• Asian-Australasian Journal of Animal Sciences
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• 제8권4호
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• pp.393-402
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• 1995

Experiments were conducted to evaluate the nutritive values of supplemental L-lysine, liquid and powder type, and DL-methionine in weanling pigs. For feeding trial, 165 weanling pigs were treated in 2 controls; 18 and 16% CP, 6 supplementations of lysine alone to 16% CP diets; 0.1, 0.2 and 0.4% of liquid and powder type each, and 3 supplementations of lysine + methionine to 15% CP diets; 0.05 + 0.025, 0.1 + 0.05 and 0.2 + 0.1%. Pigs were fed for 5 week to investigate the protein sparing effect of supplemental amino acid, and the optimal supplemental level. A metabolic trial included the measurements of digestibilities of dry matter, crude protein, crude fat, crude fiber, energy, phosphorus and amino acids. The liver acinar cell culture was conducted for the protein synthesis activity of the pigs fed each experimental diet. Supplementation of both type of L-lysine in 16% CP diet showed improved daily weight gain and feed efficiency which were compatible with those of pigs fed 18% CP diet. Groups fed liquid lysine did not differ from those fed powder type in growth performance. Supplementation of lysine and methionine to 15% CP diet did not improve growth performance of pigs to the extent that 18% CP diet was fed. In nutrient digestibility, 16% CP control diet showed significantly (p < 0.05) lower crude protein digestibility than any other treatments. Digestibilities of 16% CP diets with lysine supplementation were equal to that of 18% CP control, while digestibilities of 15% CP diets with the supplementation of lysine + methionine was inferior to that of 18% CP control. Supplementation of lysine alone reduced the nitrogen excretion compared to the none supplemented control groups. However, addition of lysine + methionine excreted more nitrogen than controls. Pigs fed diet supplemented with lysine alone, or lysine + methionine excreted less fecal phosphorus than those fed none supplemetation. Retained protein from liver tissue of pigs fed 18% diet was significantly (p < 0.05) greater than those fed 16% CP diet. A significant difference (p < 0.05) was observed in physical type of lysine. Feeding of powder type showed less secreted protein and greater retained protein in the culture of liver acinar cell. It is concluded that supplementation of lysine at the level of 0.1 to 0.2% can spare 2% of dietary protein and reduce nitrogen excretion by 19.3%. Also, no difference in nutritional values was observed between liquid and powder lysine in weanling pigs.

• Radiation Oncology Journal
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• 제16권4호
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• pp.377-388
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• 1998

목적 : 방사선조사에 의한 apoptosis에서 나타나는 각종 세포주기관련 유전자들의 발현 양상을 RNA와 단백 수준에서 분석하여 방사선조사에 의한 apoptosis에서의 세포주기 조절의 변화를 규명함으로서 방사선치료의 기전에 대한 분자생물학적 이해를 도모하고자 본 연구를 시행하였다. 대상 및 방법 : promyelocytic leukemia 세포주인 HL60 세포주를 배양하여 선형가속기(6MV X-선)로 세포에 8Gy의 방사선을 조사하였다. 조사후 다양한 시간 간격으로 Apoptotic DNA Fragmentation Assay법을 이용하여 apoptosis를 확인하고 동시에 세포주기관련 유전자들(cyclinA, cyclin B, cyclin C, cyclin Dl, cyclin E, cdc2, CDK2, CDK4, $p16^{INK4a}$, $p21^{WAF1}$, $p27^{KIP1}$, E2F, PCNA와 Rb)을 단백질과 RNA 수준에서 분석하기위해 western blot analysis와 반정량적 RT-PCR을 시행하였다. 결과: 8 Gy의 방사선 조사에 의해 HL60세포에서 apoptosis가 관찰 되었다. 방사선 조사군에서 cyclin A단백은 조사후 48시간까지 시간이 갈수록 증가하였으며, cyclin E, E2F, CDK2 및 Rb 단백은 증가되었다가 다시 감소를 보였다. Rb단백의 증가는 대부분 비활성형인 ppRb (phosphorylated Rb protein)의 양적변화에 의한 것이었다. cyclin Dl, PCNA, COC2, CDK4, $p16^{INK4a}$단백은 발현의 차이를 보이지 않았으며 $p21^{WAF1}$과 $p27^{KIP1}$ 단백은 검출되지 않았다. cyclin A, B, C mRNA는 방사선 조사 직후 감소하였다가 12시간부터 발현이 증가되었으며 cyclin Dl mRNA는 조사후 바로 증가하여 48시간에 다시 감소하였다. cyclin E mRNA는 조사 후 시간이 경과함에 따라 감소하였다. CDK2 mRNA는 3시간째는 감소하다가 6시간부터 많은 증가를 보였으며 CDK4 mRNA는 조사후 6-12시간에 급격한 발현증가를 보였다. $p16^{INK4a}$ RNA는 발현의 변화가 없었으며, $p21^{WAF1}$ 및 $p27^{KIP1}$ RNA의 발현은 관찰되지 않았다. 결론 : 이상의 결과로 미루어볼 때, 방사선 조사에 의한 HL60세포의 apoptosis와 세포의 Gl/S transition는 밀접한 관계가 있는 것으로 생각되며 Rb단백의 증가와 활성형 Rb단백의 감소 현상도 관련이 있는 것으로 사료된다. 이는 E2F의 비정상적인 과발현 및 cyclin E/CDK2의 발현 증가와 관련이 있는 것으로 추측된다. 또한 $p21^{WAF1}$ 및 $p27^{KIP1}$는 방사선에 의한 apoptosis에는 관여되지 않는 것으로 사료된다.

• Animal Bioscience
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• 제36권10호
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• pp.1584-1595
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• 2023

Objective: This study was conducted to evaluate the effect of a low-protein diet on growth performance, carcass traits, nutrient digestibility, blood profiles, and odor emissions in growing-finishing pigs. Methods: A total of 126 crossbred pigs ([Yorkshire×Landrace]×Duroc) with an average body weight (BW) of 38.56±0.53 kg were used for a 14-week feeding trial. Experimental pigs were allotted to one of 6 treatments in 3 replicates of 7 pigs per pen in a randomized complete block design. Pigs were fed each treatment diet with different levels of crude protein (CP). Phase 1 (early growing): 14%, 15%, 16%, 17%, 18%, 19%; phase 2 (late growing): 13%, 14%, 15%, 16%, 17%, 18%; phase 3 (early finishing): 12%, 13%, 14%, 15%, 16%, 17%; phase 4 (late finishing): 11%, 12%, 13%, 14%, 15%, 16%. All experimental diets in each phase were contained the same concentration of lysine (Lys), methionine (Met), threonine (Thr), and tryptophan (Trp). Results: Over the entire experimental period, there was no significant difference in BW, average daily feed intake, and gain-to-feed ratio among all treatments (p>0.05), but a quadratic effect (p = 0.04) was observed in average daily gain (ADG) during the late finishing phase with higher ADG in Group D. Blood urea nitrogen concentration linearly increased with an increase in dietary CP levels (p<0.01). Regarding nutrient digestibility, excreted nitrogen in urine and feces and nitrogen retention linearly increased as the CP level increased (p<0.01). A linear effect was observed with increasing CP levels in amines, ammonia, and hydrogen sulfide in odor emissions (p<0.01). No significant effects were observed in the measurements of carcass traits and meat characteristics (p>0.05). Conclusion: In phase feeding, reducing the CP level to 14% in early-growing pigs, 13% in late-growing pigs, 12% in early-finishing pigs, and 11% in late-finishing pigs is recommended.

• Asian Pacific Journal of Cancer Prevention
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• 제16권9호
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• pp.3759-3765
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• 2015

Background: Approaches in disruption of MDM2-p53 interactions have now emerged as an important therapeutic strategy in resurrecting wild type p53 functional status. The present study highlights virtual screening strategies in identification of high affinity small molecule non-peptidic inhibitors. Nutlin3A and RG7112 belonging to compound class of Cis-imidazoline, MI219 of Spiro-oxindole class and Benzodiazepine derived TDP 665759 served as query small molecules for similarity search with a threshold of 95%. The query molecules and the similar molecules corresponding to each query were docked at the transactivation binding cleft of MDM2 protein. Aided by MolDock algorithm, high affinity compound against MDM2 was retrieved. Patch Dock supervised Protein-Protein interactions were established between MDM2 and ligand (query and similar) bound and free states of p53. Compounds with PubCid 68870345, 77819398, 71132874, and 11952782 respectively structurally similar to Nutlin3A, RG7112, Mi219 and TDP 665759 demonstrated higher affinity to MDM2 in comparison to their parent compounds. Evident from the protein-protein interaction studies, all the similar compounds except for 77819398 (similar to RG 7112) showed appreciable inhibitory potential. Of particular relevance, compound 68870345 akin to Nutlin 3A had highest inhibitory potential that respectively showed 1.3, 1.2, 1.16 and 1.26 folds higher inhibitory potential than Nutilin 3A, MI 219, RG 7112 and TDP 1665759. Compound 68870345 was further mapped for structure based pharamacophoric features. In the study, we report Cis-imidazoline derivative compound; Pubcid: 68870345 to have highest inhibitory potential in blocking MDM2-p53 interactions hitherto discovered.

• Asian-Australasian Journal of Animal Sciences
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• 제14권4호
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• pp.507-514
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• 2001

Two experiments were carried out to investigate whether duckweed is useful as a dietary protein source for fine-wool Merino sheep and to evaluate its effects on wool yield and characteristics. In Experiment 1, the sheep were given one of three maintenance diets consisting of oaten chaff (520-700 g/d) supplemented with 16-32 g crude protein/d in the form of fresh (1 kg/d) or sun-dried (50-100 g/d) duckweed. Each ration was estimated to provide 5.4 MJ (1.3 Mcal)/d of metabolisable energy (ME). The sheep readily ingested the fresh or dried duckweed. None of the wool measures (yield, rate of fibre elongation, fibre diameter) differed (p>0.05) between dietary treatments. In Experiment 2, oaten-chaff-based diets (800 g/d) supplying 6.5-7.2 MJ (1.6-1.7 Mcal)/d of ME were supplemented with iso-nitrogenous amounts (4-5 g N) either of urea (8 g), cottonseed meal (60 g) or dried duckweed (100 g). In this experiment, the rate of wool fibre elongation, thought to be related to intestinal amino acid absorption, was lower (p<0.05) for sheep given the oaten chaff/urea diet than for those given either oaten chaff/cottonseed meal or oaten chaff/duckweed for which the rates did not differ (p>0.05). Fibre diameter, which ranged from 16.0-16.7 mm, did not differ (p>0.05) between diets, but tended to be lower on the oaten chaff/urea diet so that volume of wool produced was also significantly lower (p<0.05) on this diet than on the diets containing duckweed or cottonseed meal. Rumen ammonia concentrations at 4.5 and 7.5 h after feeding were higher (p<0.05) for sheep given the oaten chaff/urea diet than for those given the other two diets. A comparison of the rumen ammonia concentrations, wool growth rate and predicted flows of amino acids from the rumen of sheep supplemented with duckweed rather than cottonseed meal suggested that duckweed is a valuable source of 'escape protein' for ruminants.

• Asian-Australasian Journal of Animal Sciences
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• 제23권3호
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• pp.333-339
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• 2010

The objectives of this study were to use the NRC-2001 model and DVE/OEB system to model potential nutrient supply to ruminants and to compare frost damaged (also called "frozen" wheat with normal wheat. Quantitative predictions were made in terms of: i) Truly absorbed rumen synthesized microbial protein in the small intestine; ii) Truly absorbed rumen undegraded feed protein in the small intestine; iii) Endogenous protein in the digestive tract; iv). Total truly absorbed protein in the small intestine; and v). Protein degraded balance. The overall yield losses of the frozen wheat were 24%. Results showed that using the DVE/OEB system to predict the potential nutrient supply, the frozen wheat had similar truly absorbed rumen synthesized microbial protein (65 vs. 66 g/kg DM; p>0.05), tended to have lower truly absorbed rumen undegraded feed protein (39 vs. 53 g/kg DM; p<0.10) and had higher endogenous protein (14 vs. 9 g/kg DM; p<0.05). Total truly absorbed protein in the small intestine was significantly lower (89 vs. 110 g/kg DM, p<0.05) in the frozen wheat. The protein degraded balance was similar and both were negative (-2 vs. -1 g/kg DM). Using the NRC-2001 model to predict the potential nutrient supply, the frozen wheat also had similar truly absorbed rumen synthesized microbial protein (average 56 g/kg DM; p>0.05), tended to have lower truly absorbed rumen undegraded feed protein (35 vs. 48, g/kg DM; p<0.10) and had similar endogenous protein (average 4 g/kg DM; p>0.05). Total truly absorbed protein in the small intestine was significantly lower (95 vs. 108 g/kg DM, p<0.05) in the frozen wheat. The protein degraded balance was not significantly different and both were negative (-16 vs. -19 g/kg DM). In conclusion, both models predict lower protein value and negative protein degraded balance in the frozen wheat. The frost damage to the wheat reduced nutrient content and availability and thus reduced nutrient supply to ruminants by around 12 to 19%.

• Asian-Australasian Journal of Animal Sciences
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• 제27권6호
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• pp.797-805
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• 2014

The effect of concentrate mixtures with crude protein (CP) levels 10%, 13%, 16%, and 19% and diets with roughage to concentrate ratios 80:20, 60:40, 40:60, and 20:80 (w/w) were determined on dry matter (DM) and organic matter (OM) digestibility, and fermentation metabolites using an in vitro fermentation technique. In vitro fermented attributes were measured after 4, 24, and 48 h of incubation respectively. The digestibility of DM and OM, and total volatile fatty acid (VFA) increased whereas pH decreased with the increased amount of concentrate in the diet (p<0.001), however CP levels of concentrate did not have any influence on these attributes. Gas production reduced with increased CP levels, while it increased with increasing concentrate levels. Ammonia nitrogen ($NH_3$-N) concentration and microbial CP production increased significantly (p<0.05) by increasing CP levels and with increasing concentrate levels in diet as well, however, no significant difference was found between 16% and 19% CP levels. Therefore, 16% CP in concentrate and increasing proportion of concentrate up to 80% in diet all had improved digestibility of DM and organic matter, and higher microbial protein production, with improved fermentation characteristics.

• 한국가축번식학회지
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• 제26권1호
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• pp.73-78
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• 2002

젖소 난소 황체에 있어서 중심강의 유무에 따른 황체 조직 중의 total protein, DNA 및 RNA 함량을 조사하여 중심강이 있는 황체와 중심강이 없는 황체간의 기능성을 구명하고자 수행한 시험에서 다음과 같은 결과를 얻었다. 1. 중심강이 없는 황체의 total, supernatant 및 Pellt protein의 함량은 각각 32.83, 16.87 및 15.96 mg/g wet tissue이었고, 중심강이 있는 황체의 그것들은 각각 29.62, 16.10 및 13.52 mg/g wet tissue으로서 중심강이 없는 황체와 중심강이 있는 황체간에 유의적인 차이를 나타내지 않았다(p〉0.05). 2. DNA 함량은 중심강이 없는 황체가 1.99mg/g wet tissue이었고 중심 강이 있는 황체가 1.32mg/g wet tissue으로 중심강이 없는 황체와 있는 황체간에 유의적인 차이(p＜0.05)를 나타내었다. Protein : DNA ratios에 있어서도 중심강이 없는 황체가 16.63mg/g wet tissue이었고 중심강이 있는 황체가 22.99mg/g wet tissue으로 중심강이 없는 황체와 중심강이 있는 황체간에 유의적인 차이(p＜0.05)를 나타내었다. 3. 중심강이 없는 황체의 RNA함량, Protein: RNA 및 RNA:DNA ratios는 각각 2.87, 12.24및 1.43mg/g wet tissue이었고, 중심강이 있는 황체의 그것들은 각각 2.47, 13.73 및 1.89mg/g wet tissue으로서 중심강이 없는 황체와 중심강이 있는 황체간에 유의적인 차이를 나타내지 않았다(p〉0.05).

• Bulletin of the Korean Chemical Society
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• 제16권5호
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• pp.401-406
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• 1995

The P. fragi lipase overexpressed in E. coli as a fusion protein of 57 kilodalton (kDa) has been purified through glutathione-agarose affinity chromatography by elution with free glutathione. The general properties of the purified GST-fusion protein were characterized by observing absorbance of released p-nitrophenoxide at 400 nm which was hydrolyzed from the substrate p-nitrophenyl palmitate. The optimum condition was observed at 25 $^{\circ}C$, pH 7.8 with 0.4 ${\mu}g$ of protein and 1.0 mM substrate in 0.6% (v/v) TritonX-100 solution. Also the lipase was activated by Ca+2, Mg+2, Ba+2 and Na+ but it was inhibited by Co+2 and Ni+2. pGEX-2T containing P. fragi lipase gene as expression vector was named pGL191 and used as a template for the site-directed mutagenesis by sequential PCR steps. A Ser-His-Asp catalytic triad similar to that present in serine proteases may be present in Pseudomonas lipase. Therefore, the PCR fragments replacing Asp217 to Arg and His260 to Arg were synthesized, and substituted for original fragment in pGL19. The ligated products were transformed into E. coli NM522, and pGEX-2T harboring mutant lipase genes were screened through digestion with XbaI and StuI sites created by mutagenic primers, respectively. No activity of mutant lipases was observed on the plate containing tributyrin. The purified mutant lipases were not activated on the substrate and affected at pH variation. These results demonstrate that Asp217 and His260 are involved in the catalytic site of Pseudomonas lipase.

• 한국식품과학회지
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• 제24권3호
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• pp.284-288
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• 1992

영양 저해인자로 알려진 phytate를 제거한 분리 유채단백(low-phytate rapeseed protein isolate)을 제조하여 유채단백질과 phytate와의 결합양상에 미치는 pH 및 phytate 첨가량의 영향을 알아보기 위하여 SDS-polyacrylamide gel 전기영동법을 수행하였다. 가용화된 분리 유채단백은 pH2.0과 pH5.0에서 phytate 첨가량이 증가할수록 가용성 단백질 band의 수가 감소하였으며 pH11.5에서는 band의 변화가 없었다. pH2.0에서 대조군의 18개 band중 7개의 band(105.8, 52.3, 37.3, 34.8, 26.3, 21.3, 18.4KDa)가 100 mg phytate 첨가시 침전되었으며 150 mg 첨가시에는 6개의 band(78.8, 46.5, 19.4, 16.8, 11.7, 8.5KDa)가 더 사라졌다. pH5.0에서는 대조군과 비교하여 15개의 band중 4개의 band(34.8, 21.3, 18.4, 16.8KDa)만이 phytate 첨가시 침전되었다. 따라서 유채단백을 식품소재로 이용하기 위해서는 phytate 함량을 낮춤으로써 그의 기능성을 향상시킬 수 있을 것이다.