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High-Level Expression of A Bacillus subtilis Mannanase Gene in Escherichia coli. (대장균에서 Bacillus subtilis의 Mannanase 유전자 과잉발현)

  • 권민아;손지영;윤기홍
    • Microbiology and Biotechnology Letters
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    • v.32 no.3
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    • pp.212-217
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    • 2004
  • The gene coding for mannanase from Bacillus subtilis WL-7, a number of glycosyl hydrolase family 26, was hyperexpressed in Escherichia coli. Two recombinant plasmids, pE7MAN and pENS7, were constructed by introducing the complete mannanase gene and the mature mannanase gene lacking N-terminal signal peptide region into a expression vector pET24a(+), respectively. The level of mannanase produced by E. coli BL21 (DE3) carrying pENS7, which included the mature mannanase gene, was considerably higher than that by E. coli BL21 (DE3)/pE7MAN. Almost mannanase produced by the recombinant E. coli carrying pENS7 at growth temperature of $37^{\circ}C$ existed as inactive enzyme of insoluble form. Growth at temperature below $31^{\circ}C$ increased the soluble fraction of mannanase having catalytic activity in the recombinant E. coli cells. The highest productivity of active mannanase was observed in cell-free extract of the recombinant E. coli grown at growth temperature ranging from $25^{\circ}C$ to $28^{\circ}C$, while mannanase activity per soluble protein of the cell-free extract was highest in the cells grown at $^31{\circ}C$.

Cellular Prion Protein Enhances Drug Resistance of Colorectal Cancer Cells via Regulation of a Survival Signal Pathway

  • Lee, Jun Hee;Yun, Chul Won;Lee, Sang Hun
    • Biomolecules & Therapeutics
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    • v.26 no.3
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    • pp.313-321
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    • 2018
  • Anti-cancer drug resistance is a major problem in colorectal cancer (CRC) research. Although several studies have revealed the mechanism of cancer drug resistance, molecular targets for chemotherapeutic combinations remain elusive. To address this issue, we focused on the expression of cellular prion protein ($PrP^C$) in 5-FU-resistant CRC cells. In 5-FU-resistant CRC cells, $PrP^C$ expression is significantly increased, compared with that in normal CRC cells. In the presence of 5-FU, $PrP^C$ increased CRC cell survival and proliferation by maintaining the activation of the PI3K-Akt signaling pathway and the expression of cell cycle-associated proteins, including cyclin E, CDK2, cyclin D1, and CDK4. In addition, $PrP^C$ inhibited the activation of the stress-associated proteins p38, JNK, and p53. Moreover, after treatment of 5-FU-resistant CRC cells with 5-FU, silencing of $PrP^C$ triggered apoptosis via the activation of caspase-3. These results indicate that $PrP^C$ plays a key role in CRC drug resistance. The novel strategy of combining chemotherapy with $PrP^C$ targeting may yield efficacious treatments of colorectal cancer.

Nutritional Status and Indicators of Immune Function of Elderly Residing in Low Income Urban Area (도시생활보호노인들의 영양상태와 면역지표에 관한 연구)

  • 손숙미;윤혜영
    • Korean Journal of Community Nutrition
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    • v.2 no.4
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    • pp.547-555
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    • 1997
  • This study was conducted to assess the nutritional status of the elderly from low income areas and its relationship with the indices of immunity. Ninety nine(male : 28, female : 71) healthy, subjects aged over 65 years staying in homes in Seoul were assessed with regard to their dietary intake and biochemical measurements. Their mean daily energy intakes were 1256kcal and 109kcal for males and females respectively. Mean serum IgG were 2356mg/dl in men and 2161mg/dl for women, higher than the normal range. Mean serum $C_3$ were 120.9mg/dl and 130.6mg/dl for males and females respectively and mean $C_4$ were 30.6mg/dl and 31.9mg/dl. The mean $C_3$ and $C_4$ were within the normal range and the proportion of subjects whose $C_3$ and $C_4$ were at a higher level than the upper limit were 15.4$\%$ and 16.3$\%$ respectively. Complement $C_3$ showed significant correlation with serum albumin(p〈0.01), serum cholesterol(p〈0.00l, serum TG(p〈0.05), LDL-C(p$\ll$$\ll$$\ll$$\ll$$\ll$$\ll$$\ll$$\ll$$\ll$$\ll$$\ll$$\ll$$\ll$q0.001), VLDL-C(p$\ll$0.05), serum copper(p$\ll$0.05) and serum Ca(p$\ll$0.01). (Korean J Community Nutrition 2(4) : 547-555, 1997)

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Synthesis and Structure of Tetrahomodioxa p-phenylcalix(4)arene dihexylether (Tetrahomodioxa p-phenylcalix(4) arene dihexylether의 합성 및 구조에 관한 연구)

  • 노광현;박영자
    • Korean Journal of Crystallography
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    • v.13 no.3_4
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    • pp.158-164
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    • 2002
  • Tetrahomodioxa p-phenylcalix(4)arene dihexylether(C/sub66/H/sub68/O/sub6/) has been synthesized and structurally characterized by X-ray diffraction. Reaction of tetrahomodioxa p-phenylcalix(4)arene with hexyl halide and NaH in DMF leads to the dihexyl derivatives, 7,13,21,27-tetraphenyl-29,31-dihexyloxy- 2,3,16,17-tetrahomo-3,17-dioxacalit(4)arenes. The crystal is orthorhombic, P2₁2₁2₁, a= 9.764(2), b=16.167(2), c=32.994(3) Å, V=5208(1) Å, Z= 4, Dc = 1.221 gcm/sup -3/. The structure was solved by direct methods and refined by full-matrix least squares. Refinement converged at R = 0.070 for 2009 observed reflections. This molecule has a C-1,2-alternate conformation with pseudo-centrosymmetry and has two pairs of opposite phenyl rings, which are approximately parallel to each other. The benzene rings A and B are up, and the rings C and D rings are down with respect to the plane of the macrocyclic ring.

Carcass Characteristics, Chemical Composition and Fatty Acid Profile of Longissimus Muscle of Bulls and Steers Finished in a Pasture System Bulls and Steers Finished in Pasture Systems

  • Aricetti, Juliana Aparecida;Rotta, Polyana Pizzi;do Prado, Rodolpho Martin;Perotto, Daniel;Moletta, Jose Luiz;Matsushita, Makoto;do Prado, Ivanor Nunes
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.10
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    • pp.1441-1448
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    • 2008
  • This experiment was carried out to study the carcass characteristics, chemical composition and fatty acid profile of the Longissimus muscle (LM) of bulls (10) and steers (17) finished in a pasture system. Animals (1/2 Zebu vs. 1/2 Aberdeen Angus) were fed in a pasture system (Hermatria altissima) and with a supplement of soybean meal, cracked corn, urea, limestone and mineral salts, twice a day. Both animal groups were slaughtered at 27 months of age, with an average 508.88 kg of live weight. Final weight, hot carcass weight and texture were similar (p>0.05) between bulls and steers. Carcass dressing, fat thickness, color and marbling were higher (p<0.02) in steers. Conversely, the Longissimus area was greater (p<0.05) in bulls. Moisture levels were higher (p<0.01) in bulls. Ash, crude protein, total lipids and total cholesterol levels were higher (p<0.10) in steers. C14:0, C16:0, C16:1 n-7 and C18:1 n-9 fatty acids percentages were higher (p<0.06) in steers. C16-1 n-10, i 17:0, C19:0, C18:2 n-6, C18:3 n-3, C20:2 n-3 C20:4 n-6, C20:5 n-3, C22:1 n-7, C22:4 n-6 and C22:5 n-3 fatty acids percentages were higher (p<0.05) in bulls. The percentage of the other fatty acids was similar (p>0.05) in bulls and steers. The levels of monounsaturated fatty acids were higher (p<0.02) in steers while polyunsaturated fatty acids, n-3, n-6, PUFA/SFA and n6:n3 ratios, were higher (p<0.05) in bulls. The CLA contents (mg/g lipids) were similar (p>0.10) in bulls and steers.

Characterization of $SiO_2-P_2O_5-B_2O_3$ Glass Soot fabricated by Flame Hydrolysis Deposition (화염 가수분해 증착에 의해 형성된 $SiO_2-P_2O_5-B_2O_3$ 유리 미립자의 특성)

  • 최춘기;정명영;최태구
    • Journal of the Korean Ceramic Society
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    • v.34 no.8
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    • pp.811-816
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    • 1997
  • SiO2-P2O5-B2O3 glass soot was fabricated by flame hydrolysis deposition and their properties by SEM, XRD, TGA-DSC were investigated., The mechanism of consolidation process of a glass soot as a function of consolidation temperature was analyzed by SEM observations. In the XRD patterns, the crystalline peaks which seem to be generated from B2O3 and BPO4 were observed. When the temperature of heat treatment exceeded 105$0^{\circ}C$, the non-crystalline state of SiO2-P2O5-B2O3 glass was observed. In the TGA-DSC curves, the evaporation of water molecule by a sudden endothermic reaction was observed at 128$^{\circ}C$ and a broad endothermic peak was seen in the temperature range of 40$0^{\circ}C$-95$0^{\circ}C$, without any weight loss. Finally, this peak was began to recover its baseline at 953$^{\circ}C$. This point is equal to the temperature at which the densification begins. Furthermore, we observed that the addition of dopants such as P2O5 and B2O3 decrease the onset of consolidation temperature till 95$0^{\circ}C$.

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Production of Enzyme Modified cheese (Enzyme Modified Cheese 제조)

  • Seo, Hyeong-Ju;Son, Jong-Yeon;Kim, Yun-Suk
    • The Korean Journal of Food And Nutrition
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    • v.8 no.3
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    • pp.192-198
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    • 1995
  • For the production of EMC, various professes and lipases were used to hydrolyse cheese sulk. The optimal conditions of various proteases were as follows, pronase-3$0^{\circ}C$, p14 7.0, pancreatln-4$0^{\circ}C$, pH 8.0, pacific protease-3$0^{\circ}C$, pH 7.0 and protease from Asp. sp. -5$0^{\circ}C$, pH 8.0. The optimal conditions of various lipases were as follows ; pancreatic lipase-5$0^{\circ}C$, pH 8.0, palatase ML-5$0^{\circ}C$, pH 7.0 and lipase form Candida -4$0^{\circ}C$, pH U.0. After hydrolysation under optimal conditions, the amounts of free amino acid and free fatty ac14 were increased with reaction time. Hydrolysates of pacific protease and pronase were showed high amount of free amino acid(0.67mg/ml and 0.74mg/ml). Especially EMC had high amount of glutamic acid and leucine. Lipase from Candida cylindracea produced high amount of free fatty acid (24.63 mg/ml) Butyric acrid, palmitic acid, stearic acid and oleic acid among free fatty acids were showed high amounts. Sensory evaluation of various MC were tasted nth 8 panelist. EMC produced with pancreatic lipase was most bitterness and EMC produced with palatase ML was best acceptable cheese flavor.

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Studies on Nitroaniline derivative of 2, 2'-methylene-bis-(3, 4, 6-trichloroacetoxy benzene by Mannich reaction Part I. Synthesis and Antimicrobial activity (2, 2'-methylene-bis-(3, 4, 6-trichloroacetoxy benzene)의 nitroaniline 계 mannich bases에 관한 연구 I. 합성 및 항균성)

  • Yu, Ju-Hyun;Kim, Yu-Sam;Kim, Jong-Ho;Yang, Ryung
    • Korean Journal of Food Science and Technology
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    • v.6 no.1
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    • pp.6-11
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    • 1974
  • Four new compounds; 2,2'-methylene-bis [3, 4, 6-trichloro ${{\beta}-(o-nitroanilino)$ propionoxy} benzene]; m.p. $200{\sim}202^{\circ}C,\;C_{31}H_{22}O_8N_4Cl_6$ 2,2'-methylene-bis [3, 4, 6-trichloro ${{\beta}-(p-nitroanilino)$ propionoxy} benzene]; m.p. $168-170^{\circ}C,\;C_{31}H_{22}O_8N_4Cl_6$ : 2,2'-Methylene-bis [3, 4, 6-trichloro ${{\beta}-(o-chloro-p-nitroanilino)$ propionoxy} benzene]; m.p. $170.5-172.5^{\circ}C,\;C_{31}H_{20}O_8N_4Cl_8$ : 2,2'-Methylene-bis [3, 4, 6-trichloro ${{\beta}-(c-methyl-p-nitroanilino)$ propionoxy} benzene]; m.p. $163-164^{\circ}C,\;C_{33}H_{26}O_8N_4Cl_6$-were synthesized by Mannich reaction from 2,2'-Methylene-bis (3, 4, 6-trichloroacetoxy benzene) and their antimicrobial activities against the microorganisms Staphylococcus aureus, Escherichia coli, Bacillus subtilis Natto, Brevibacterium ammoniagenes, Candida tropicalis, Rhodotorula glutinis, Pseudomonas ovalis, Aspergillus candidus Link, Aspergillus awamori Nakazawa. Aspergillus niger var. Tieghem, Aspergillus usami Sakakuchi, Penicillium notatum-were tested. 2,2'-methylene-bis [3, 4, 6-trichloro ${{\beta}-(o-nitroanilino)$ propionoxy} benzene] showed a strong antimicrobial activity against Bacilus subtilis Natto and Brevibacterium ammoniagenes.

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Effect of Incubation Period, Temperature and pH on Mycelial Growth of Cylindrocarpon destructans (Zinssm.) Scholten Causing Root-rot of Ginseng (배양기간, 온도, pH가 인삼 근부병균 Cylindrocarpon destructans (Zinssm.) Scholten의 균사생육에 미치는 영향)

  • 조대휘;안일평
    • Journal of Ginseng Research
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    • v.19 no.2
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    • pp.181-187
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    • 1995
  • Cylindvocarpon destmtalns isolate CY-92-01, pathogen of root-rot of Panax ginseng showed t the maximum mycelial growth on the Czapek solution agar among the thirteen kinds of media. Five isolates (Isolate CY-92-01, CY-92-03, CY-92-07, CY-94-01, CY-94-02) of C. destructan from various growth stages of p. ginseng recovered from several geographical sites also showed maximum growth in the Czapek-Dox broth compared with potato dextrose broth and V-8 juice broth. Rapid growth rate was maintained until 12 days after inoculation on the Czapek-Dox broth and mycelial weight was somewhat constant until 20 days. After 30 days of incubation, the mycelial weight began to decrease. The fungal growth occurred from 5$^{\circ}C$ to $25^{\circ}C$ and optimum temperature for growth was 2$0^{\circ}C$. Mycelial weight orderly decreased at 15, 25, 10, and 5$^{\circ}C$. Quantitative measurement was impossible at 5$^{\circ}C$. No fungal growth was occurred at the temperature higher than 3$0^{\circ}C$. Growth was observed at all tested pH ranges from 2.8 to 8.0. Optimum pH for growth was 4.0~5.0 followed by pH 3.3~3.5 and 5.4~6.0. The least growth occurred at pH 2.8.

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Analytical Model of Breakdown Voltages for 6H-SiC $p^{+}n$ Junction (6H-SiC $p^{+}n$ 접합의 항복 전압을 위한 해석적 모형)

  • Jeong, Yong-Seong
    • Journal of the Institute of Electronics Engineers of Korea SD
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    • v.38 no.6
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    • pp.398-403
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    • 2001
  • In this paper, effective ionization coefficient for 6H-SiC is determined. Analytical formulas for the parallel plane breakdown voltage of the 6H-SiC p+n junction are derived by employing the ionization coefficients. The analytical breakdown voltages show good agreement with the numerical results of Dmitriev's[3]and the experimental results of Cree Research[9]over the doping range from 10$^{15}$ cm$^{-3}$ to 10$^{18}$ cm$^{-3}$.

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