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Studies on Prevalence of Meat Allergy and Potential Beef Allergens (식육알레르기 발생 경향 및 잠재적 우육알레르겐에 관한 연구)

  • Jeong, Bo-Young;Kim, Dong-Yeop;Fan, Jiang Ping;Chung, Hyun-Chae;Han, Gi-Dong
    • Food Science of Animal Resources
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    • v.29 no.2
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    • pp.151-156
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    • 2009
  • The prevalence of food allergies was investigated using questionnaires with 300 subjects whose ages ranged from 19 to 24 years old and the causative food allergens was analyzed using immunological analysis with serum of the subjects who answered that they have/had food allergy. The questionnaire showed that 11.33% of subjects have/had experience of food hypersensitivity, where the main causative foods were fish, beef, chicken, milk, egg, and pork in order. The meat allergy shared 4.65% (2.33% for beef, 1.66% for chicken, 0.66% for pork) in the prevalence of food allergies. The causative beef allergens were investigated with the serum of 6 subjects who have had beef allergy. Western blots were carried out with the serum of P6 subject who showed a positive reaction to beef extract in ELISA. The two specific bands were detected in beef extract on the PVDF membrane, and no band was detected in extracts of pork and chicken. A calculation of the distance of migration by SDS-PAGE enabled the molecular masses of the two bands to be estimated as 67kDa and 31kDa, respectively. The 67kDa was revealed as bovine serum albumin (BSA) which is one of the important beef allergens as reported previously though an analysis of the N-terminal amino acid sequence. However we could not identify the sequence of 31kDa, probably because they comprised several subunits and were modified proteins such as glycoprotein that were unlikely to be easily degraded by the Edman method. The 31kDa band were dyed with the PAS (periodic acid-schiff reagent), suggesting that it might be a glycoprotein. These results suggested that the 31kDa might be considered as a novel potential beef allergen which is not reported previously, although further studies are needed.

Adiponectin Gene Cloning and Its Expression in Insect Cell Expression System

  • Yuh, In Suh;Sheffield, Lewis G.
    • Reproductive and Developmental Biology
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    • v.36 no.3
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    • pp.193-198
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    • 2012
  • This study was to examine expression of the recombinant full-length adiponectin (recombinant adiponectin) in insect ovarian cell culture system and to characterize structural properties of the recombinant adiponectin secreted in medium. Gene construct encoding the recombinant adiponectin contained N-terminal collagen-like domain (110 Amino Acids, AAs), C-terminal globular domain (137 AAs) and C-terminal peptides for detection with V5 antibody (26 AAs included adaptor peptide) and purification using the 6xHis tag (6 AAs). The approximate molecular weight of the product (monomer) was 35 kDa. Molecular mass species of the expressed recombinant adiponectin were monomer (~35 kDa), dimer (~70 kDa), trimer (~105 kDa) and hexamer (~210 kDa). The major secreted species were the LMW forms, such as monomer, dimer, and trimer. There was MMW of hexamer as minor form. HMW multimers (~300 kDa) were shown as a tracer or not detected on the SDS-PAGE in several experiments (data not shown). The multimer forms in this study were not compatible to those in animal or human serum and adipose tissue by other researcher's study in which the major multimer forms were HMW. By protein denaturing experiments with reducing reagent (${\beta}$-MeOH), anionic detergent (SDS) and heat ($95^{\circ}C$) on the SDS-PAGE, not all adiponectin multimers seemed to have disulfide bond linked structure to form multimers. The recombinant adiponectin which expressed in insect ovarian cell culture system seemed to have the limitation as full physiological regulator for the application to animal and human study.

Comparison of Biochemical and Immunological Properties Between Rat and Nicotiana glutinosa Ornithine Decarboxylase

  • Lee, Yong-Sun;Cho, Young-Dong
    • BMB Reports
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    • v.34 no.5
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    • pp.408-414
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    • 2001
  • Ornithine decarboxylase (EC 4.1.1.17) is an essential enzyme for polyamine synthesis and growth in mammalian cells and plants. We compared the biochemical and immunological properties of rat and Nicotiana glutinosa ODC by cloning and expressing the recombinant proteins. The primary amino acid sequence between rat and N. glutinosa ODC had a 40% homology The molecular weight of the overexpressed rat ODC was 53 kDa, and that of N. glutinosa was 46.5 kDa. Adding 1 mM of putrescine to the enzyme reaction mixture inhibited both rat and N. glutinosa ODC activity to 30%. Agmatine had an inhibitory effect only on N. glutinosa ODC. Cysteine and lysine modifying reagents reduced both ODC activities, verifying the key roles of cysteine and lysine residues in the catalytic mechanism of ODC. ELISA was performed to characterize the immunological difference between the rat and plant ODC. Both the rat and N. glutinosa ODC were recognized by the polyclonal antibody that was raised against purified N. glutinosa ODC, but the rat ODC was 50-fold less sensitive to the antibody binding. These results indicate that even though both ODCs have the same evolutionary origin, there seems to be a structural distinction between the species.

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The Effect of Fatigue Crack Behavior on the Variable Depth of Micro Hole Defects in SM20C at the Symmetric Position (대칭위치에 존재하는 미소원공결함의 깊이변화가 SM20C의 피로균열거동에 미치는 영향)

  • 송삼홍;김성태
    • Proceedings of the Korean Society of Precision Engineering Conference
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    • 2002.05a
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    • pp.856-860
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    • 2002
  • The main objective of this study is to consider the effect of fatigue crack behavior on the variable depth of micro hole defects in SM20C at the symmetric position. The fatigue crack propagation test is performed by rotary bending fatigue test machine. The relationship between crack length(2a), cycles(N) and crack growth rate(da/dN) are investigated in this study. The result from the rotary bending fatigue test under the applied stress at 250MPa turned out that the fatigue life illustrated almost constant when the depth of symmetric micro hole deflects is both part A and B at the hope depth(h) = 0.5mm.

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Fracture Mechanic's Approach on Retardation Behaviors under Overloading (과대 하중작용 시 균열성장 지연 거동에 대한 파괴역학적 정리)

  • Kang, Yong-Goo;Park, Jun-Young
    • Journal of the Korean Society of Manufacturing Process Engineers
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    • v.11 no.5
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    • pp.13-20
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    • 2012
  • In order to clarify the effect of overload on crack growth behaviors, fatigue tests for overload were carried out for round plain specimens of SM45C steel. In the experiment, typical semi-elliptical crack shape was found and further crack growth behaviors were tested. Using three types of single overload fatigue tests, Crack growth retardation phenomenon were examined. The growth rate of surface crack(da/dN) during retardation period was analyzed in terms of ${\Delta}K$ and ${\Delta}K_{eff}$. On the growth rate of surface crack analyzed by ${\Delta}K$, the dependence of overload stress levels appears. However, on the growth rate by ${\Delta}K_{eff}$ obtained by Willenborg analysis, there is a non-liner relationship between da/dN and ${\Delta}K_{eff}$ with narrow scatter band.

Fracture Mechanic's Approach on Retardation Behaviors under Overloading (과대하중작용 시 균열성장 지연거동에 대한 파괴역학적 정리)

  • Kang, Yong-Goo;Park, Jun-Young
    • Journal of the Korean Society of Manufacturing Process Engineers
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    • v.11 no.4
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    • pp.82-89
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    • 2012
  • In order to clarify the effect of overload on crack growth behaviors, fatigue tests for overload were carried out for round plain specimens of SM45C steel. In the experiment, typical semi-elliptical crack shape was found and further crack growth behaviors were tested. Using three types of single overload fatigue tests, Crack growth retardation phenomenon were examined. The growth rate of surface crack(da/dN) during retardation period was analyzed in terms of ${\Delta}K$ and ${\Delta}K_{eff}$. On the growth rate of surface crack analyzed by ${\Delta}K$, the dependence of overload stress levels appears. However, on the growth rate by ${\Delta}K_{eff}$ obtained by Willenborg analysis, there is a liner relationship between da/dN and ${\Delta}K_{eff}$ with narrow scatter band.

Physicochemical Properties and Biological Activities of Collagens with Different Molecular Weights from Alaska Pollack (Theragra chalcogramma) Skin (명태 껍질 유래 콜라겐의 분자량에 따른 이화학적 특성 및 생리활성)

  • Yang, Su-Jin;Hong, Joo-Heon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.10
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    • pp.1535-1542
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    • 2014
  • This study was conducted to investigate the physicochemical properties and biological activities of collagens with different molecular weights from Alaska pollack (Theragra chalcogramma) skin as well as their efficacies as functional materials. The molecular weights of collagens were between 1~10 kDa (below 1 kDa (AP1), 1~3 kDa (AP2), 3~10 kDa (AP3), and above 10 kDa (AP4). The protein content of AP4 (40.19 g/100 g) was the highest. Collagen contents of AP1, AP2, AP3, and AP4 were 36.43, 32.23, 19.23, and 14.89%, respectively. The free amino acid and essential amino acid contents of AP1 were higher than those of AP2, AP3, and AP4. Fourier transform infrared spectroscopy spectra of collagens with different molecular weights showed wavenumbers representing the regions of amide I, amide II, amide III, and amide A, respectively. The electron-donating ability (29.51%) and SOD-like activity (38.45%) of AP1 were higher than those of AP2, AP3, and AP4. Tyrosinase inhibition activity of AP1 improved with higher treatment concentration. The rate of inhibition of MMP-1 production in HS68 cells exposed to UVB was suppressed by treatment with AP1 (29.78%) and AP2 (26.49%) at 1 mg/mL. Furthermore, there was a strong correlation between DPPH, superoxide dismutase, tyrosinase activity, and MMP-1 inhibition rate of collagens with different molecular weights.

Changes of the Extracellular Concentrations of Striatal Dopamine and Its Metabolites by MK-801 in Morphine-Dependent Rats (MK-801 투여에 의한 몰핀의존성랫드 뇌선초체중 도파민신경절달물질의 변화)

  • 이선희;신대섭;유영아;류승렬;김대병
    • Biomolecules & Therapeutics
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    • v.6 no.1
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    • pp.25-30
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    • 1998
  • The roles of dopamine(DA) and N-methyl-D-aspartate(NMDA) system in the development and expression of morphine dependence were investigated by monitoring the concentrations of extracellular DA and its metabolites by in vivo microdialysis and simultaneous observation of behavioral changes in morphine dependent rats. Extracellular DA level in caudate putamen of morphine-dependent rat was decreased and the concentrations of its metabolites, dihydroxy phenylacetic acid(DOPAC) and homovanillic acid(HVA), were increased during naloxone-precipitated withdrawal. DA contents were recovered to normal levels by pretreatment of MK-801, a noncompetitive NMDA receptor antagonist, which may explain the mechanism of diminishing effect of MK-801 on withdrawal symptoms in morphine-dependent rats. MK-801(0.3 mg/tg, i.p.) induced the untoward hamful neurological signs such as ataxia and severe rotations, which may be produced by hyperactivation of dopaminergic system. These results suggest that MK-801 may inhibit the expression of mophine dependence by altering the dopamine release.

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NONCOMPETITIVE NMDA RECEPTOR ANTAGONISTS INHIBIT APOMORPHINE-INDUCED CLIMBING BEHAVIOR IN RESERPINE-TREATED MICE

  • Kim, Hack-Seang;Rhee, Gyu-Seek;Park, Woo-Kyu
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1996.04a
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    • pp.247-247
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    • 1996
  • Previous work in our laboratory has shown that noncompetitive N-methyl-D-aspartate (NMDA) receptor antagonists, MK-801, ketamine, dextrorphan and dextromethorphan cause a pronounced inhibition of apomorphine-induced cage climbing behavior in intact mice, suggesting the involvement of NMDA receptors in the glutamatergic modulation of dopaminergic function at the postsynaptic dopamine (DA) receptors: Therefore, in order to definitively establish the involvement of NMDA receptor in the apomorphine-induced dopaminergic response at the postsynaptic DA receptor, it is necessary to investigate whether or not the noncompetitive NMDA receptor antagonists would inhibit these phenomena not only in intact mice but also in the mice that are devoid of any involvement of indirect dopaminergic function. To minimize the risk of any indirect involvement of NMDA antagonists with DA neurons, vesicular DA stores were first depleted with reserpine.

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