• Title/Summary/Keyword: GS1

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THE COMPARISON OF STREPTOCOCCUS MUTANS ISOLATED FROM OCCLUSAL SURFACES OF CARIES AND NON-CARIES TEETH (우식치아와 정상치아의 교합면에서 분리한 Streptococcus mutans의 비교)

  • Park, Ho-Won;Jung, Tae-Sung;Jung, Jin;Kim, Shin
    • Journal of the korean academy of Pediatric Dentistry
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    • v.28 no.1
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    • pp.129-141
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    • 2001
  • When oral microorganisms were sampled from occlusal surfaces of caries and non-caries teeth, $3.43\times10^5$ CFU and $3.47\times10^3$ CFU of bacteria were counted on MSB agar plates, respectively. All the 20 colonies isolated from a caries surface were Streptococcus mutans but, only two of 20 colonies were identified as Streptococcus mutans by API test. S. mutans SM1 from caries tooth and S. mutans SM2 from non-caries tooth showed the same results except for $\alpha-galactosidase$ activity on sugar fermentation tests and biochemical tests. For the bacterial replication, both SM1 and SM2 were actively multiplicated at pH 5.5. And the viability of SM1 was high at 20% of sucrose, while that of SM2 was high at 5% of sucrose in the media. SM1 actively replicated at 16mM of $CaCl_2$, 160mM of KCl, and 6.4mM of $MgCl_2$, and the replication of SM2 was increased at 16mM of $CaCl_2$, 40mM of KCl, 6.4mM of $MgCl_2$. At 1mM of sodium bicarbonate and sodium phosphate, both bacteria were actively multiplicated. SM1 and SM2 were actively replicated at 1mM and 10mM of Tris, respectively. For potassium phosphate buffer, SM1 grew well proportionally to the concentration up to 100mM, while the growth of SM2 were inhibited by the increase of concentration. The 4.6 kb of gtf gene was amplified with a pair of primer, gtfB-F961 and gtfC-R5574 by polymerase chain reaction from the chromosomal DNA of SM1 and SM2. When 4.6kb bands were eluted from gel and were treated with restriction enzyme, EcoR I produced the same RFLP like 0.8kb and 3.8kb of DNA fragments for S. mutans GS-5, SM1 and SM2. By Hind III, the PCR products weren't digested for S. mutans GS-5 and SM1, but 3 fragments such as 2.4kb, 1.8kb and 400bp were examined for SM2. These results indicated the difference between gtf genes of SM1 and SM2. BamH I treatment showed 4 fragments for SM1 and SM2, while the 3 fragments for S. mutans GS-5. The PCR products were not digested by Kpn I, Sma I, Xho I and Pst I.

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Development of Marker-free TaGlu-Ax1 Transgenic Rice Harboring a Wheat High-molecular-weight Glutenin Subunit (HMW-GS) Protein (벼에서 밀 고분자 글루테닌 단백질(TaGlu-Ax1) 발현을 통하여 쌀가루 가공적성 증진을 위한 마커프리(marker-free) 형질전환 벼의 개발)

  • Jeong, Namhee;Jeon, Seung-Ho;Kim, Dool-Yi;Lee, Choonseok;Ok, Hyun-Choong;Park, Ki-Do;Hong, Ha-Cheol;Lee, Seung-Sik;Moon, Jung-Kyung;Park, Soo-Kwon
    • Journal of Life Science
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    • v.26 no.10
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    • pp.1121-1129
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    • 2016
  • High-molecular-weight glutenin subunits (HMW-GSs) are extremely important determinants of the functional properties of wheat dough. Transgenic rice plants containing a wheat TaGlu-Ax1 gene encoding a HMG-GS were produced from the Korean wheat cultivar ‘Jokyeong’ and used to enhance the bread-making quality of rice dough using the Agrobacterium-mediated co-transformation method. Two expression cassettes with separate DNA fragments containing only TaGlu-Ax1 and hygromycin phosphotransferase II (HPTII) resistance genes were introduced separately into the Agrobacterium tumefaciens EHA105 strain for co-infection. Rice calli were infected with each EHA105 strain harboring TaGlu-Ax1 or HPTII at a 3:1 ratio of TaGlu-Ax1 and HPTII. Among 210 hygromycin-resistant T0 plants, 20 transgenic lines harboring both the TaGlu-Ax1 and HPTII genes in the rice genome were obtained. The integration of the TaGlu-Ax1 gene into the rice genome was reconfirmed by Southern blot analysis. The transcripts and proteins of the wheat TaGlu-Ax1 were stably expressed in rice T1 seeds. Finally, the marker-free plants harboring only the TaGlu-Ax1 gene were successfully screened in the T1 generation. There were no morphological differences between the wild-type and marker-free transgenic plants. The quality of only one HMW-GS (TaGlu-Ax1) was unsuitable for bread making using transgenic rice dough. Greater numbers and combinations of HMW and LMW-GSs and gliadins of wheat are required to further improve the processing qualities of rice dough. TaGlu-Ax1 marker-free transgenic plants could provide good materials to make transgenic rice with improved bread-making qualities.

Influences of Culture Medium Components on the Production Poly (γ-Glutamic Acid) by Bacillus subtilis GS-2 Isolated Chungkookjang (청국장에서 분리한 Bacillus subtilis GS-2에 의한 Poly(γ-Glutamic Acid) 생산의 최적 배양조건)

  • Bang, Byung-Ho;Rhee, Moon-Soo;Kim, Kwan-Pil;Yi, Dong-Heui
    • The Korean Journal of Food And Nutrition
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    • v.25 no.3
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    • pp.677-684
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    • 2012
  • A bacterium strain GS-2 isolated from the Korean traditional seasoning food, Chungkookjang and was determined to produce large amounts of ${\gamma}$-PGA with high productivity when provided with simple nutrients (L-glutamic acid 2.0%, glucose 1.0%, $NH_4Cl$ 0.5%, $KH_2PO_4$ 0.05%, $MgSO_4{\cdot}7H_2O$ 0.01%, pH 7.0). In this study, the culture medium for this strain was optimized for the production of ${\gamma}$-PGA. The Bacillus subtilis GS-2 required supplementation with L-glutamic acid and other nutrients for maximal production of ${\gamma}$-PGA. The optimal culture conditions for ${\gamma}$-PGA production were a 48 hr culture time, a temperature of $33^{\circ}C$ and initial pH of 6.5 by rotary shaking (220 rpm). A maximum ${\gamma}$-PGA production of 31.0 $g/{\ell}$ was obtained with L-glutamic acid (30 $g/{\ell}$), sucrose (the main carbon source, 30 $g/{\ell}$), $NH_4Cl$ (the main nitrogen source, 2.5 $g/{\ell}$), $KH_2PO_4$ (1.5 $g/{\ell}$) and $MgSO_4{\cdot}7H_2O$ (0.15 $g/{\ell}$) in the culture medium.

ANTIBACTERIAL EFFECT OF POLYPHOSPHATES ON MUTANS STREPTOCOCCI (Mutans streptococci에 대한 polyphosphate의 항균효과)

  • Kang, Kye-Sook;Choi, Yeong-Chul
    • Journal of the korean academy of Pediatric Dentistry
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    • v.30 no.1
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    • pp.80-91
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    • 2003
  • Mutans streptococci, especially S. mutans and S. sobrinus strongly implicated in pathogenesis of dental caries, the major cause of tooth loss in children. Use of an antibacterial agent controlling dental caries has been rationalized. The present study was performed to observe the antibacterial effect of inorganic polyphosphates (polyP) on S. mutans and S. sobrinus. S. mutans GS5 and S. sobrinus 6715 were grown in brain-heart infusion broth with or without polyP. Minimal inhibitory concentration (MIC) of polyP for S. mutans GS5 was determined to be 0.08% and that for S. sobrius 6715 was 0.17%. PolyP 15 added to the growing culture of S. mutans GS5 and S. sobrinus 6715 at their exponential phase was as effective in inhibiting the growth of S. mutans GS5 and S. sobrinus 6715 as polyP added at the very beginning of the culture. More than 85% of the cells lost their viability determined by viable cell count when polyP 15 was added to the culture of growing S. mutans GS5 at MIC, suggesting that polyP 15 has bacterial effect on the bacterium. And more than 99.9% of the cells lost their viability determined by viable cell count when polyP 15 was added to the culture of growing S. sobrinus 6715 at MIC, suggesting that polyP 15 has bacterial effect on the bacterium. Intracellular nucleotide release from S. mutans CS5 and S. sobrinus 6715 was increased in the presence of polyP 15 for 5h but was not really reversed by the addition of divalent cations like $Ca^{++}\;and\;Mg^{++}$. The majority of the cells appeared to be atypical in their shape, demonstrating accumulation of highly electron-dense granules and ghost cells. The overall results suggest that polyP have a strong bactericidal activity against S. mutans and S. sobrinus in which lysis in relation to chelation may not play the major role but unknown mechanism that possibly affects the viability of the bacterium may be involved. PolyP may be used as an agent for prevention of dental caries.

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Analysis of Calcite Twins as Indicators of Paleostress History (고응력의 지시자로서 방해석 쌍정 분석연구)

  • Park, Young-Seog;Jang, Bo-An;Kim, Cheong-Bin;Kang, Seong-Seung
    • Economic and Environmental Geology
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    • v.40 no.4
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    • pp.461-471
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    • 2007
  • A temperature of deformation and the state and direction of paleostress at that time when twins in calcite grains had been produced were observed, using analysis of calcite twins as indicators of paleostress history. The study was performed with the target of carbonate rocks distributed randomly small size in the southern area of south Korea. Considering the appearance of twins (thin or thick straight twins with one or two twin sets), average twin strain (1.235-7.453%), thickness ($0.77-1.94{\mu}m$) and intensity (25.26-41.99 twins/mm) from the results of calculated calcite twins, it is estimated that calcite twins were produced under temperatures lower than approximately $150-200^{\circ}C$. In the magnitudes and directions of principal strains, the maximum shortening strain axis ($e_3\leftrightarrow{\sigma}_1$) is approximately N-S direction in the GS-1 area in the southern Gyeongsang Basin as well as in the BS-1 area in the southern Yongnam Massif, whereas E-W direction in the NR-1 area in the southwestern Ogcheon Fold Belt. In case of the maximum extension strain axis ($e_1\leftrightarrow{\sigma}_3$), it is oriented in NW-SE and NE-SW directions in the GS-1 and BS-1 area, respectively, and in N-S direction in the NR-1 area. That is, it is suggested that the paleostress which produced the calcite twins may be applied at least more than two times in the study area.

Effect of Cytokines on the Growth and Differentiation of the Glial Cells from Rat Brain in Culture (랫트 배양 신경교세포의 성장 및 분화에 대한 Cytokine의 효과)

  • Kim, Hae-Kyoung;Youn, Yong-Ha;Kang, Shin-Chung;Park, Chan-Woong;Kim, Yong-Sik
    • The Korean Journal of Pharmacology
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    • v.32 no.2
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    • pp.177-188
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    • 1996
  • The effects of cytokines on the growth and differentiation of glial cells in culture were evaluated to confirm that cytokines could modify the number and function of glial cells. Proliferation of glial cells was determined by the $^3H-thymidine$ uptake and the double immunostain with anti-cell specific marker and anti-bromodeoxyuridine(BrdU) antibody. To check the effect on the differentiation of glial cells, the amount of glial fibrillar acidic protein(GFAP) and the activity of glutamine synthetase(GS) were measured in astrocytes. And also the amounts of myelin basic protein(MBP) and the activity of 2',3'-cyclic nucleotide phosphohydrolase(CNPase) were measured in oligodendrocytes. Among the cytokines used, only interleukin-$1{\beta}(IL-1{\beta})$ stimulated the growth of type 1 and type 2 astrocyte as well as 0-2A precursor cell. When the functional changes in these glial cells by cytokines were tested, $IL-1{\beta}$ did not increase GFAP content in type 1 and type 2 astrocyte, but $IL-1{\beta}$ increased GS activity in type 1 astrocyte, and slightly decreased this enzyme activity in type 2 astrocyte. Also interleukin-2(IL-2) and $interferon-{\gamma}$ $(IFN-{\gamma})$ inhibited the activity of GS in type 1 and type 2 astrocyte. On the other hand, all cytokines used did not modify the growth and differentiation in oligodendrocytes. From these results we could suggest that $IL-1{\beta}$ increases the growth of type 1 and type 2 astrocyte and also promotes the development for 0-2A precursor cell to type 2 astrocyte.

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Development of Marker-free Transgenic Rice for Increasing Bread-making Quality using Wheat High Molecular Weight Glutenin Subunits (HMW-GS) Gene (밀 고분자 글루테닌 유전자를 이용하여 빵 가공적성 증진을 위한 마커 프리 형질전환 벼의 개발)

  • Park, Soo-Kwon;Shin, DongJin;Hwang, Woon-Ha;Oh, Se-Yun;Cho, Jun-Hyun;Han, Sang-Ik;Nam, Min-Hee;Park, Dong-Soo
    • Journal of Life Science
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    • v.23 no.11
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    • pp.1317-1324
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    • 2013
  • High-molecular weight glutenin subunits (HMW-GS) have been shown to play a crucial role in determining the processing properties of the wheat grain. We have produced marker-free transgenic rice plants containing a wheat Glu-1Bx7 gene encoding the HMG-GS from the Korean wheat cultivar 'Jokyeong' using the Agrobacterium-mediated co-transformation method. The Glu-1Bx7-own promoter was inserted into a binary vector for seed-specific expression of the Glu-1Bx7 gene. Two expression cassettes comprised of separate DNA fragments containing only Glu-1Bx7 and hygromycin phosphotransferase II (HPTII) resistance genes were introduced separately to the Agrobacterium tumefaciens EHA105 strain for co-infection. Each EHA105 strain harboring Glu-1Bx7 or HPTII was infected to rice calli at a 3:1 ratio of Glu-1Bx7 and HPTII, respectively. Then, among 216 hygromycin-resistant $T_0$ plants, we obtained 24 transgenic lines with both Glu-1Bx7 and HPTII genes inserted into the rice genome. We reconfirmed integration of the Glu-1Bx7 gene into the rice genome by Southern blot analysis. Transcripts and proteins of the wheat Glu-1Bx7 were stably expressed in the rice $T_1$ seeds. Finally, the marker-free plants harboring only the Glu-1Bx7 gene were successfully screened at the $T_1$ generation.

Major ginsenosides from Panax ginseng promote aerobic cellular respiration and SIRT1-mediated mitochondrial biosynthesis in cardiomyocytes and neurons

  • Huang, Qingxia;Lou, Tingting;Lu, Jing;Wang, Manying;Chen, Xuenan;Xue, Linyuan;Tang, Xiaolei;Qi, Wenxiu;Zhang, Zepeng;Su, Hang;Jin, Wenqi;Jing, Chenxu;Zhao, Daqing;Sun, Liwei;Li, Xiangyan
    • Journal of Ginseng Research
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    • v.46 no.6
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    • pp.759-770
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    • 2022
  • Background: Aerobic cellular respiration provides chemical energy, adenosine triphosphate (ATP), to maintain multiple cellular functions. Sirtuin 1 (SIRT1) can deacetylate peroxisome proliferator-activated receptor gamma coactivator 1 alpha (PGC-1α) to promote mitochondrial biosynthesis. Targeting energy metabolism is a potential strategy for the prevention and treatment of various diseases, such as cardiac and neurological disorders. Ginsenosides, one of the major bioactive constituents of Panax ginseng, have been extensively used due to their diverse beneficial effects on healthy subjects and patients with different diseases. However, the underlying molecular mechanisms of total ginsenosides (GS) on energy metabolism remain unclear. Methods: In this study, oxygen consumption rate, ATP production, mitochondrial biosynthesis, glucose metabolism, and SIRT1-PGC-1α pathways in untreated and GS-treated different cells, fly, and mouse models were investigated. Results: GS pretreatment enhanced mitochondrial respiration capacity and ATP production in aerobic respiration-dominated cardiomyocytes and neurons, and promoted tricarboxylic acid metabolism in cardiomyocytes. Moreover, GS clearly enhanced NAD+-dependent SIRT1 activation to increase mitochondrial biosynthesis in cardiomyocytes and neurons, which was completely abrogated by nicotinamide. Importantly, ginsenoside monomers, such as Rg1, Re, Rf, Rb1, Rc, Rh1, Rb2, and Rb3, were found to activate SIRT1 and promote energy metabolism. Conclusion: This study may provide new insights into the extensive application of ginseng for cardiac and neurological protection in healthy subjects and patients.