigs consuming the colostrum isolate (171 vs. 216 g/d, p = 0.010), particularly between 2 and 4 weeks of age (212 vs. 298 g/d, p = 0.010). Pigs tended to consume more of the liquid feed containing colostrum isolate (25.5 vs. 29.1 kg, p = 0.074) and gained more live weight per unit of liquid feed (0.203 vs. 0.223 g/g, p = 0.056). There were no effects of sex on growth performance. Pigs consuming the diet supplemented with colostrum isolate had higher (p<0.05) full gut weight (445 vs. 554 g, p = 0.026), empty gut weight (356 vs. 463 g, p = 0.008), stomach weight (42.2 vs. 54.4 g, p = 0.001), small intestine weight (222 vs. 275 g, p = 0.025) and large intestine weight (63.7 vs. 98.0 g, p = 0.005). Plasma IGF-I (99 vs. 150 ng/ml, p<0.001) and IGF-II (265 vs. 406 ng/ml, p<0.001) were higher in pigs fed colostrum isolate. Pigs consuming colostrum protein isolate ate more, grew faster and had higher plasma IGF-I concentrations than pigs consuming a diet with similar macronutrient content but devoid of growth factors.

• Journal of the Korean Chemical Society
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• v.42 no.3
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• pp.306-314
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• 1998

Acetolactate synthase (ALS) is the common enzyme in the biosynthesis of branched chain amino acids, Val, Leu, and Ile in bacteria, yeast, and higher plants. The enzyme is target site of several classes of structually diverse herbicides, including the sulfonylureas, the imidazolinones, the triazolopyrimidines, and the primidyl-oxy-benzoates. We have synthesized new triazolopyrimidine (TP) derivatives, and determined their inhibitory activities on barley ALS. $lC_{50}$ values for the active compounds were 3.2 nM-0.62 mM, and some of them appeared to be potent inhibitors. The progress curves for inhibition of ALS by TP4, a representative derivative, indicated that the extent of inhibition increased with incubation time. The inhibition of ALS by TP4 showed mixed-type inhibition with respect to pyruvate. Dual inhibition analyses of TP4 versus imidazolinone Cadre and feedback inhibitor Leu suggested that three different classes of inhibitors bind to ALS in a mutually exclusive manner. Chemical modification of tyrosyl residues of ALS decreased sensitivity of ALS to TP4, while modification of tryptophan and cysteine did not affect the sensitivity.

• Journal of the Korean Chemical Society
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• v.40 no.8
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• pp.542-548
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• 1996

Electrodes modified with threonine, methionine and serine as ligands, which are incorporated by ion exchange into a polycationic film of electropolymerized $[Ru(v-bpy)_3]^{2+}$, have been employed in the determination of mercury in solution. The redox response of the surface-immobilized mercury/ligand complex was used as the analytical signal. When the polymeric film was electropolymerized, the supporting electrolytes were TBAP and $KPF_6$ to compare the morphology and anodic stripping of resulted polymer electrodes. At the case of the latter, the film had high porosity to give an easy incorporation of dopant anions into polymeric film matrix and a high sensitivity in determination of mercury ion. Especially, this polymer modified electrode exhibited possibility of multiple use in mercury determination over ten times. In all cases, calibration curves which were plotted by log of the surface coverage-normalized redox response vs. log[Hg] exhibited an excellent correlation (r=0.99) for mercury concentrations ranging from 1.0{\times}10^{-8}{\sim}1.0{\times}10^{-2}M$. At these curves relative standard deviation was 5∼8% and saturation response was not observed at high concentration region. Serine of the employed ligands had the best sensitivity in analytical application, which had greater stability constant in forming a complex with mercury than others as $pK_{Hg}=8.54$. The formation constants of threonine and methionine were respectively 7.04 and 7.80.

• Korean Journal of Agricultural Science
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• v.24 no.1
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• pp.29-40
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• 1997

Six different types of teas from box thorn leaves collected on mid May were prepared by using different tea making procedures. The selected physicochemical propentes and the sensory qualities of the 6 different teas were analyzed. The box thorn leaves contained 87.71% moisture, 0.69% crude fat, 4.77% crude protein, 2.13% fibers, 3.11% carbohydrates and 1.59% ash. Based on the sensory analysis, roasted tea(duckum tea) had the highest values in taste, flavor, color and overall preference. Eighteen free amino acids were identified in the roasted tea, showing the specially high levels of proline(170.15mg%) and glutamic acid(112.34mg%). The roasted tea also contained glucose(1.07%), maltose(0.87%), sucrose(0.63%) and fructose(0.55%). The content of citric acid(337.43mg%) in the roasted tea was the highest, followed by malonic acid(54.17mg%), oxalic acid(48.66mg%), malic acid(27.41mg%), succinic acid(4.48mg%), fumaric acid(1.08mg%) and lactic acid(trace amount), in a decreasing order.

• Journal of Ginseng Research
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• v.44 no.3
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• pp.461-474
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• 2020

Background: Ginseng effectively reduces fatigue in both animal models and clinical trials. However, the mechanism of action is not completely understood, and its molecular targets remain largely unknown. Methods: By screening for proteins that interact with the primary components of ginseng (ginsenosides) in an affinity chromatography assay, we have identified muscle-type creatine kinase (CK-MM) as a potential target in skeletal muscle tissues. Results: Biolayer interferometry analysis showed that ginsenoside metabolites, instead of parent ginsenosides, had direct interaction with recombinant human CK-MM. Subsequently, 20(S)-protopanaxadiol (PPD), which is a ginsenoside metabolite and displayed the strongest interaction with CK-MM in the study, was selected as a representative to confirm direct binding and its biological importance. Biolayer interferometry kinetics analysis and isothermal titration calorimetry assay demonstrated that PPD specifically bound to human CK-MM. Moreover, the mutation of key amino acids predicted by molecular docking decreased the affinity between PPD and CK-MM. The direct binding activated CK-MM activity in vitro and in vivo, which increased the levels of tissue phosphocreatine and strengthened the function of the creatine kinase/phosphocreatine system in skeletal muscle, thus buffering cellular ATP, delaying exercise-induced lactate accumulation, and improving exercise performance in mice. Conclusion: Our results suggest a cellular target and an initiating molecular event by which ginseng reduces fatigue. All these findings indicate PPD as a small molecular activator of CK-MM, which can help in further developing better CK-MM activators based on the dammarane-type triterpenoid structure.

• Journal of Life Science
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• v.28 no.2
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• pp.162-169
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• 2018

Many cytoplasmic proteins are targeted to the cytoplasmic membrane of the trans-Golgi network (TGN) via an N-terminal short helix. We previously showed that the 20 N-terminal amino acids of Aplysia phosphodiesterase 4 (ApPDE4) long form are sufficient for its targeting to the plasma membrane and the TGN. The N-terminus of the ApPDE4 long form binds to PI4P and sulfatide in vitro. Therefore, in order to decipher the roles of sulfatide in Golgi complex targeting, we examined the cellular localization of sulfatide-binding peptides. In this study, we found that enhanced green fluorescent protein (EGFP) fused to the C-terminus of modified sulfatide- and heparin-binding peptides (mHSBP-EGFP) was localized to the TGN. On the other hand, its mutant, in which tryptophan was replaced with an alanine, leading to the impairment of heparin and sulfatide binding, was localized to cytosol. We also found that the TGN targeting of mHSBP-EGFP is impaired by the treatment of antimycin A, phenylarsine oxide (PAO), and adenosine but not a high concentration of wortmannin. These results suggest that PAO and adenosine-sensitive kinases, including phosphatidylinositol 4-kinase II, may play key roles in the recruitment of mHSBP-EGFP.

• Journal of Plant Biotechnology
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• v.44 no.1
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• pp.69-75
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• 2017

Dormancy-associated protein (DRM) is involved in the dormancy physiology of plants and is conserved in almost all plant species. Recent studies found that DRM genes are involved in the abiotic stress response, and characterization studies of these genes have been conducted in several plants. However, few studies have focused on DRM genes in woody plants. Therefore, in this study, cDNA coding for DRM (PagDRM1) was isolated from poplar (Populus alba ${\times}$ P. glandulosa), and its structure and expression characteristics were investigated. PagDRM1 encodes a putative protein composed of 123 amino acids, and the protein contains two conserved domains (Domain I and Domain II). PagDRM1 is present as one or two copies in the poplar genome. Its expression level was highest in the stem, followed by mature leaves, roots, and flowers. During the growth of cultured cells in suspension, PagDRM1 was highly expressed from the late-exponential phase to the stationary phase. In addition, PagDRM1 expression increased in response to drought, salt stress, and treatment with plant hormones (e.g., abscisic acid and gibberellic acid). Therefore, we suggested that PagDRM1 not only plays an important role in the induction of dormancy, but also contributes to stress tolerance in plants.

• Journal of Plant Biotechnology
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• v.44 no.1
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• pp.61-68
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• 2017

Gibberellic Acid-Stimulated Arabidopsis (GASA) genes are involved in plant hormone signaling, cell division and elongation, as well as in responses to stress conditions in plants. In this study, we isolated a GASA gene from hybrid poplar (Populus alba ${\times}$ P. glandulosa) and analyzed its physiological phenotype and molecular functions in poplar. PagGASA cDNA encodes a putative protein composed of 95 amino acids containing an N-terminal signal peptide and a conservative cysteine-rich C-terminal domain. Southern blot analysis revealed that one or two copies of the PagGASA are present in the poplar genome. The PagGASA transcripts were highly detected in flowers and roots. Moreover, the expression of PagGASA was induced by growth hormone (gibberellic acid) and stress hormones (abscisic acid, jasmonic acid, and salicylic acid). By using transgenic analysis, we showed that the upregulation of PagGASA in poplar provides high tolerance to drought stress. Therefore, our results suggest that PagGASA plays an important role in drought stress tolerance via stress-related plant hormone signaling in poplar.