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Development of Baccillus megaterium Disk Assay Kit for the Determination of Antibacterial Residues in Animal Tissues (식육중 잔류 향균물질의 검출을 위한 Bacillus megaterium 디스크 검사킷트 개발)

  • 손성완;조병훈;진남섭;이혜숙;윤순학;김재학;이재진;이영순
    • Journal of Food Hygiene and Safety
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    • v.11 no.4
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    • pp.315-321
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    • 1996
  • Various antimicrobial drug screen tests have been used in order to ensure food safety. However, the conventional screen tests, the Swab Test on Premises(STOP, USA), the Calf Antibiotic and Sulfa Test(CAST, USA) and the European Economic Community 4-plate Test(FPT, EU) are not sufficiently rapid or sensitive enough to detect low levels of sulfa drugs in meat. We developed a new screen test kit for the determination of the antimicrobial residues in meat called the Bacillus megaterium Disk Assay(BmDA). A comparison of BmDA with the older screen tests showed BmDA was as good as the older ones with several advantages. The new test kit is faster-it can be read in 4∼6 hours instead of 16∼18 hours. Moreover, BmDA can discriminate sulfa drugs from other antimicrobial drugs because p-aminobenzoic acid countacts the inhibiting action of sulfa drugs. Minimum detectable levels of sulfa drugs were significantly improved at the lever of 0.025*0.1 pp, compared with the level of 1.0 ppm in FPT. A comparison of BmDA with the older screen tests in HPLC confirmed meat samples exceeded the Korean tolerance value of 0.1 ppm showed BmDA was the most sensitive in the microbiological screen tests. As the microbiological screen tests have already known, a person familiar with simple laboratory techniques should have no difficulty in using it to detect antimicrobial residues in meat. This would be a simple, economic method of antimicrobial residues detection which might be succesfully used by many laboratories.

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Protein Profile of Embryonic Stem Cells Following Differentiation

  • D. I. Jin;Kang, H. S;Kim, H. J.;Lee, S. H.;Park, C. S.;K. S. Im;Lee, H. M.
    • Korean Journal of Animal Reproduction
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    • v.26 no.4
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    • pp.377-384
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    • 2002
  • To investigate the expression patterns of proteins and growth factor signals in differentiated rabbit embryonic stem (ES) cells, ES cells with confluent stage grown of feeder layer and differentiated cells into embryoid bodies (EB) without feeder cell were applied to protein gel and Western blotting analysis. There were 66kDa and 28kDa specifically expressed in differentiated ES cell but not in undifferentiated ES cell while 25kDa protein band showed up in only undifferentiated ES cells. Also there were some difference of protein bands in several area of gel between differentiated and undifferentiated ES cells such as about 100 kDa, 50kDa and 27kDa areas, but there was no difference in band pattern of one-dimensional gel analysis between mouse ES cells and rabbit ES cells. IGF-I receptor and EGF receptor were expressed in differentiated cells and undifferentiated cells. And ICF-I and EGF were not expressed in both differentiated and undifferentiated cells. These results indicated that ES cells express their own proteins to inhibit differentiation while EB cells synthesize different proteins to differentiate, and 16F-I receptor and EGF receptor were expressed in both ES and EB cells probably for the different functions.

$\alpha$- and $\beta$-Amylase Isozyme Expresser Native Proteins in Tropical Silkworm Bombyx mori L.

  • Chattopadhyay, G.K.;Verma, A.K.;Sengupta, A.K.;Das, S.K.;Urs, S.Raje
    • International Journal of Industrial Entomology and Biomaterials
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    • v.8 no.2
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    • pp.189-194
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    • 2004
  • Amylase isozyme based three multivoltine viz., N+p, Np, N+ $p^{cho}$ and two bivoltine-D6+p, D6p syngenic lines (Syn. L) were developed from germplasm (GP) stocks Nistari (N) and D6 respectively. haemolymph isozyme pattern at pH 7.0 and 8.5 depicted a total 11 number (Am $y_{1 to 6}$ at pH 7.0 and Am $y^{l to 5}$ at pH 8.5) of native proteins (NP) of various sizes are amylase isozyme expressers. Among eleven NPs, two NPs of 770 kDa (Am $y^{6}$ at pH 7.0) and 376 kDa (Am $y^3$ at pH 8.5) are $\alpha$-amylase expressers and remaining NPs of 370, 364, 350, 329 and 274 kDa at pH 7.0 and 206, 292, 416, 725 kDa at pH 8.5 are $\beta$-amylase expressers. Accordingly, digestive juice amylase isozyme pattern at aforesaid pH also depicted a total number of 10 NPs (Am $y^{1 to 5}$) at each pH 7.0 and 8.5 are amylase expressers of which NP of 387 kDa (Am $y^4$ at pH 7.0) and 780 kDa (Am $y^{5}$ at pH 8.5) are a-amylase expresser. Remaining NPs of 338,297 & 216 kDa at pH 7.0 and 370, 341, 329 &302 kDa at pH 8.5 are $\beta$-amylase expresser. Recurrent backcross lines (RBL) viz., N+pRBL and NpRBL were developed through introgression of high shell weight character (a multigenic trait) to be used further for congenic line (Con. L) development and to understand any association with introgressed character. Isozyme pattern in haemolymph of RBLs depicted only one $\alpha$-amylase of 770 kDa at pH 7.0 and 376 kDa at pH 8.0 with three and four respective $\beta$-amylase bands but in bivoltine lines numbers of $\beta$-amylase bands vary between 1 to 2 at aforesaid pH. Variability was also observed in digestive juice of multivolitine and its RBLs but bivoltine lines express null activity at both pH except appearance of one very week $\alpha$-amylase band D6+p at pH 8.5. Overall study suggests that not a single NP at both pH is common for expression of any band of amylase isozyme i.e., a totally different set of proteins are the amylase isozyme expresser at specific pH and no molecular factor of amylase is associated in developed RBLs which showed improvement on survival, single cocoon shell weight (SCSW) and single filament length over receptor parents.s.s.s.

Physicochemical and Sensory Properties of Water Soluble Chitosan (수용성 Chitosan의 이화학적 및 관능적특성)

  • Kim, Dong-Ho;Lee, Chan;Kim, Kwang-Ok;Lee, Young-Chun
    • Korean Journal of Food Science and Technology
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    • v.31 no.1
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    • pp.83-90
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    • 1999
  • Although the chitosan has many functional properties due to its cationic amino groups, the application of chitosan in foods is limited by its poor water solubility, bitter taste and astringency. This study was conducted to investigate physicochemical and sensory properties of chitosan hydrolysates in various molecular weights obtained by ultrafiltration after enzymatic hydrolysis. As molecular weight decreased, the solubility of chitosan hydrolysates increased, while the viscosity and emulsion stability decreased. High molecular weight chitosan hydrolysates (>30 kDa) exhibited 800% of fat binding capacity, while low molecular weight ones $(3{\sim}30\;kDa)$ showed 500% of fat binding capacity. Water soluble chitosan hydrolysates exhibited no color differences. Freeze-thaw stability of chitosan hydrolysates was good, without variations among fractions. Cholesterol binding capacity of chitosan hydrolysates was changed from 24% to 36% with increasing molecular weights. From sensory evaluation of chitosan hydrolysates, it was found that bitterness, astringency, chemical flavor and fish flavor of chitosan hydrolysates were very weak.

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Embryotoxic effects of DA-125, a new anthracycline anticancer agent, in rats (새로운 안트라사이클린계 항암제 DA-125의 랫트에 있어서 태자독성효과)

  • Chung, Moon-koo;Kim, Jong-choon
    • Korean Journal of Veterinary Research
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    • v.34 no.1
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    • pp.165-172
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    • 1994
  • DA-125 is a new anthracycline antitumor antibiotic, which is derived from adriamycin. The potential of DA-125 to induce embryotoxicity was evaluated in the Sprague-Dawley rats. One hundred twenty naturally mated SD rats(sperm in vaginal lavage=day 0) were distributed among three treated groups and a control group. DA-125 was administered intravenously at dose levels of 0. 0.1, 0.3 and 1.0mg/ kg/day. Dams were treated from day 7 to 17 of gestation and were subjected to the caesarean section on day 20. At 1 mg/kg, reduced food intake, reduced body weight and decreased weight of spleen were observed in dams. An increase in the resorption rate and a reduction in the fetal weight were also found. In addition, various types of external, visceral and skeletal malformations occurred at an incidence of 11.9, 41.8 and 14.5%, respectively. Characteristic malformations include exencephalia, gastroschisis, cleft lip, dilatation of lateral and 3rd ventricle, fused ribs, among others. There were no signs of maternal toxicity or embryotoxicity at 0.1 and 0.3mg/kg. The results show that the test agent DA-125 is embryotoxic at maternally subtoxic dose in rats.

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Comparison of Allergens in Genetically Modified Soybean with Conventional Soybean (유전자변형 콩과 자연 콩의 알레르기 유발원 비교)

  • 박재현;정승태;김재희;김지영;노건웅
    • YAKHAK HOEJI
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    • v.45 no.3
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    • pp.293-301
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    • 2001
  • Genetically modified organism (GMO) using recombinant DNA technique has been exponentially increased, however there are still arguments for the safety of GM foods. The objective of this research was to compare the allergens of GM soybean(Roundup Ready$^{TM}$) with conventional soybeans. Each soybean extracts were prepared as crude extracts, heated extracts, and as heated and simulated gastric quid (SGF)-digested samples to characterize the stability of allergens to physicochemical treatment. Positive sera from 20 soybean-sensitive patients and control sera from 5 normal subjects were used to identify the endogenous allergens in soybeans. Specific-IgE binding activities to each soybean preparations were evaluated by ELISA and immunoblot technique. In ELISA result, IgE binding activities of positive sera to soy crude extracts generally showed two fold higher mean value than those of control sera, how-ever there was no significant difference between GM soybean and natural soybean varieties. Extracted proteins form each of the soybean preparations were separated with SDS-PAGE. The band pattern of GM soybean was very similar to those of natural soybean varieties. Immunoblots for the different soybeans revealed no differences in IgE-binding protein patterns, moreover, disclosed five prominent IgE-binding bands (75, 70, 50, 44 and 34 kDa) in crude extracts, four (75, 70, 44 and 34 kDa) in heated preparations, one (50 kDa) in heated and SGF-digested preparations. These IgE binding bands were consistent with previously reported results on the soybean. These results indicate that GM soybean (Roundup Ready$^{TM}$) is no different from natural soybean in terms of its allergen.gen.

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Comparison of Biochemical Characterization of Korean and Chinese Mung Bean Lectin (한국산 녹두와 중국산 녹두에 있어서 Lectin의 생화학적 특성 비교)

  • Roh, Kwang Soo
    • Journal of Life Science
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    • v.24 no.6
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    • pp.603-611
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    • 2014
  • The lectins were separated from Korean and Chinese mung bean seeds finally via chromatography using Sephadex G-100 and their biochemical features were studied and compared. They showed no hemagglutination with human red blood cells regardless of trypsin treatment and showed hemagglutination with only trypsin treated rabbit red blood cells. The molecular weights of two lectins were identified as 54 kDa and 28 kDa by SDS-PAGE. It was found that while the optimal reaction temperature of the lectin from Korean mung bean was $60^{\circ}C$, that of the lectin from Chinese mung bean seeds was $50^{\circ}C$. It was found also that the most thermal stable temperature of the seed lectin from Korean mung bean seeds was $50^{\circ}C$ and the lectin from Chinese mung bean was $40-50^{\circ}C$. The lectin from Korean mung bean seeds showed the highest activity at pH 3.2 and the lectin from Chinese mung bean showed the highest activity at pH 6.2. It was identified that when treating a denaturant, thiourea and guanidine-HCl resulted in no hemagglutination, so they induced denaturalization. It was identified also that there was no hemagglutination with urea, so it did not induced denaturalization. They showed no septicity to 6 types of carbohydrates including D-glucose. In addition, the lectins from the two mung bean seed had specificity to metal ions.

Characterization of Bacillus stearothermophilue Cyclodextrin Glucanotransferase that Expressed by Saccharomyces cerevisiae (Saccharomyces cerevisiae에서 발현된 Bacillus stearothermophilus Cyclodextrin Glucanotransferase의 특성)

  • 박현이;전숭종;권현주;남수완;김한우;김광현;김병우
    • Microbiology and Biotechnology Letters
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    • v.30 no.4
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    • pp.293-297
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    • 2002
  • The cyclodextrin glucanotransferase (CGTase) gene from Bacillus stearothermophilus NO2 was expressed in Saccharomyces cerevisiae 2805 under the adhl promoter. The CGTase was purified from S. cerevisiae 2805/pVT-CGTS. The purified enzyme exhibited a optima of activity around pH 7.0 and $65^{\circ}C$. Thermal stability of the enzyme was increased fairly as compared with the CGTase of B. stearothermophilus NO2. The conversion yield of cyclodextrin (CD) and the production ratio of $\alpha$-, $\beta$,-, ${\gamma}$-CD from starch were showed similarly aspect to the CGTase of B. stearothermophilus NO2.