• Title/Summary/Keyword: Extracellular

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Evaluation of Achyranthes japonica Ethanol Extraction on the Inhibition Effect of Hyluronidase and Lipoxygenase (쇠무릎 에탄올 추출물의 DPPH, 히알루로니다아제 및 리폭시게나아제 저해 효과)

  • Cho, Kyung-Soon
    • Journal of Life Science
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    • v.25 no.12
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    • pp.1370-1376
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    • 2015
  • The 1, 1- diphenyl 2-picrylhyorazyl (DPPH) is a well-known radical and a trap (scavenger) for other radicals. Hyaluronidase (HAase) is an enzyme that depolymerizes the polysaccharide hyaluronic acid (HA) in the extracellular matrix of connective tissue. Lipoxygenase (LOX) enzyme was reported to convert the arachidonic, linoleic and other polyunsaturated fatty acid into biologically active metabolites involved in the inflammatory and immune responses. The purpose of the present study is to evaluate plant extracts as sources of natural antioxidants and to examine whether Achyranthes japonica having significant DPPH, HAase and LOX inhibitory activity. The inhibitory effect of HAase by A. japonica was assayed using a Morgan microplate assay. The antioxidant activity of the A. japonica extracts was measured on the basis of the scavenging activity of the stable 1, 1- diphenyl 2-picrylhyorazyl (DPPH) free radical. DPPH scavenging activity of matured roots of A. japonica was evaluated at 4.0 mg/ml was 87.8% and that of young roots was 86.2% at same concentration. The roots of A. japonica showed maximum inhibition of HAase activity (IC50 = 27.7 μg/ml). The highest LOX inhibition was recorded in the root extract among three vegetative parts. Inhibition of HAase activity of roots may contribute towards the development of herbal medicines. Although percent inhibition of lipoxygenase by Achyranthes japonica for all young and matured groups for leaves, stems, and roots at different concentrations, there were not show a statistically significant difference (p<0.05).

Optimal Culture Conditions on the Keratinase Production by Bacillus sp. SH-517. (Bacillus sp. SH-517에 의한 keratinase의 생성 최적 배양 조건)

  • Bang, Byung-Ho;Rhee, Moon-Soo;Lim, Ki-Hwan;Yi, Dong-Heui
    • Journal of Life Science
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    • v.18 no.6
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    • pp.839-844
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    • 2008
  • A strain SH-517 which produce extracellular keratinase, was isolated from the soil of a poultry waste and a poultry factory. An isolate SH-517 was identified as Bacillus sp. based on its morphological and biochemical characteristics. The optimal culture conditions for the production of keratinase by Bacillus sp. SH-517 were investigated. The optimal medium composition for keratinase production was determined to be 2.0% chicken feather as carbon source, 0.5% beef extract as organic nitrogen source, 0.5% $KNO_3$ as inorganic nitrogen source and 0.06% KCl, 0.05% NaCl, 0.04% $KH_2PO_4$, 0.03% $K_2HPO_4$ as mineral source and 0.01% yeast extract as growth factor. The optimal temperature and pH of medium were shown $40^{\circ}C$ and 8.5 with shaking culture (180 rpm/min), respectively. The maximum keratinase production reached maximum of 125 units/ml/min after 42 hr of cultivation under the optimal culturing conditions.

Bacterial Community of Natural Dye Wastewater Treatment Facility (천연염색 폐수처리시설의 세균 군집)

  • Hwang, Yeoung Min;Kim, Dae Kuk;Lee, Ji Hee;Baik, Keun Sik;Park, Chul;Seong, Chi Nam
    • Journal of Life Science
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    • v.24 no.4
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    • pp.393-402
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    • 2014
  • Culture-dependent and culture-independent denaturing gradient gel electrophoresis (DGGE) analyses were employed to investigate the bacterial community associated with a natural dye wastewater treatment facility. A total of 104 (influent water, 48 strains; aeration tank, 25; settling tank, 31) bacterial strains were isolated. Based on the 16S rRNA gene sequences comparison analysis, the isolates belonged to four phyla: Proteobacteria, Actinobacteria, Firmicutes, and Bacteriodetes. Seventeen DGGE bands representing dominant taxa in each sample were cloned and partially sequenced. The same four phyla were detected by DGGE fingerprinting. The most dominant taxon retrieved by both methods was the member of the phylum Proteobacteria with Alphaproteobacteria as the predominant class. The bacterial community associated with the natural dye wastewater treatment facility is composed of parasites of animals and plants, decomposers of polysaccharides and dyes, and producers of extracellular polysaccharides.

The Optimal Condition and Enzyme Activity of Entomopathogenic Fungus Beauveria bassiana Using Extracted Rice Bran (미강추출물을 이용한 곤충병원성 곰팡이 Beauveria bassiana의 최적 배양조건 및 효소활성)

  • Kim, Chang-Su;Lee, Jung-Bok;Kim, Beam-Soo;Lee, Min-Hye;Kang, Kyeong-Muk;Joo, Woo-Hong;Kim, Jin-Won;Im, Dae-Joon;Kwon, Gi-Seok
    • Journal of Life Science
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    • v.23 no.8
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    • pp.1010-1018
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    • 2013
  • The greenhouse whitefly, Bemisia tabaci, is considered one of the most destructive pests of crops. In this study, we aimed to determine the optimal liquid culture conditions in shake flasks for maximal sporulation of Beauveria bassiana M130 using rice bran. The optimal initial pH for the spore production of B. bassiana using extracted rice bran medium was 5.2 and $28^{\circ}C$. The screening in shake flasks of carbon and nitrogen sources resulted in the identification of an optimal medium based on 0.5% $(NH_4)_2SO_4$, with extracted rice bran 8:1. Using this medium, a production level of $2.15{\times}10^9$ spores per ml was obtained after six days from culture inoculation at $28^{\circ}C$ in a rotary shaking incubator at 130 rpm. In addition, the specific activities of extracellular enzymes of chitinase and protease were $4,296{\mu}mol$ and $375{\mu}mol$, respectively. These results suggest that Beauveria bassiana M130 could be a bio-controller for the greenhouse whitefly.

Effects of Transient Treatment with Rotenone, a Mitochondrial Inhibitor, on Mouse Subventricular Zone Neural Stem Cells (미토콘드리아 저해제인 rotenone의 일시적 처리가 쥐의 뇌실 하 영역 신경 줄기 세포에 미치는 영향)

  • Park, Ki-Youb;Kim, Man Su
    • Journal of Life Science
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    • v.29 no.12
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    • pp.1329-1336
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    • 2019
  • Subventricular zone (SVZ) in the brain contains neural stem cells (NSCs) which self-renew and differentiate to neurons and glial cells during postnatal period and throughout adulthood. Since fate decision to either proliferation or differentiation has to respond to intracellular and extracellular conditions, many intrinsic and extrinsic factors are involved. Among them, mitochondria have been reported to participate in fate decision of NSCs. In our previous report, we showed that long-term treatment of a mitochondrial inhibitor rotenone greatly inhibited neurogenesis. In this study, we examined the effects of short-term treatment of rotenone on SVZ NSCs. We found that (1) even one-day treatment of rotenone significantly reduced neurogenesis and earlier time points seemed to be more sensitive to rotenone, (2) a number of Mash1+ transit amplifying cells was decreased by one-day treatment of rotenone, (3) short-term treatment of rotenone eliminated most of the differentiated Tuj1+ neurons and Olig2+ oligodendrocytes, while glial fibrillary acidic protein (GFAP)+ astrocytes were not affected, and (4) sulfiredoxin 1 (Srxn1) gene expression was increased after one-day treatment of rotenone, indicating activation of nuclear factor (erythroid-derived 2)-like 2 (Nrf2) pathway. All these results confirm that functional mitochondria are necessary during differentiation to neurons or oligodendrocytes as well as maintenance of neurons after differentiation. Also, these data suggest that temporary exposure to mitochondrial inhibitor such as rotenone might have long-term effects on neurogenic potential of NSCs.

MiR-29a and MiR-140 Protect Chondrocytes against the Anti-Proliferation and Cell Matrix Signaling Changes by IL-1β

  • Li, Xianghui;Zhen, Zhilei;Tang, Guodong;Zheng, Chong;Yang, Guofu
    • Molecules and Cells
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    • v.39 no.2
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    • pp.103-110
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    • 2016
  • As a degenerative joint disease, osteoarthritis (OA) constitutes a major cause of disability that seriously affects the quality of life of a large population of people worldwide. However, effective treatment that can successfully reverse OA progression is lacking until now. The present study aimed to determine whether two small non-coding RNAs miR-29a and miR-140, which are significantly down-regulated in OA, can be applied together as potential therapeutic targets for OA treatment. MiRNA synergy score was used to screen the miRNA pairs that potentially synergistically regulate OA. An in vitro model of OA was established by treating murine chondrocytes with IL-$1{\beta}$. Transfection of miR-29a and miR-140 via plasmids was investigated on chondrocyte proliferation and expression of nine genes such as ADAMTS4, ADAMTS5, ACAN, COL2A1, COL10A1, MMP1, MMP3, MMP13 and TIMP metallopeptidase inhibitor 1 (TIMP1). Western blotting was used to determine the protein expression level of MMP13 and TIMP1, and ELISA was used to detect the content of type II collagen. Combined use of miR-29a and miR-140 successfully reversed the destructive effect of IL-$1{\beta}$ on chondrocyte proliferation, and notably affected the MMP13 and TIMP1 gene expression that regulates extracellular matrix. Although co-transfection of miR-29a and miR-140 did not show a synergistic effect on MMP13 protein expression and type II collagen release, but both of them can significantly suppress the protein abundance of MMP13 and restore the type II collagen release in IL-$1{\beta}$ treated chondrocytes. Compared with single miRNA transfection, cotransfection of both miRNAs exceedingly abrogated the suppressed the protein production of TIMP1 caused by IL-$1{\beta}$, thereby suggesting potent synergistic action. These results provided1novel insights into the important function of miRNAs' collaboration in OA pathological development. The reduced MMP13, and enhanced TIMP1 protein production and type II collagen release also implies that miR-29a and miR-140 combination treatment may be a possible treatment for OA.

Histopathological studies on the macrophage behavior in lymphomyeloid tissues of tilapia, Oreochromis niloticus (나일틸라피아의 임파조혈조직내 대식세포(大食細胞) 분포변화(分布變化)에 관한 병리조직학적(病理組織學的) 연구(硏究))

  • Lee, Nam-Sil;Kim, Jee-Young;Jeong, Hyun-Do;Huh, Min-Do
    • Journal of fish pathology
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    • v.8 no.2
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    • pp.135-148
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    • 1995
  • To elucidate the distributional pattern of macrophages within lymphomyeloid tissues according to the disease process, tilapias, teleostean fish, were intraperitoneally injected with live Edwardsiellatarda and its extracellular product(ECP) respectively. And then histopathological examination for the spleen and gead kidney were carried out for the individuals which had not any clinical signs. In the group injected with live E. tarda, macrophages were densely organized into MMC-like structures with showing some degree of recovery in histological arrangement. At the 2nd week, overall structures of the lymphomyeloid tissues became normal, accompanying the disappearance of most of macrophage groups. Also in case of ECP injection, quite similar findings were observed. Moreover, macrophage collections and hypertrophied ellipsoids were recognized at 1hour after the injection of ECP in head kidney and spleen, respectively. These results suggested that characteristic behaviors of macrophages in lymphomyeloid tissues would be used as important morphological criteria for early diagnosis of edwarsiellosis or possibly of other infectious diseases.

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KCl Mediates $K^+$ Channel-Activated Mitogen-Activated Protein Kinases Signaling in Wound Healing

  • Shim, Jung Hee;Lim, Jong Woo;Kim, Byeong Kyu;Park, Soo Jin;Kim, Suk Wha;Choi, Tae Hyun
    • Archives of Plastic Surgery
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    • v.42 no.1
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    • pp.11-19
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    • 2015
  • Background Wound healing is an interaction of a complex signaling cascade of cellular events, including inflammation, proliferation, and maturation. $K^+$ channels modulate the mitogen-activated protein kinase (MAPK) signaling pathway. Here, we investigated whether $K^+$ channel-activated MAPK signaling directs collagen synthesis and angiogenesis in wound healing. Methods The human skin fibroblast HS27 cell line was used to examine cell viability and collagen synthesis after potassium chloride (KCl) treatment by Cell Counting Kit-8 (CCK-8) and western blotting. To investigate whether $K^+$ ion channels function upstream of MAPK signaling, thus affecting collagen synthesis and angiogenesis, we examined alteration of MAPK expression after treatment with KCl (channel inhibitor), NS1619 (channel activator), or kinase inhibitors. To research the effect of KCl on angiogenesis, angiogenesis-related proteins such as thrombospondin 1 (TSP1), anti-angiogenic factor, basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF), pro-angiogenic factor were assayed by western blot. Results The viability of HS27 cells was not affected by 25 mM KCl. Collagen synthesis increased dependent on time and concentration of KCl exposure. The phosphorylations of MAPK proteins such as extracellular-signal-regulated kinase (ERK) and p38 increased about 2.5-3 fold in the KCl treatment cells and were inhibited by treatment of NS1619. TSP1 expression increased by 100%, bFGF expression decreased by 40%, and there is no significant differences in the VEGF level by KCl treatment, TSP1 was inhibited by NS1619 or kinase inhibitors. Conclusions Our results suggest that KCl may function as a therapeutic agent for wound healing in the skin through MAPK signaling mediated by the $K^+$ ion channel.

Usefulness of Atelo-collgen sponge (Teruplug$^{(R)}$) for Treatment of Mandibular Angle Fractures with Third Molar Extraction (하악각 골절에서 제 3 대구치 발치 후 아테로-콜라겐 스펀지의 유용성)

  • Oh, Hwa-Young;Choi, Hwan-Jun;Kwon, Jun-Seong;Lee, Hyung-Gyo;Kim, Yong-Bae
    • Archives of Plastic Surgery
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    • v.38 no.2
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    • pp.155-160
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    • 2011
  • Purpose: Fibrillar collagens like type I collagen, are the major constituent of the extracellular matrix and structural protein of bone. Also, it can be a scaffold for osteoblast migration. The purpose of this study is to estimate the effects of absorbable atelo-collagen sponge (Teruplug$^{(R)}$, Terumo biomaterials Co., Tokyo, Japan) insertion in tooth extraction sites on periodontal healing of the second molar, healing of the fractured mandibular bone and new bone formation of third molar socket after the extraction of the impacted third molar with mandibular angle fracture. Methods: In our study of six cases of mandibular angle fractures, all of them underwent the extraction of the third molar tooth & absorbable atelo-collagen sponge insertion in tooth extraction site. Three of them had a intraoral infection & oral opening to fracture site, two of the six had dental caries, and only one had reduction problem due to third molar position. Six consecutive patients with noncomminuted fractures of the mandibular angle were treated by open reduction and internal fixation using one noncompression miniplates and screws placed through a transoral incision. Results: All of the patients have showed good postoperative functions and have not experienced complications requiring second surgical intervention. There was well healing of the mandibular bone and the most new bone formation of third molar socket after the extraction of the impacted third molar with mandibular angle fracture. Conclusion: The results of this study suggest that absorbable atelo-collagen sponge is relatively favorable bone void filler with prevention of tissue collapse, food packing, and enhance periodontal healing. Thus, the use of atelo-collagen sponge and one noncompression miniplate seems to be relatively easy, safe, and effective for the treatment of fractures of the mandibular angle and third molar extraction.

The Activation of Stress-induced Heat Shock Protein 27 and the Relationship of Physical Therapy (스트레스-유도 열충격단백질 27(Heat Shock Protein 27)의 활성과 물리치료의 상관성)

  • Kim, Mi-Sun;Lee, Sung-Ho;Kim, Il-Hyun;Hwang, Byong-Yong;Kim, Jung-Hwan
    • The Journal of Korean Physical Therapy
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    • v.20 no.1
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    • pp.57-65
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    • 2008
  • Purpose: Heat shock proteins (HSPs) are a group of proteins that are activated when cells are exposed to a variety of environmental stresses, such as infection, inflammation, exposure to toxins, starvation, hypoxia, brain injury, or water deprivation. The activation of HSPs by environmental stress plays a key role in signal transduction, including cytoprotection, molecular chaperone, anti-apoptotic effect, and anti-aging effects. However, the precise mechanism for the action of small HSPs, such as HSP27 and mitogen-activated protein kinases (MAPKs: extracellular-regulated protein kinase 1/2 (ERK1/2), p38MAPK, stress-activated protein kinase/c-Jun N-terminal kinase (SAPK/JNK), is not completely understood, particularly in application of cell stimulators including platelet-derived growth factor (PDGF), angiotensin II (AngII), tumor necrosis factor $\alpha$ (TNF$\alpha$), and $H_2O_2$. This study examined the relationship between stimulators-induced enzymatic activity of HSP27 and MAPKs from rat smooth and skeletal muscles. Methods: 2-dimensional electrophoresis (2DE) and matrix assisted laser desorption ionizationtime-of-flight/time-of-flight (MALDI-TOF/TOF) analysis were used to identify HSP27 from the intact vascular smooth and skeletal muscles. Three isoforms of HSP27 were detected on silver-stained gels of the whole protein extracts from the rat aortic smooth and skeletal muscle strips. Results: The expression of PDGF, AngII, TNF$\alpha$, and $H_2O_2$-induced activation of HSP27, p38MAPK, ERK1/2, and SAPK/JNK was higher in the smooth muscle cells than the control. SB203580 (30${\mu}$M), a p38MAPK inhibitor, increased the level of HSP27 phosphorylation induced by stimulators in smooth muscle cells. Furthermore, the age-related and starvation-induced activation of HSP27 was higher in skeletal muscle cells (L6 myoblast cell lines) and muscle strips than the control. Conclusion: These results suggest, in part, that the activity of HSP27 and MAPKs affect stressors, such as PDGF, AngII, TNF$\alpha$, $H_2O_2$, and starvation in rat smooth and skeletal muscles. However, more systemic research will be needed into physical therapy, including thermotherapy, electrotherapy, radiotherapy and others.

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