• Title/Summary/Keyword: -galactosidase activity

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Influence of 10-Methacryloyloxydecyl Dihydrogen Phosphate on Cellular Senescence in Osteoblast-Like Cells

  • Ju Yeon Ban;Sang-Im Lee
    • Journal of dental hygiene science
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    • v.23 no.4
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    • pp.264-270
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    • 2023
  • Background: Resin-based dental materials release residual monomers or other substances from incomplete polymerization into the oral cavity, thereby causing adverse biological effects on oral tissue. 10-Methacryloyloxydecyl dihydrogen phosphate (10-MDP), an acidic monomer containing dihydrogen phosphate and methacrylate groups, is the most commonly used component of resin-based dental materials, such as restorative composite resins, dentin adhesives, and resin cements. Although previous studies have reported the cytotoxicity and biocompatibility in various cultured cells, the effects of resin monomers on cellular aging have not been reported to date. Therefore, this study aimed to investigate the effects of the resin monomer 10-MDP on cellular senescence and inflamm-aging in vitro. Methods: After stimulation with 10-MDP, MC3T3-E1 osteoblast-like cells were examined for cell viability by WST-8 assay and reactive oxygen species (ROS) production by flow cytometry. The protein and mRNA levels of molecular markers of aging were determined by western blotting and RT-PCR analysis, respectively. Results: Treatment with 0.05 to 1 mM 10-MDP for 24 hours reduced the survival of MC3T3-E1 cells in a concentration-dependent manner. The intracellular ROS levels in the 10-MDP-treated experimental group were significantly higher than those in the control group. 10-MDP at a concentration of 0.1 mM increased p53, p16, and p21 protein levels. Additionally, an aging pattern was observed with blue staining due to intracellular senescence-associated beta-galactosidase activity. Treatment with 10-MDP increased the levels of tumor necrosis factor-α, interleukin (IL)-1β, IL-6 and IL-8, however their expression was decreased by mitogen-activated-protein-kinase (MAPK) inhibitors. Conclusion: Taken together, these results suggest that the exposure of osteoblast-like cells to the dental resin monomer 10-MDP, increases the level of cellular senescence and the inflammatory response is mediated by the MAPK pathway.

Development of Cosmetic Ingredient by Fermented Paprika Juice (파프리카 발효즙의 화장품 소재개발 연구)

  • Bae, Soo Jung;Song, Min Hyeon;Oh, Jung Young;Bae, Jun Tae;Kim, Jin Hwa;Lee, Geun Soo
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.44 no.2
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    • pp.117-124
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    • 2018
  • In this study, cosmetic materials were developed using a new method of making juice through the fermentation of raw natural materials with microorganisms in order to supplement the advantages and disadvantages of an organic solvent extraction method and a microbial fermentation method. The natural products were selected from two colors (red, green) of paprika known to be rich in various colors and vitamins. The microorganisms used for fermentation were fermented by inoculating paprika with lactic acid bacteria (Lactobacillus plantarum) having sugar-hydrolyzed ability. First, we investigated the changes of physiologically active substances of two kinds of paprika juice and two kinds of fermented paprika juice. Total phenols content and total flavonoids content were higher in the fermented paprika juice than in the paprika juice, and especially in the fermented red paprika juice. Free radical scavenging effect and lipid peroxidation inhibitory effect were also showed an excellent antioxidative effect on paprika fermented juice, among which the effect of red paprika fermentation juice was the highest. The expression of MMP-1 in fermented red paprika juice with high antioxidant activity was inhibited by concentration-dependent expression of MMP-1 mRNA and MMP-1 protein. In the glycation experiments with aging, the anti-glycation effect of fermented paprika juice was highly inhibited by the production of advanced glycation end-products (AGEs), which was closely related to the antioxidant effect. In addition, the activity of senescence-associated ${\beta}$-galactosidase (SA-${\beta}$-gal), an indicator of cell senescence, was measured using human dermal fibroblast (HDF). The results showed that the cell senescence was inhibited when the cells were treated with fermented paprika juice. In conclusion, fermented paprika juice using lactic acid bacteria showed better antioxidative and anti-aging effects than paprika juice. Among them, fermented red paprika juice has the best antioxidant and anti-aging effect and can be applied as natural new material of antioxidant and anti-aging.

Korean Red ginseng prevents endothelial senescence by downregulating the HO-1/NF-κB/miRNA-155-5p/eNOS pathway

  • Kim, Tae-Hoon;Kim, Ji-Yoon;Bae, Jieun;Kim, Young-Mi;Won, Moo-Ho;Ha, Kwon-Soo;Kwon, Young-Guen;Kim, Young-Myeong
    • Journal of Ginseng Research
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    • v.45 no.2
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    • pp.344-353
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    • 2021
  • Background: Korean Red ginseng extract (KRGE) has beneficial effects on the cardiovascular system by improving endothelial cell function. However, its pharmacological effect on endothelial cell senescence has not been clearly elucidated. Therefore, we examined the effect and molecular mechanism of KRGE on the senescence of human umbilical vein endothelial cells (HUVECs). Methods: HUVECs were grown in normal or KRGE-supplemented medium. Furthermore, they were transfected with heme oxygenase-1 (HO-1) gene or treated with its inhibitor, a NF-κB inhibitor, and a miR-155-5p mimic or inhibitor. Senescence-associated characteristics of endothelial cells were determined by biochemical and immunohistochemical analyses. Results: Treatment of HUVECs with KRGE resulted in delayed onset and progression of senescence-associated characteristics, such as increased lysosomal acidic β-galactosidase and decreased telomerase activity, angiogenic dysfunction, and abnormal cell morphology. KRGE preserved the levels of anti-senescent factors, such as eNOS-derived NO, MnSOD, and cyclins D and A: however, it decreased the levels of senescence-promoting factors, such as ROS, activated NF-κB, endothelial cell inflammation, and p21 expression. The beneficial effects of KRGE were due to the induction of HO-1 and the inhibition of NF-κB-dependent biogenesis of miR-155-5p that led to the downregulation of eNOS. Moreover, treatment with inhibitors of HO-1, NF-κB, and miR-155-5p abolished the anti-senescence effects of KRGE. Conclusion: KRGE delayed or prevented HUVEC senescence through a signaling cascade involving the induction of HO-1, the inhibition of NF-κB-dependent miR-155-5p biogenesis, and the maintenance of the eNOS/NO axis activity, suggesting that it may protect against vascular diseases associated with endothelial senescence.

Comparative Analysis on Anti-aging, Anti-adipogenesis, and Anti-tumor Effects of Green Tea Polyphenol Epigallocatechin-3-gallate (녹차의 폴리페놀류인 에피갈로카테킨-3-갈레이트에 의한 항노화, 항비만 및 항암효과에 대한 비교 분석)

  • Lim, Eun-Ji;Kim, Min-Jae;Kim, Hyeon-Ji;Lee, Sung-Ho;Jeon, Byeong-Gyun
    • Journal of Life Science
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    • v.28 no.10
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    • pp.1201-1211
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    • 2018
  • The study compared the anti-aging, anti-adipogenesis, and anti-tumor effects of epigallocatechin-3- gallate (EGCG) in various cancer cell lines (SNU-601, MKN74, AGS, MCF-7, U87-MG, and A-549) and normal cell lines (MRC-5 fibroblasts, dental tissue-derived mesenchymal stem cells [DSC], and 3T3-L1 pro-adipocytes). Half inhibitory concentration ($IC_{50}$) values were significantly (p<0.05) higher in normal cell lines (~50 uM), when compared to that in cancer cell lines (~10 uM). For anti-aging effects, MRC-5 and DSC were exposed to 10 uM EGCG for up to five passages that did not display any growth arrest. Population doubling time and senescence-related ${\beta}-galactosidase$ ($SA-{\beta}-gal$) activity in treated cells were similar to untreated cells. For anti-adipogenic effects, mouse 3T3-L1 pre-adipocytes were induced to adipocytes in an adipogenic differentiation medium containing 10 uM EGCG, but adipogenesis in 3T3-L1 cells was not inhibited by EGCG treatment. For anti-tumor effects, the cancer cell lines were treated with 10 uM EGCG. PDT was significantly (p<0.05) increased in EGCG-treated SNU-601, AGS, MCF-7, and U87-MG cancer cell lines, except in MKN74 and A-549. The level of telomerase activity and cell migration capacity were significantly (p<0.05) reduced, while $SA-{\beta}-gal$ activity was highly up-regulated in EGCG treated-cancer cell lines, when compared to that in untreated cancer cell lines. Our results have demonstrated that EGCG treatment induces anti-tumor effects more efficiently as noted by decreased cell proliferation, cell migration, telomerase activity, and increased $SA-{\beta}-gal$ activity than inducing anti-aging and anti-adipogenesis. Therefore, EGCG at a specific concentration can be considered for a potential anti-tumor drug.

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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Physico-chemical, Nutritional, and Enzymatic Characteristics of Shiitake Spent Mushroom Substrate (SMS) (표고버섯 수확 후 배지의 이화학적, 영양적, 효소적 특성)

  • Sung, Hwa-Jung;Pyo, Su-Jin;Kim, Jong-Sik;Park, Jong-Yi;Sohn, Ho-Yong
    • Journal of Life Science
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    • v.28 no.11
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    • pp.1339-1346
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    • 2018
  • In Korea, edible mushrooms are produced largely on commercial artificial media, so the annual production of spent mushroom substrate (SMS), as a by-product of the mushroom industry, is estimated at over 200 million tons. This SMS is assumed to contain abundant fungal mycelia and pre-fruiting bodies, as well as various nutritive and bioactive compounds that are presently discarded. This study examined the physico-chemical, nutritional, and enzymatic characteristics of uninoculated sterilized medium (USM) and SMS of shiitake mushrooms with the aim of developing a high-value added product from SMS. The contents of crude protein, crude lipid, and ash were higher after the third SMS harvest ($SMS-A-3^{rd}$) than in USM or $SMS-A-1^{st}$. The contents of Ca, Mg, and P in $SMS-A-3^{rd}$ were 2.95, 2.35, and 2.1-fold higher compared than in USM. No As or Cd was detected in USM or SMS. The pH, Brix, and acidity were 4.6, 20.0, and 1.4, respectively in $SMS-A-3^{rd}$, but 5.6, 6.0, and 0.0, respectively, in USM. These results suggest a highly active production of soluble components and organic acids in $SMS-A-3^{rd}$. The distinct color differences noted for USM, $SMS-A-1^{st}$, and $SMS-A-3^{rd}$ could be used as a mycelial growth indicator. Enzyme activity assays using the APIZYM system showed that SMS is a potent source of hydrolysis-related enzymes, especially esterase (C4) and ${\beta}$-glucuronidase. Our results suggested that the SMS of shiitake has a high potential for use in environmental, agricultural, and stock-breeding industries, for example, as active ingredients for sewage treatment, waste-polymer degradation, and feed additives.

Analysis and cloning of cAMP receptor protein(CRp) gene in Serratia marcescens (Serratia marcescens에서 cAMP receptor protein(CRP) 유전자의 클로닝 해석)

  • Yoo, Ju-soon;Kim, Hae-Sun;Moon, Jong-Hwan;Chung, Soo-Yeol;Choi, Yong-Lark
    • Journal of Life Science
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    • v.8 no.3
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    • pp.263-271
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    • 1998
  • One of the better-characterized transcription factor of E. coli is the cAMP receptor protein(CRP) and the CRP binds cAMP and DNA. The cAMP-CRP complex is involved in regulation of many genes at bacteria. The cAMP-CRP regulatory element represents, in some respects, a global regulatory network. The aim of this work was to study the structure and the mechanisms controlling the expression of CRP in Serratia marcescens. We have been get 5 different clones from Serratia which stimulated the cells to use maltose as a sole carbon source in E. coli TP2139. The crp gene clone, pCKB12, was confirmed by Southern hybridization with E. coli crp gene. The location of the crp gene was determined by construction subclones carrying various portions of pCKB12. To investigate the potential role of CRP in E. coli, lacZ fused plasmids were constructed and investigated the ${\beta}$-galactosidase activity of the fused plasmid. The Serratiamarcescens cAMP receptor protein can substitute the E. coli CRP in transcriptional activation at the lacZ gene. These results suggest that Serratia marcescens cAMP receptor protein complex functions to regulate several promoters in E. coli.

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Antimutagenic Mechanism of Water Extract from Rehmannia glutinosa Liboshitz on 4-nitroquinoline 1-oxide Induced Mutagenesis n E. coli B.r (대장균에서 4-nitroquinoline 1-oride의 변이원성에 대한 숙지황 물추출물의 항돌연변이 작용특성)

  • 안병용;한종현;최동성
    • KSBB Journal
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    • v.16 no.5
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    • pp.486-492
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    • 2001
  • The antimutagenic mechanism of the fraction III(RG III)separated from the water extract of Rehmannia glutionosa was investigated by Escherichia. coli GW and B/r strains. RG-III treatment did not affect the ${\beta}$-galactosidase activity E. coli GW-1060, 1106, 1107 and 1105. These results indicated that RG-III did not induce RecA protein amplification and did not also prevent the proteolytic cleavage of LexA. The bio-antimutagenicity and survival effect of RG-III on 4-nitroquinoline 1-oxide(4NQO), N-methyl-N-nitor-N\`-nitrosoguanidine(MNING) were investigate by E. coli B/r strains with have different pathway of DNA repai. RG-III slightly increased the survival of 4NQO-treated WP2, WP2s, WP67, CM561, CM611 cells, but the reactivation of survival cannot ve explained by the repair mode. RG-III caused the decrease of mutagenicity and lethality treated with MNNG in ZA159 despite of the increase in WP2, WP2s, WP67, CW561, CM611. Compared with bio-antimutagenic effects of RG-III on 4NQO, greatly increased antimutagenic effects of RG-III were observed with all the E. coli B/r strains tested, but less active in ZA159. These results suggest that RG-III was identified as a blocking agent for preventing the 4NQO induced mutagenesis, and may act as chl-products.

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Senescence as A Consequence of Ginsenoside Rg1 Response on K562 Human Leukemia Cell Line

  • Liu, Jun;Cai, Shi-Zhong;Zhou, Yue;Zhang, Xian-Ping;Liu, Dian-Feng;Jiang, Rong;Wang, Ya-Ping
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.12
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    • pp.6191-6196
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    • 2012
  • Aims and Background: Traditional chemotherapy strategies for human leukemia commonly use drugs based on cytotoxicity to eradicate cancer cells. One predicament is that substantial damage to normal tissues is likely to occur in the course of standard treatments. Obviously, it is urgent to explore therapies that can effectively eliminate malignant cells without affecting normal cells. Our previous studies indicated that ginsenoside $Rg_1$ ($Rg_1$), a major active pharmacological ingredient of ginseng, could delay normal hematopoietic stem cell senescence. However, whether $Rg_1$ can induce cancer cell senescence is still unclear. Methods: In the current study, human leukemia K562 cells were subjected to $Rg_1$ exposure. The optimal drug concentration and duration with K562 cells was obtained by MTT colorimetric test. Effects of $Rg_1$ on cell cycle were analyzed using flow cytometry and by SA-${\beta}$-Gal staining. Colony-forming ability was measured by colony-assay. Telomere lengths were assessed by Southern blotting and expression of senescence-associated proteins P21, P16 and RB by Western blotting. Ultrastructural morphology changes were observed by transmission electron microscopy. Results: K562 cells demonstrated a maximum proliferation inhibition rate with an $Rg_1$ concentration of $20{\mu}\;mol{\cdot}L^{-1}$ for 48h, the cells exhibiting dramatic morphological alterations including an enlarged and flat cellular morphology, larger mitochondria and increased number of lysosomes. Senescence associated-${\beta}$-galactosidase (SA-${\beta}$-Gal) activity was increased. K562 cells also had decreased ability for colony formation, and shortened telomere length as well as reduction of proliferating potential and arrestin $G_2$/M phase after $Rg_1$ interaction. The senescence associated proteins P21, P16 and RB were significantly up-regulated. Conclusion: Ginsenoside $Rg_1$ can induce a state of senescence in human leukemia K562 cells, which is associated with p21-Rb and p16-Rb pathways.

Functional Characteristics of Kefir as a Fermented Dairy Product: A Review (발효낙농유제품인 Kefir 다양한 기능 및 특성: 총설)

  • Chon, Jung-Whan;Kim, Hyun-Sook;Song, Kwang-Young;Kim, Dong-Hyeon;Kim, Hong-Seok;Yim, Jin-Hyeok;Choi, Dasom;Hwang, Dae-Geun;Kim, Young-Ji;Lee, Soo-Kyung;Seo, Kun-Ho
    • Journal of Dairy Science and Biotechnology
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    • v.31 no.2
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    • pp.99-108
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    • 2013
  • Kefir is a unique fermented dairy product resulting from combined lactic acid and alcoholic fermentation of lactose in milk. Kefir is produced by the microbial activity of kefir grains. Kefir has numerous health benefits such as the reduction of cholesterol levels, stimulation of the immune system, antimutagenic and anticarcinogenic properties, and improvements of lactose intolerance. Furthermore, kefir is excellent as both a dietetic beverage and for protection against various diseases in small babies. Therefore, kefir has recently been regarded as an important functional dairy food. To date, the majority of research on kefir has focused on the applications of functional kefir using advanced biotechnology methods. The purpose of this review article is to facilitate the recognition of kefir as a novel functional food.

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