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ISOLATION AND IDENTIFICATION OF ANAEROBIC RUMEN BACTERIUM, ACTINOMYCES SP. 40 AND ENZYMATIC PROPERTIES OF β-1, 4-ENDOGLUCANASE

  • Min, H.K.;Choi, Y.J.;Ha, J.K.;Cho, K.K.;Kwon, Y.M.;Chang, Y.H.;Lee, S.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.7 no.3
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    • pp.373-382
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    • 1994
  • A bacterial strain No. 40, which produced extracellular endoglucanase, was isolated from the rumen of Korean native goals and identified to be a genus of Actinomyces sp. The optimum conditions for endoglucanase production in PY-CMC medium were initial pH of 7.0 and 4 days of cultivation at $39^{\circ}C$. When localization of endoglucanase activity of Actinomyces sp. was determined, 68% of the enzyme activity was found in the extracellular fraction, 11% of the activity was detected in the periplasmic space and the remaining activity was in the intracellular and cell-bound fractions. The maximal endoglucanase activity was observed at pH 5.0 and it was most s table at pH 5.0. The optimum temperature of this enzyme activity was $55^{\circ}C$, but enzyme activity was gradually lost at temperature above $60^{\circ}C$. The crude enzyme was activated by addition of 10 mM cysteine and 10 mM DTT. But it was inhibited by addition of 10 mM $Cu^{{+}{+}}$ and $Fe^{{+}{+}}$. This crude enzyme could digest carboxymethylcellulose (CMC), and degrade xylan, avicel, pNPG, and pNPC to a less extent.

Effect of Mercuric Chloride (In Vitro) on the Function of Human Polylnorphonuclear Leukocytes(PMNs) (수은이 시험관내 사람 다형핵백혈구의 기능에 미치는 영향)

  • 한형미;윤은이;김순한;김옥연;김효정;선우연
    • Biomolecules & Therapeutics
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    • v.1 no.2
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    • pp.131-136
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    • 1993
  • In the present study, the effect of $HgCl_2$on the function of human peripheral polymorphonuclear leukocytes(PMNs) was examined. PMNs were isolated from human peripheral blood with density centrifugation in Ficoll-Paque. The cells were then incubated with $0.5{\sim}5{\mu}M\;HgCl_2$and glass adherence, chemotactic activity and erythrocyte-antibody rosette forming activity were measured. $HgCl_2$ decreased the function of PMNs in all three aspects tested. $HgCl_2$significantly diminished glass adherence(40.5 {\mu}M: 92{\pm}12%$ (percentage of control, $mean{\pm}$ S.D.); 41 {\mu}M: 46{\pm}11%,$ P<0.01; $3{\mu}M: 35{\pm}7%,$P<0.01;$5{\mu}M:49{\pm}10%,$ P<0.01). Similarly, significant differences were observed in chemotactic activity after $HgCl_2$treatment compared with control (control: $0.95{\pm}0.14mm; 0.5 {\mu}M: 0.91{\pm}0.11 mm; 1 {\mu}M: 0.77{\pm}0.16mm, P<0.05; 3{\mu}M: 0.61{\pm} 0.06mm, P<0.01; 5{\mu}M: 0.15{\pm}0.03 mm, P<0.01).$ Also, 4HgCl_2$decreased the percentage of rosette-forming PMNs, indicating diminished phagocytic activity of PMNs upon $HgCl_2$ exposure compared with control (control: $58{\pm}4%; 1{\mu}M: 53{\pm}4%, p<0.05; 3{\mu}M: 49{\pm}3%, P<0.01; 5{\mu}M: 46{\pm}3%, P<0.01).$ Cell viability was not antered after $HgCl_2$treatment (483{\pm}5%$ viability in control PMNs versus $81{\pm}8%$ viability in $5{\mu}M$ Hg-treated PMNs), suggesting that the impaired PMN function after $HgCl_2$treatment was not due to nonspecific cytotoxicity induced by $HgCl_2$. $HgCl_2$-induced decrease in the function of PMNs may have some implications in depressed host susceptibilityupon bacterial challenge after mercury exposure.

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Biochemical Characterization of an Extracellular ${\beta}$-Glucosidase from the Fungus, Penicillium italicum, Isolated from Rotten Citrus Peel

  • Park, Ah-Reum;Hong, Joo-Hee;Kim, Jae-Jin;Yoon, Jeong-Jun
    • Mycobiology
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    • v.40 no.3
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    • pp.173-180
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    • 2012
  • A ${\beta}$-glucosidase from Penicillium italicum was purified with a specific activity of 61.8 U/mg, using a chromatography system. The native form of the enzyme was an 88.5-kDa tetramer with a molecular mass of 354 kDa. Optimum activity was observed at pH 4.5 and $60^{\circ}C$, and the half-lives were 1,737, 330, 34, and 1 hr at 50, 55, 60, and $65^{\circ}C$, respectively. Its activity was inhibited by 47% by 5 mM $Ni^{2+}$. The enzyme exhibited hydrolytic activity for p-nitrophenyl-${\beta}$-D-glucopyranoside (pNP-Glu), p-nitrophenyl-${\beta}$-D-cellobioside, p-nitrophenyl-${\beta}$-D-xyloside, and cellobiose, however, no activity was observed for p-nitrophenyl-${\beta}$-D-lactopyranoside, p-nitrophenyl-${\beta}$-D-galactopyranoside, carboxymetyl cellulose, xylan, and cellulose, indicating that the enzyme was a ${\beta}$-glucosidase. The $k_{cat}/K_m\;(s^{-1}mM^{-1})$ values for pNP-Glu and cellobiose were 15,770.4 mM and 6,361.4 mM, respectively. These values were the highest reported for ${\beta}$-glucosidases. Non-competitive inhibition of the enzyme by both glucose ($K_i=8.9mM$) and glucono-${\delta}$-lactone ($K_i=11.3mM$) was observed when pNP-Glu was used as the substrate. This is the first report of non-competitive inhibition of ${\beta}$-glucosidase by glucose and glucono-${\delta}$-lactone.

Ginsenoside Rg1 Induces Autophagy in Colorectal Cancer through Inhibition of the Akt/mTOR/p70S6K Pathway

  • Ruiqi Liu;Bin Zhang;Shuting Zou;Li Cui;Lin, Lin;Lingchang Li
    • Journal of Microbiology and Biotechnology
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    • v.34 no.4
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    • pp.774-782
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    • 2024
  • This study aimed to elucidate the anti-colon cancer mechanism of ginsenoside Rg1 in vitro and in vivo. Cell viability rate was detected using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) tetrazolium assay. The inhibitory effect of ginsenoside Rg1 against CT26 cell proliferation gradually increased with increasing concentration. The in vivo experiments also demonstrated an antitumor effect. The monodansylcadaverine (MDC), transmission electron microscopy (TEM), and expression of autophagy marker proteins confirmed that ginsenoside Rg1 induced autophagy in vitro. Ginsenoside Rg1 induced autophagy death of CT26 cells, but this effect could be diminished by autophagy inhibitor (3-methyladenine, 3-MA). Additionally, in a xenograft model, immunohistochemical analysis of tumor tissues showed that the LC3 and Beclin-1 proteins were highly expressed in the tumors from the ginsenoside Rg1-treated nude mice, confirming that ginsenoside Rg1 also induced autophagy in vivo. Furthermoer, both in vivo and in vitro, the protein expressions of p-Akt, p-mTOR, and p-p70S6K were inhibited by ginsenoside Rg1, which was verified by Akt inhibitors. These results indicated that the mechanism of ginsenoside Rg1 against colon cancer was associated with autophagy through inhibition of the Akt/mTOR/p70S6K signaling pathway.

The Change of Serum Soluble E-selectin in Kawasaki Disease (가와사끼병에서의 혈청 Soluble E-selectin의 변화)

  • Jeong, Jae-Ho;Cho, Eun-Young;Lim, Jae-Woo;Cheon, Eun-Jeong;Ko, Kyong-Og;Lee, Kyung-Il
    • Clinical and Experimental Pediatrics
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    • v.48 no.5
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    • pp.539-544
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    • 2005
  • Purpose : The aim of this study was to investigate the pathophysiologic role of serum E-selectin, vascular endothelial growth factor(VEGF)-induced cell adhesion mollecule in Kawasaki disease(KD) and to look for the evidence of direct relationship between the plasma levels of soluble E-selectin and the incidence of coronary artery lesion(CAL). Methods : Changes in plasma levels of sE-selectin(n=98) over time were measured by enzyme-linked immunosorbent assay(ELISA) in 23 patients with acute KD and 25 age-matched febrile children. Results : Compared with control values, the peak levels of plasma sE-selectin were significantly elevated($mean{\pm}S.E$. : $22.89{\pm}12.53ng/mL$ vs $10.65{\pm}3.42ng/mL$, P=0.01) in KD. 5 patients with CAL, plasma sE-selectin levels before treatment were higher than in 18 patients without CAL($mean{\pm}S.E$. : $39.43{\pm}15.08ng/mL$ and $19.00{\pm}8.32ng/mL$, respectively; P=0.01). Plasma sE-selectin declined rapidly in the majority of KD patients regardless of the presence of CAL. Plasma sE-selectin levels after treatment and convalesent period were similar in KD patients with and without CAL. The plasma levels sE-selectin were correlated with those of white blood cell count(r=0.299, P<0.05), CRP(r=0.430, P<0.05), serum albumin(r=-0.483, P<0.05), serum protein(r=-0.502, P<0.05) and hemoglobin(r=-0.372, P<0.05) not with those of ESR, platelet, or duration of fever. There were significant differences in the initial level of serum sE-selectin between KD with and without CAL($mean{\pm}S.E$. : $39.44{\pm}15.08ng/mL$ vs. $19.00{\pm}17.18ng/mL$) in multivariated linear tests. Conclusion : Plasma sE-selectin levels were significantly higher in KD than in other febrile illness. Higher plamsa levels of sE-selectin may have potential as a predictor of CAL in patients with KD.

The Effect of Single and Mixed Microbial Inoculation on the in situ Fiber Digestibility and Silage of Rice Straw Contaminated Mycotoxins (단일 및 복합 미생물 접종이 곰팡이독소 오염 볏짚의 사일리지 및 In situ 섬유소 소화율에 미치는 영향)

  • Ha Guyn Sung
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.42 no.4
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    • pp.229-236
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    • 2022
  • This study was conducted to evaluate the efficacy of adding the microbial inoculants to silage for reducing mycotoxins in rice straw silage. When a single agent of L. plantarum and a mixed agent of L. plantarum and S. cerevisiae were added in rice straw silage contaminated mycotoxins, it had an effect on silage fermentation and fiber degradation as well as mycotoxin reduction. Among the mycotoxins, only ochratoxin A and zearalenone were found in the test sample. Ochratoxin A and zearalenone showed a decreasing trend with the addition of silage inoculants compared to the control groups (38.11±2.22 and 633.67±50.30 ㎍/kg), and there was a significant difference at the mixed agents; 27.78±2.28 and 392.72±25.04 ㎍/kg, respectively (p<0.05). The pH was lower in the single agent and the mixed agent compared to the control (p<0.05). The concentration of lactic acid was higher in the single agent (11.73±0.31 mM) than in the control group (8.18±0.93 mM), and the highest concentration was 16.01±0.88 mM in the mixed agent (p<0.05). Acetic acid and propionic acid were found to be significantly lowered with the addition of silage inoculants (p<0.05). Total VFA was also lower at the addition of silage inoculants than the control group (p<0.05). The rumen in situ dry matter degradation of NDF and ADF was maintained at the highest levels of the mixed agent during the culture period, followed by the single agent and the control group at the lowest level. NDF and ADF degradation showed a significant difference at all time points after 12 and 24 hours of culture, respectively (p<0.05). The study results showed that the silage inoculants had the positive effects on quality increasing of rice straw silage; fermentative charateristics, fiber degradation and mycotoxins reduction. Ochratoxin A and zearalenone were greater reduction by adding bacterial inoculants of silage. Therefore it is considered that L. plantarum and S. cerevisiae will improve the quality and stability with remediation of mycotoxin in silage.

Identification of Cultivate Sites for Job's-tears (Coix lachrymajobi var. mayuen) using Capillary Electrophoresis (Capillary electrophoresis를 이용한 율무의 원산지 판별)

  • Rhyu, Mee-Ra;Kim, Eun-Young;Kim, Sang-Sook
    • Korean Journal of Food Science and Technology
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    • v.34 no.5
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    • pp.787-791
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    • 2002
  • Optimal extraction, separation, and capillary rinsing conditions for capillary electrophoresis (CE) were established to identify the cultivation site (domestic vs. foreign) of Job's-tears (Coix lachrymajobi var. mayuen) using 240 samples (domestic sample n = 121, foreign sample n = 119). Job's-tears was extracted with 30% ethanol and separated on a $50-{\mu}m-I.D.$ untreated fused-silica capillary. Optimal analytic conditions were: temperature, $45^{\circ}C$; voltage, 15 kV; detector rise time, 0.1 sec; and pressure injection, 20 sec. Separation of peak investigated using 0.1 M phosphate buffer (pH 2.5) containing 0.05% hydroxypropylmethylcellulose (P buffer) revealed the optimal separation buffer was P buffer containing 26 mM hexane sulfonic acid with 30% methanol. Under the optimal conditions established for CE, the average correct identification percentage of domestic or foreign Job's-tears was 82%.

Analysis of Weathered State on a Halo Stone Buddha, Unju Temple of Hwasun, Korea Using Low Frequency Flaw Detector (저주파 결함 탐지기를 활용한 화순 운주사 광배석불의 풍화상태 분석)

  • Kang, Seong-Seung;Ko, Chin-Surk;Kim, Cheong-Bin;Jang, Bo-An
    • The Journal of Engineering Geology
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    • v.23 no.3
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    • pp.235-246
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    • 2013
  • P-wave velocity was measured by the low frequency flaw detector in order to analyze the weathered state of a halo stone Buddha, Unju temple, Hwasun, Korea. By the results of laboratory tests on a fresh acidic tuff with the same rock of a halo stone Buddha, average absorption, average P-wave velocity, and average uniaxial compressive strength were 5.38%, 4,008 m/s, and 70.1 MPa, respectively. The results correspond to moderately strong rock. Average P-wave velocity of a halo stone Buddha measured by the low frequency flaw detector was 2,257 m/s in the left zone, 3,437 m/s in the right zone, and 2,802 m/s overall. Weathering index of a halo stone Buddha was 0.45 in the left zone, 0.21 in the right zone, and 0.33 overall. Comparing the results of a halo stone Buddha with them of laboratory tests, weathered state of a halo stone Buddha was analyzed highly weathered state in the left zone and moderately weathered state in the right zone. Furthermore, it suggests that the left zone of a halo stone Buddha was affected weathering more than the right one. Overall a halo stone Buddha corresponds to moderately weathered state of weathering degrees. In conclusion, it is considered that low frequency flaw detector may be applicable as a valuable method on analyzing the P-wave velocity of the stone cultural heritage with an irregular surface.

A SOLUTION OF EGGERT'S CONJECTURE IN SPECIAL CASES

  • KIM, SEGYEONG;PARK, JONG-YOULL
    • Honam Mathematical Journal
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    • v.27 no.3
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    • pp.399-404
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    • 2005
  • Let M be a finite commutative nilpotent algebra over a perfect field k of prime characteristic p and let $M^p$ be the sub-algebra of M generated by $x^p$, $x{\in}M$. Eggert[3] conjectures that $dim_kM{\geq}pdim_kM^p$. In this paper, we show that the conjecture holds for $M=R^+/I$, where $R=k[X_1,\;X_2,\;{\cdots},\;X_t]$ is a polynomial ring with indeterminates $X_1,\;X_2,\;{\cdots},\;X_t$ over k and $R^+$ is the maximal ideal of R generated by $X_1,\;X_2,{\cdots},\;X_t$ and I is a monomial ideal of R containing $X_1^{n_1+1},\;X_2^{n_2+1},\;{\cdots},\;X_t^{n_t+1}$ ($n_i{\geq}0$ for all i).

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$C^*$-ALGEBRAS ASSOCIATED WITH LENS SPACES

  • Boo, Deok-Hoon;Oh, Sei-Qwon;Park, Chun-Gil
    • Communications of the Korean Mathematical Society
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    • v.13 no.4
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    • pp.759-764
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    • 1998
  • We define the rational lens algebra (equation omitted)(n) as the crossed product by an action of Z on C( $S^{2n+l}$). Assume the fibres are $M_{ k}$/(C). We prove that (equation omitted)(n) $M_{p}$ (C) is not isomorphic to C(Prim((equation omitted)(n))) $M_{kp}$ /(C) if k > 1, and that (equation omitted)(n) $M_{p{\infty}}$ is isomorphic to C(Prim((equation omitted)(n))) $M_{k}$ /(C) $M_{p{\infty}}$ if and only if the set of prime factors of k is a subset of the set of prime factors of p. It is moreover shown that if k > 1 then (equation omitted)(n) is not stably isomorphic to C(Prim(equation omitted)(n))) $M_{k}$ (c).

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