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Isolation and Characterization of mas1+ of Schizosaccharomyces pombe, a Homologue of Human CIP29/Hcc-1 Involved in the Regulation of Cell Division (세포분열에 관여하는 인간의 CIP29/Hcc1 유전자와 상동성을 가지는 분열형 효모의 새로운 유전자 mas1+의 특성분석)

  • Cha, Jae-Young;Shin, Sang-Min;Ha, Se-Eun;Lee, Jung-Sup;Park, Jong-Kun
    • Journal of Life Science
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    • v.21 no.12
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    • pp.1666-1677
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    • 2011
  • The regulation of gene expression plays an important role in cell cycle controls. In this study, a novel gene, the $mas1^+$($\underline{mi}$tosis $\underline{as}$sociated protein) gene, a homolog of human CIP29/Hcc1, was isolated and characterized from fission yeast Schizosaccharomyces pombe (S. pombe) using a gene-specific polymerase chain reaction. The isolated gene contained a complete open reading frame capable of encoding 245 amino acid residues with a typical promoter, as judged by nucleotide sequence analysis. It was also found that a PCB ($\underline{p}$ombe cell $\underline{c}$ycle $\underline{b}$ox) is located in the promoter region, which controls M-$G_1$ specific transcription in S. pombe. The quantitative analysis of the $mas1^+$ transcript against $adh1^+$ showed that the pattern of expression is similar to that of the septation index. Cytokinesis of mas1 mutant was greatly delayed at $25^{\circ}C$ and $36^{\circ}C$, and a large number of multi-septate cells were produced. The mas1 mutant had 2C, 4C and 6C DNA contents, as determined by FACS analysis. In addition, the number of multi-septate cells significantly increased. When cells were cultured in nitrogen starvation medium to increase proliferation, the abnormal phenotypes of mas1 mutant dramatically increased. These phenotypes could be rescued by an overexpression of the $mas1^+$ gene. The mas1 protein localized in the nuclei of S. pombe and human HeLa cells, as evidenced by Mas1-EGFP signals. The abnormal growth pattern and the morphology of mas1 mutant were complemented by a plasmid carrying human CIP29/Hcc-1cDNA. In addition, CIP29 /Hcc-1 transcript level increased in active cell proliferation stages in the developing mouse embryos. These results indicate that the $mas1^+$ ishomologous to the human CIP29/Hcc1 gene and is involved in cytokinesis and cell shape control.

Role of PKR and EGR-1 in Induction of Interleukin-S by Type B Trichothecene Mycotoxin Deoxynivalenol in the Human Intestinal Epithelial Cells (B형 트리코테센 곰팡이 독소 데옥시니발레놀에 의한 인체 장관 상피세포 염증성 인터루킨 8유도에서의 PKR과 EGR-1의 상호 역할 규명)

  • Park, Seong-Hwan;Yang, Hyun;Choi, Hye-Jin;Park, Yeong-Min;Ahn, Soon-Cheol;Kim, Kwan-Hoi;Lee, Soo-Hyung;Ahn, Jung-Hoon;Chung, Duk-Hwa;Moon, Yu-Seok
    • Journal of Life Science
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    • v.19 no.7
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    • pp.949-955
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    • 2009
  • Mucosal epithelia sense external stress signals and transmit them to the intracellular cascade responses. Ribotoxic stress-producing chemicals such as deoxynivalenol (DON) or other trichothecene mycotoxins have been linked with gastrointestinal inflammatory diseases by Fusarium-contamination. The purpose of this study was to test the hypothesis that DON evokes the epithelial sentinel signals of RNA-dependent protein kinase (PKR) and early growth response gene 1 (EGR-1), which together contribute to the pro-inflammatory cytokine interleukin 8 (IL-8) in human intestinal epithelial cells. PKR suppression by the dominant negative PKR expression attenuated DON-stimulated interleukin-8 production. Moreover, 1L-8 transcriptional activation by DON was also reduced by PKR inhibition in the human intestinal epithelial cells. Treatment with the PKR inhibitor also suppressed EGR-1 promoter activity, mRNA and protein induction, although mitogen-activated protein (MAP) kinases such as extracellular signal-regulated protein kinases (ERK) 1/2, p38, c-Jun N-terminal Kinase (INK) were little affected or even enhanced in presence of a PKR inhibitor. These patterns were also compared in the EGR-1-suppressed cells, which showed much more suppressed production of 1L-8. All things taken into consideration, DON-activated sentinel signals of EGR-1 via PKR mediated interleukin-8 production in human intestinal epithelial cells, which provide insight into the possible general mechanism associated with mucosal inflammation as an intestinal toxic insult by ribotoxic trichothecene mycotoxins.

Comparative Analysis of Anti-oxidative, Anti-inflammatory, Anti-allergy, and Whitening Effects of Different Solvent Extracts from Zizania latifolia (고장초 추출 용매의 에탄올 함량에 따른 항산화, 항염증, 항알러지, 미백 활성 비교 분석)

  • Park, Se-Ho;Lee, Jae-Yeul;Yang, Seun-Ah
    • Journal of Life Science
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    • v.27 no.9
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    • pp.994-1002
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    • 2017
  • This study was performed to evaluate the anti-oxidative, anti-inflammatory, anti-allergy, and whitening effects of Zizania latifolia ethanol extracts prepared from 5 different ethanol concentrations (10, 30, 50, 70, and 90%). As the ethanol concentration in the extraction solvent was increased, the radical scavenging activities also increased. The inhibitory activity of Z. latifolia ethanol extracts on nitric oxide (NO) production in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells tended to increase as the content of ethanol increased. The highest inhibitory activity was obtained with 70% ethanol extract. The antiallergy effects of Z. latifolia ethanol extracts were tested by measuring the release of ${\beta}-hexosaminidase$ in IgE-sensitized RBL-2H3 cells. The suppressive effect of Z. latifolia ethanol extracts increased in a dose-dependent manner as the proportion of ethanol increased, except for the 10% ethanol extract. Furthermore, the inhibitory effects of Z. latifolia ethanol extracts against melanin production in ${\alpha}-melanocyte$ stimulated hormone (MSH)-stimulated B16F0 cells increased as the ethanol ratio increased, and 70 and 90% ethanol extracts showed similar inhibitory activities to arbutin, a positive control, at $250{\mu}m$. The present study confirmed the efficacy of Z. latifolia ethanol extracts in various areas, demonstrating antioxidative, anti-inflammation, antiallergy, skin protective, and skin whitening effects, with no cytotoxicity. It could be used as a raw material in functional foods, as well as in cosmetics.

Detection of Lamivudine-Resistant Mutations of HBV DNA Polymerase Gene Using PCR-Direct Sequencing

  • Lee, Kyung-Ok;Lee, Hye-Jung;Byun, Ji-Young;Lee, Sung-Yeun;Kim, Jeong-Sook;Jung, Na-Young;Chung, Soo-Jin;Seong, Hye-Soon;Kim, Kyung-Tae
    • Korean Journal of Clinical Laboratory Science
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    • v.38 no.3
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    • pp.196-202
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    • 2006
  • Treatment of hepatitis B virus (HBV) with lamivudine is effective in suppressing virus replication and results in reduced inflammatory activity. However the most troublesome problem of lamivudine treatment is the emergence of lamivudine-resistant strains with amino acid substitution in the YMDD motif of DNA polymerase gene during the treatment. The aim of this study was to determine the mutation of YMDD motif (codon 552) and codon 528 in chronic HBV patients with lamivudine therapy using PCR-direct sequencing and to investigate the relationship between lamivudine mediated HBV mutation and HBeAg. HBV DNA was extracted from serum samples of HBV patients and amplified by nested PCR with two sets of primer pairs selected in HBV DNA polymerase gene. Amplified PCR product was analyzed by 2% agarose gel electrophoresis and direct sequencing. HBV mutation was detected in 124 out of 207 samples (60%). Single mutation was 50.8% for M552I, 43.5% for M552V, 5.7% for M552I/V and the L528M mutation was 67.0%. Double mutation was 43.6% for M552V/L528M, 33.1% for M552I/L528(wild type), 17.7% for M552I/L528M and 5.6% for M552I/V/L528M. Serine mutation at YMDD motif (M552S) was not found and the L528M mutation frequently accompanied M552V type. In this study, the typical difference of frequencies for HBV mutation depending on HBeAg was not found. Moreover, the PCR-direct sequencing method used in this study might be a powerful tool for the mutation study in clinical reference laboratories with high volume.

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Effects of the Fermentation Periods on the Qualities and Functionality of the Vegetable Fermentation Broths (식물자원 발효액의 품질과 기능성에 미치는 발효기간의 영향)

  • Kim, Na-Mi;Lee, Jong-Won;Do, Jae-Ho;Park, Chae-Kyu;Yang, Jae-Won
    • Korean Journal of Medicinal Crop Science
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    • v.13 no.6
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    • pp.293-299
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    • 2005
  • To determine optimal fermentation period of vegetables mixed with black sugar without innoculation of microorganisms, changes in chemical components, quality characteristics of the fermented broth and physiological functionality during the fermentation period were investigated, pH and $^{\circ}BX$ of the fermented broths were decreased gradually during the fermentation period. Except for radish, L and a color values of fermented broths were increased but b values were decreased during fermentation period. Viscosity of fermented broths of vegetables were decreased after 3 months of fermentation. ${\alpha}-Amylase$ activity in fermentation broth of brocolli, eggplant, cabbage, chicory, aralia, radish were increased to 460, 430, 180, 420, 560, 260 after six months fermentation period. In radish and tomato fermentation broth, invertase activity were increased to 200 and 460 units and cellulase activity were increased to 280 and 140 after six months fermentation period. The content of total phenolic compounds and electron donating ability were the highest after 2 to 4 months fermentation period and decreased thereafter. No significant level of tyrosinase inhibitory activity and SOD-like activity were observed. In the sensory evaluation test of aralia fermentation broth of droop, sweet and sour flavor and bitter, astringent taste were decreased during the fermentation period and droop tastes were highest in 3 months. In radish fermentation broth, radish flavor and pungent taste were decreased and sweet taste was increased during fermentation period. Acceptability in overall was the greatest after three months. Based on the results stated above, optimal fermentation period was appropriated 3 or 4 months.

The Limitation of Air Carriers' Cargo and Baggage Liability in International Aviation Law: With Reference to the U.S. Courts' Decisions (국제항공법상 화물.수하물에 대한 운송인의 책임상한제도 - 미국의 판례 분석을 중심으로 -)

  • Moon, Joon-Jo
    • The Korean Journal of Air & Space Law and Policy
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    • v.22 no.2
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    • pp.109-133
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    • 2007
  • The legal labyrinth through which we have just walked is one in which even a highly proficient lawyer could easily become lost. Warsaw Convention's original objective of uniformity of private international aviation liability law has been eroded as the world community ha attempted again to address perceived problems. Efforts to create simplicity and certainty of recovery actually may have created less of both. In any particular case, the issue of which international convention, intercarrier agreement or national law to apply will likely be inconsistent with other decisions. The law has evolved faster for some nations, and slower for others. Under the Warsaw Convention of 1929, strict liability is imposed on the air carrier for damage, loss, or destruction of cargo, luggage, or goods sustained either: (1) during carriage in air, which is comprised of the period during which cargo is 'in charge of the carrier (a) within an aerodrome, (b) on board the aircraft, or (c) in any place if the aircraft lands outside an aerodrome; or (2) as a result of delay. By 2007, 151 nations had ratified the original Warsaw Convention, 136 nations had ratified the Hague Protocol, 84 had ratified the Guadalajara Protocol, and 53 nations had ratified Montreal Protocol No.4, all of which have entered into force. In November 2003, the Montreal Convention of 1999 entered into force. Several airlines have embraced the Montreal Agreement or the IATA Intercarrier Agreements. Only seven nations had ratified the moribund Guatemala City Protocol. Meanwhile, the highly influential U.S. Second Circuit has rendered an opinion that no treaty on the subject was in force at all unless both affected nations had ratified the identical convention, leaving some cases to fall between the cracks into the arena of common law. Moreover, in the United States, a surface transportation movement prior or subsequent to the air movement may, depending upon the facts, be subject to Warsaw, or to common law. At present, International private air law regime can be described as a "situation of utter chaos" in which "even legal advisers and judges are confused." The net result of this barnacle-like layering of international and domestic rules, standards, agreements, and criteria in the elimination of legal simplicity and the substitution in its stead of complexity and commercial uncertainty, which manifestly can not inure to the efficient and economical flow of world trade. All this makes a strong case for universal ratification of the Montreal Convention, which will supersede the Warsaw Convention and its various reformulations. Now that the Montreal Convention has entered into force, the insurance community may press the airlines to embrace it, which in turn may encourage the world's governments to ratify it. Under the Montreal Convention, the common law defence is available to the carrier even when it was not the sole cause of the loss or damage, again making way for the application of comparative fault principle. Hopefully, the recent entry into force of the Montreal Convention of 1999 will re-establish the international legal uniformity the Warsaw Convention of 1929 sought to achieve, though far a transitional period at least, the courts of different nations will be applying different legal regimes.

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Electron Microscopic Studies on Olfactory Bulbs in the Vertebrates by Phylogenetics (계통발생에 따른 척추동물의 뇌후구에 대한 전자현미경적 연구)

  • Choi, W.B.;Chung, Y.H.;Seo, J.E.
    • Applied Microscopy
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    • v.15 no.2
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    • pp.31-68
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    • 1985
  • Authors are trying to unveil the ultrastructural organization of olfactory bulb, which has been summerized under light microscopic level or communicated only in some detail in different view point until now. For the critical point of view, since the phylogenetical approach will give the ultimate value in the correlative study between structural and functional bases (Brodal, 1969), the present study was carried out light and electron microscopic analyses of the structures of the neurons and synaptic organizations in olfactory bulbs from different animals in phylogenetical scale. We selected each one species from five animal classes: the house rabbit(Oryctolagus cuniculus var. domesticus [Gmelin]) from Mammalia, the domestic fowl (Gallus gallus domesticus Brisson) from Aves, the viper (Agkistrodon hylys [G.P. Pallas]) from Reptilia, a frog (Bombiana orientalis Boulenger) from Amphibia and the crussian carp (Carassius carassius [Linne]) from Pisces. For light microscopic study, samples were fixed in 10% formalin and paraffin sections were stained with hematoxylin-eosin. For the electron microscopic study, the tissues were fixed by perfusion through the heart or immersion with 1% paraform-aldehyde-glutaraldehyde mixture (phosphate buffer, pH 7.4), and final tissue block trimmed under dissecting microscope were osmicated (1% OsO4), they were embedded in Araldite or Epon 812, and ultrathin sections were made by LKB-V ultratome following the inspection of semi-thin sections stained with toluidine blue-borax solution. Ultra-thin sections contrasted with uranyl acetate and lead citrate were observed with JEM 100CX electron microscope. We have summerized our morphological analyses as follows: 1. The olfactory bulb of rabbit, viper and frog shows the eight layers of fila olfactoria, glomerular, external granular, external plexiform, mitral cell, internal plexiform, internal granular, medullary but domestic fowl shows the five layers of glomerular, fibrillar, mitral, granular and medullary and the three layers of fibrilla, glomerular and medullary in crussian carp. The sharpness of demarcation between the layers shows deferential tendency according to phylogenetical order. 2. Mitral cells of vertebrate have large triangular or oval shape with spherical nuclei which contain not so much chromatin. The cytoplasm contains numerous cell organelles, of which Nissl's bodies or granular endoplasmic reticula arranged as parallel strands. Development of granular endoplasmic reticula were declined as the phylogentical grade is going lower. 3. Tufted cells of all animal are mostly spindle or polygonal contour and contain oval nuclei which located in periphery of cytoplasm. The nuclei of rabbit, fowl, viper and frog has relatively space chromatin, but a nucleus of crussian carp contain irregularly aggregated chromatin in karyoplasm. Their cytoplasmic volume and cell organelle contents are in between those of mitral cell and granular cell. They contain moderate amount of mitochondria, granular endoplasmic reticula, a few Golgi complex, polysomes, lysosome, etc. 4. Granule of cells of all the vertebrate amimals studied exhibit similar features; cells and their dense nuclei show spherical or oval contour, and they have the thin rim of cytoplasm which contain only a few cell organelles. 5. In rabbit, the soma of mitral cells were in contact with boutons with two types of synaptic vesicles, that is, round and flat vesicles, especially flat vesicles in boutons were showing reciprocal synapses. However, in domestic fowls, vipers, frogs and crussian carps, there were found boutons showing only spherical synaptic vesicles. 6. The boutons containing round synaptic vesicles were made contact with the some of tufted cell of olfactory bulb in the rabbits, fowls, vipers and frogs, but no synaptic boutons were observed in soma of tufted cells in crussian carps. In the frogs, there were observed dendrites were contact with the soma of tufted cells. 7. In the neuropils of plexiform, granular and glomerular layers olfactory bulbs in the vertebrate, the synapses were axo-large dendrites, axo-median and small dendrites, dendrodendritic, and axo-axonal contacts. However, in the neuropil of crussian carps, synapses were observed only in glomerular layer.

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Studies on the ecological variations of rice plant under the different seasonal cultures -II. A study on the year variations and prediction of heading dates of paddy rice under the different seasonal cultures- (재배시기 이동에 의한 수도의 생태변이에 관한 연구 -II. 재배시기 이동에 의한 수도출수기의 년차간변이와 그 조기예측-)

  • Hyun-Ok Choi
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.3
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    • pp.41-48
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    • 1965
  • This study was aimed at knowing the magnitude of year variation in rice heading dates under the different seasonal cultures, and to estimate the heading date in advance. Using six rice varieties such as Kwansan, Suwon#82, Suwon #144, Norin#17, Yukoo#132 and Paltal, the early, ordinary and late seasonal cultures had been carried out at Paddy Crop Division, Crop Experiment Station at Suwon for the six-year period 1959 to 1964. In addition the data of the standard rice cultures at the Provincial Offices of Rural Development for the 12-year period 1953 to 1954, were analyzed for the purpose of clarifying a relationship between variation of rice heading dates and some of meteorological data related to the locations and years. The results of this study are as follows: 1. Year variation of rice heading dates was as high as 14 to 21 days in the early seasonal culture and 7 to 14 days in the ordinary seasonal culture, while as low as one to seven days in the late seasonal culture which was the lowest among three cultures. The magnitude of variation depended greatly on variety, cultural season and location. 2. It was found out that there was a close negative correlation between the accumulated average air temperature for 40 days from 31 days after seeding and number of days to heading in the early seasonal culture. Accordingly, it was considered possible to predict the rice heading date through calculation of the accumulated average air temperature for the above period and then the linear regression(Y=a+bx). On the other hand, an estimation of the heading date in the late seasonal culture requires for the further studies. In the ordinary seasonal culture, no significant correlation between the accumulated average air temperature and number of days to heading was obtained in the six-year experiments conducted at Suwon. There was a varietal difference in relationship between the accumulated average air temperature for 70 days from seeding and number of days to heading in the standard cultures at the provincial offices of rural development. Some of varieties showed a significant correlation between two factors while the others didn't show any significant correlation. However, there was no regional difference in this relationship.

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Effects of Spent Composts of Se-Enriched Mushrooms on Carcass Characteristics, Plasma GSH-Px Activity, and Se Deposition in Finishing Hanwoo Steers (셀레늄급원으로 셀레늄강화버섯 폐배지의 급여가 거세한우의 도체특성, 혈중 GSH-Px활성 및 조직내 셀레늄축적에 미치는 영향)

  • Lee, S.H.;Park, B.Y.;Kim, W.Y.
    • Journal of Animal Science and Technology
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    • v.46 no.5
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    • pp.799-810
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    • 2004
  • This study was conducted to determine effects of spent composts of Se-enriched mushrooms (Se-SMC) as the dietary selenium source on carcass characteristics, plasma glutathione peroxidase(GSH-Px) activity and Se deposition in finishing Hanwoo steers. In combination with both Se-SMC and normal SMC, experimental treatment diets were formulated to contain 0.1, 0.3, 0.6 and 0.9 ppm of Se on a dry matter basis. A total of 20 finishing Hanwoo steers (average BW = 613 kg, average age = 20 to 24 mo) were allotted to treatments in four groups of five steers per pen for 12 wk preceding slaughter. While the experiment is employed, blood samples were taken to analyze Se concentration and GSH-Px activity, and muscle and liver samples were collected for analyses of Se contents in their tissues after slaughter. DMl and BW gain were not affected by dietary Se level and any toxic symptoms in treatments with a higher level of Se were not observed. No differences were noted for carcass characteristics. Se concentration in whole blood and plasma GSH-Px activity were linearly increased with the increasing level of dietary Se (P < 0.01). Se content in the hind leg for Se-SMC supplemented groups significantly increased (P < 0.05) upon dietary Se level, with 0.27, 0.37, 0.40 and 0.46 !1g1g dry, respectively. However, Se content in the loin was not affected by dietary Se levels. Se content in the liver was significantly increased(P < 0.05) as dietary Se increased, with 0.79, 1.40, 2.39 and 3.10 !1g1g dry, respectively. These results suggested that Se in the Se-SMC was highly bioavailable, and Se-SMC might be used not only as an inexpensive way of providing Se for ruminants but also as another way of producing Se-fortified beef.

Effects of Castration and Slaughtering Ages on Physico-chemical Characteristics of Hanwoo M. Longissimus dorsi (거세 및 도축시기가 한우 등심의 이화학적 특성에 미치는 영향)

  • Lee, K.C.;Par, N.H.;Jeong, J.;Lee, S.S.;Oh, Y.S.;Baek, K.H.;Jung, K.K.;Choi, C.B.
    • Journal of Animal Science and Technology
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    • v.46 no.2
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    • pp.165-172
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    • 2004
  • This study was conducted to investigate changes in physico-chemical characteristics of Hanwoo carcass according to different castration and slaughtering ages. Total 75 Hanwoo calves(average 4 months old) were randomly assigned to Control(bulIs). Treatment I. Il , III. or IV(castrated on 4. 9. 12. and 16 months of age, respectively) and slaughtered at 7. 9. 12, 16, and 28 months of age. Moisture contents in M. Longissimus dorsi of both Hanwoo bulls and steers were decreased as the slaughtering age increased. Moisture contents in all steer groups were significantly(P <0.05) lower(average 68.63%) than that in bulls(72.18%) at 28 months of age. Crude fat contents tended to increase as the slaughtering age increased. Crude fat contents in Treatment III(castrated at 12 months of age) at 28 months of age were 10.24% which was the highest among all steer groups. Crude protein contents in M. Longissimus dorsi of Hanwoo bulls were around 23% regardless of slaughtering age. Water holding capacity(WHC) of both bulls and steers tended to increase as the animals aged. Steers showed rapid increase in WHC after 16 months of age and showed higher(78.39%)) WHC than bulls(70.00%) at 28 months of age. Palmitic acid(CI8: 0) contents in Hanwoo bulls were increased as the slaughtering age increased(20.92 \longrightarrow 24.22%) whereas stearic acid(CI8: 0) contents were decreased(I8.l5 \longrightarrow11.32%). Oleic acid(CI8: I). a major unsaturated fatty acid. significantly(P <0.05) increased(25.59 \longrightarrow 37.90%) when slaughtering age of Hanwoo increased. Changes in fatty acid composition in Hanwoo steers were similar to those in bulls except oleic acid contents were higher and stearic acid contents were lower than bulls at 28 months of age.