• Title/Summary/Keyword: C3H/10T1/2 cells

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A Mophological Study on the Cytotoxic Effect of Nicotine in Mice Testis (Nicotine이 생쥐 고환에 미치는 세포독성효과)

  • Kim, C.-H.;Koh, P.-O.;Won, C.-K.;Kim, J.-S.;Kang, C.-B.;Kang, My.-G.;Kim, T.-S.;Gal, G.-Y.;Chung, J.-Y.;Park, H.-S.
    • Journal of Embryo Transfer
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    • v.19 no.2
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    • pp.75-80
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    • 2004
  • This study was performed to examine the cytotoxic effects of nicotine on the seminiferous tubules and Leydig cells in mice testis. A different amount of nicotine (2~15 mg/kg, for two weeks, one dose of 100 mg/kg) were administered to four-month male mice, and then the optical microscopic findings of its effect on testis of the mice are as follows: 1. The group that 2 mg/kg of nicotine was administered showed normal findings that nucleus and cytoplasm of Leydig cells are distinct, while the other group that 5 mg/kg of nicotine was given to showed nucleus and cytoplasm are swollen and thickened a little, and slightly dyed. 2. The group that 10 mg/kg of nicotine was given had irregular arrangement of spermatogenesis inside seminiferous tubules so it was impossible to distinguish phages of seminiferous tubules. It was also impossible to observe cells due to fusion of their nucleuses, and distinct cytoplasm. 3. The group that 15 mg/kg of nicotine was administered showed destruction of nucleuses and cytoplasm of spermatocytes and sperms and a fill of fibered connective tissues so that it is impossible to observe rumens of seminiferous tubules.

A Novel Metalloprotease from the Wild Basidiomycete Mushroom Lepista nuda

  • Wu, Y.Y.;Wang, H.X.;Ng, T.B.
    • Journal of Microbiology and Biotechnology
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    • v.21 no.3
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    • pp.256-262
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    • 2011
  • A 20.9-kDa metalloprotease was isolated from dried fruiting bodies of the wild basidiomycete mushroom Lepista nuda. The N-terminal amino acid sequence of the protease was seen to be ATFVLTAATNTLFTA, thus displaying no similarity with the sequences of previously reported metalloproteases. The protease was purified using a procedure that entailed ion-exchange chromatography on CM-Cellulose, Q-Sepharose, and Mono S, and FPLC-gel filtration on Superdex 75. The protease functioned at an optimum pH of 7.0 and an optimum temperature of $50^{\circ}C$. It was also noted that the protease demonstrated a proteolytic activity of 1,756 U/mg toward casein. The $K_m$ of the purified protease toward casein was 6.36 mg/ml at a pH of 7.0 and with a temperature of $37^{\circ}C$, whereas the $V_{max}$ was 9.11 ${\mu}g\;ml^{-1}\;min^{-1}$. The activity of the protease was adversely affected by EDTA-2Na, suggesting that it is a metalloprotease. PMSF, EGTA, aprotinin, and leupeptin exerted no striking inhibitory effect. The activity of the protease was enhanced by $Fe^{2+}$, but was curtailed by $Cd^{2+}$, $Cu^{2+}$, $Hg^{2+}$, $Pb^{2+}$, $Zn^{2+}$, and $Fe^{2+}$ ions. The protease also exhibited inhibitory activity against HIV-1 reverse transcriptase with an $IC_{50}$ value of 4.00 ${\mu}M$. The $IC_{50}$ values toward hepatoma Hep G2 and leukemia L1210 cells in vitro were 4.99 ${\mu}M$ and 3.67 ${\mu}M$, respectively.

Effects of Danggwisayeokgaohsuyusaenggang-tang(當歸四逆加吳茱萸生薑湯) on Collagen-induced Arthritis in Mice (Collagen으로 유발된 생쥐의 관절염에 대한 당귀사역가오수유생강탕(當歸四逆加吳茱萸生薑湯)의 효과)

  • Kim, Min-Kyun;Oh, Min-Seok
    • Journal of Korean Medicine Rehabilitation
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    • v.21 no.2
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    • pp.63-85
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    • 2011
  • Objectives : This study was carried out to know the effects of Danggwisayeokgaohsuyusaenggang-tang(hereinafter referred to DST) on arthritis induced by collagen on DBA/1 OlaHsd mice. Methods : For this purpose, DST was orally administered to mouse with arthritis induced by collagen II. Cytotoxicity, high performance liquid chromatograph(HPLC) analysis, arthritis index, value of immunocyte in draining lymph node and paw joint, cytokine were measured in vivo. Results : 1. The cytotoxicity against human fibroblast cells(hFCs) was not measured in any concentration. 2. In HPLC analysis, There are high peak patterns at 8 minute(min), 12 min, 35 min, 45 min. 3. The arthritis index was decreased significantly. 4. The degree of arthritis induced damage of joint of DST group is slight compared with control group in histopathologic observation(Hematoxylin and eosin stain(H&E), Masson's trichrome(M-T) staining). 5. In total cell counts of draining lymph node(DLN) and paw joint, the cells in DLN decreased significantly on DST 200 mg/kg and the cells in paw joint decreased significantly on 200 mg/kg and 50 mg/kg. 6. In DLN, $CD4^+/CD25^+$, $CD3^+/CD69^+$, major histocompatibility complex(MHC), class-II/$CD11c^+$ cells decreased significantly on DST 200 mg/kg and 50 mg/kg $CD3^+/CD8^+$ cells decreased significantly on DST 200 200 mg/kg, $CD4^+$, $CD3^+/CD44^+$ cells decreased. 7. In paw joints, $CD4^+$, $CD11b^+/Gr-1^+$ cells decreased significantly on DST 200 mg/kg and 50 mg/kg. 8. In joints, levels of $IL-1{\beta}$, IL-6, $TNF-{\alpha}$, cyclo-oxygenase-2(COX-2), NOS-II were decreased on DST 200 mg/kg and DST 50 mg/kg. 9. In analysing of cytokine in CD3/CD28 activated spleen, IL-17 was decreased significantly, IL-4 was increased significantly $INF-{\gamma}$ was decreased on DST 200 mg/kg. 10. In analysing of cytokine in collagen activated spleen, IL-17 were decreased significantly, IL-4 was increased significantly. Conclusions : This results demonstrated that DST suppressed the inflammatory progression of collagen-induced arthritis(CIA) mice and supported further studies are required to survey continuously in looking for the effective substance and mechanism in the future.

Tunicamycin negatively regulates BMP2-induced osteoblast differentiation through CREBH expression in MC3T3E1 cells

  • Jang, Won-Gu;Kim, Eun-Jung;Koh, Jeong-Tae
    • BMB Reports
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    • v.44 no.11
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    • pp.735-740
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    • 2011
  • Tunicamycin, an endoplasmic reticulum (ER) stress inducer, specifically inhibits N-glycosylation. The cyclic AMP (cAMP) response element-binding protein H (CREBH) was previously shown to be regulated by UPR-dependent proteolytic cleavage in the liver. On the other hand, the role of CREBH in other tissues is unknown. In the present study, tunicamycin increased the level of CREBH activation (cleavage) as well as mRNA expression in osteoblast cells. Adenoviral (Ad) overexpression of CREBH suppressed BMP2-induced expression of alkaline phosphatase (ALP) and osteocalcin (OC). Interestingly, the BMP2-induced OASIS (structurally similar to CREBH, a positive regulator of osteoblast differentiation) expression was also inhibited by CREBH overexpression. In addition, inhibition of CREBH expression using siRNA reversed the tunicamycin-suppressed ALP and OC expression. These results suggest that CREBH inhibited osteoblast differentiation via suppressing BMP2-induced ALP, OC and OASIS expression in mouse calvarial derived osteoblasts.

Corrosion Characteristics of Cell-Covered Ternary Ti-Nb-Ta Alloy for Biomaterials

  • Kim, W.G.;Yu, J.W.;Choe, H.C.;Ko, Y.M.;Park, G.H.
    • Corrosion Science and Technology
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    • v.8 no.2
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    • pp.62-67
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    • 2009
  • Ti and Ti-alloys have good biocompatibility, appropriate mechanical properties and excellent corrosion resistance. However, the widely used Ti-6Al-4V is found to release toxic ions (Al and V) into the body, leading to undesirable long-term effects. Ti-6Al-4V has much higher elastic modulus (100 GPa) than cortical bone (20 GPa). Therefore, titanium alloys with low elastic modulus have been developed as biomaterials to minimize stress shielding. The electrochemical behavior of surface-modified and MC3T3-E1 cell-cultured Ti-30(Nb,Ta) alloys with low elastic modulus have been investigated using various electrochemical methods. Surfaces of test samples were treated as follows: $0.3{\mu}m$ polished; $25{\mu}m$, $50{\mu}m$ and $125{\mu}m$ sandblasted. Specimen surfaces were cultured with MC3T3-E1 cells for 2 days. Average surface roughness ($R_a$) and morphology of specimens were determined using a surface profilometer, OM, and FE-SEM. Corrosion behavior was investigated using a potentiostat(EG&G PARSTAT 2273), and electrochemical impedance spectroscopy was performed (10 mHz to 100 kHz) in 0.9% NaCl solution at $36.5{\pm}1^{\circ}C$. The microstructures of the Ti-30(Ta,Nb) alloys had a needle-like appearance. The $R_a$ of polished Ti-30Ta and Ti-30Nb alloys was lower than that of the sandblasted Ti alloy. Cultured cells displayed round shapes. For polished alloy samples, cells were well-cultured on all surfaces compared to sandblasted alloy samples. In sandblasted and cell-cultured Ti-30(Nb,Ta) alloy, the pitting potential decreased and passive current density increased as $R_a$ increased. Anodic polarization curves of cell-cultured Ti alloys showed unstable behavior in the passive region compared to non-cell-cultured alloys. From impedance tests of sandblasted and cell-cultured alloys, the polarization resistance decreased as $R_a$ increased, whereas, $R_a$ for cell-cultured Ti alloys increased compared to non-cell-cultured Ti alloys.

Acinetobacter pullorum sp. nov., Isolated from Chicken Meat

  • Elnar, Arxel G.;Kim, Min-Gon;Lee, Ju-Eun;Han, Rae-Hee;Yoon, Sung-Hee;Lee, Gi-Yong;Yang, Soo-Jin;Kim, Geun-Bae
    • Journal of Microbiology and Biotechnology
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    • v.30 no.4
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    • pp.526-532
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    • 2020
  • A bacterial strain, designated B301T and isolated from raw chicken meat obtained from a local market in Korea, was characterized and identified using a polyphasic taxonomic approach. Cells were gram-negative, non-motile, obligate-aerobic coccobacilli that were catalase-positive and oxidase-negative. The optimum growth conditions were 30℃, pH 7.0, and 0% NaCl in tryptic soy broth. Colonies were round, convex, smooth, and cream-colored on tryptic soy agar. Strain B301T has a genome size of 3,102,684 bp, with 2,840 protein-coding genes and 102 RNA genes. The 16S rRNA gene analysis revealed that strain B301T belongs to the genus Acinetobacter and shares highest sequence similarity (97.12%) with A. celticus ANC 4603T and A. sichuanensis WCHAc060041T. The average nucleotide identity and digital DNA-DNA hybridization values for closely related species were below the cutoff values for species delineation (95-96% and 70%, respectively). The DNA G+C content of strain B301T was 37.0%. The major respiratory quinone was Q-9, and the cellular fatty acids were primarily summed feature 3 (C16:1 ω6c/C16:1 ω7c), C16:0, and C18:1 ω9c. The major polar lipids were phosphatidylethanolamine, diphosphatidyl-glycerol, phosphatidylglycerol, and phosphatidyl-serine. The antimicrobial resistance profile of strain B301T revealed the absence of antibiotic-resistance genes. Susceptibility to a wide range of antimicrobials, including imipenem, minocycline, ampicillin, and tetracycline, was also observed. The results of the phenotypic, chemotaxonomic, and phylogenetic analyses indicate that strain B301T represents a novel species of the genus Acinetobacter, for which the name Acinetobacter pullorum sp. nov. is proposed. The type strain is B301T (=KACC 21653T = JCM 33942T).

Chemical Composition and Antitumor Apoptogenic Activity of Methylene Chloride Extracts from the Leaves of Zanthoxylum schinifolium (Zanthoxylum schinifolium잎의 methylene chloride 추출물의 화학적 조성 및 암세포에 대한 세포자살 유도활성과 그 작용기전)

  • Kim Jun-Seok;Jun Do-Youn;Woo Mi-Hee;Rhee In-Koo;Kim Young-Ho
    • Journal of Life Science
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    • v.16 no.3 s.76
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    • pp.546-554
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    • 2006
  • To understand antitumor activity of Zanthoxylum schinfolium, which has been used as an aromatic and medicinal plant in Korea, the cytotoxic effect of various organic solvent extracts of its leaves on human tumor cells were investigated. Among these extracts such as methanol extract (SL-13), methylene chloride extract (SL-14), ethyl acetate extract (SL-15), n-butanol extract (SL-16), and residual fraction (SL-17), SL-14 appeared to contain the most cytotoxic activity against leukemia and breast cancer cells tested. The methylene chloride extra.1 (SL-14) possessed an apoptogenic activity causing apoptotic DNA fragmentation of human acute leukemia Jurkat T cells via mitochondrial cytochrome c release into cytoplasm, subsequent activation of caspase-9 and caspase-3, and cleavage of PARP, which could be negatively regulated by antiapoptotic protein Bcl-xL. The GC-MS analysis of SL-14 revealed that the twenty-two ingredients of SL-14 were 9,19-cyclolanost-24-en-3-ol (15.1%), 2-a-methyl-17, b-hop-21-ene (15.1%), 15-methyl-2,3-dihydro-1H benzazepin (11.95%), phytol (10.38%), lupeol (9.92%), 12-methylbenzofuran (8.23%), hexadecanoic acid (5.96%), cis,cis,cis-9,12,15-octadecatrienoic acid-methyl-ester (5.49%), 9,12,15-octadecatrienoic acid-methylester (3.59%), 15-methyl-4-(1-methylethylidene)-2-(4-nitrophenyl) (3.36%), hexadecanoic acid methyl ester (1.93%), vitamine E (1.88%), beta-amyrin (0.96%), and auraptene (0.89%). These results demonstrate that the cytotoxicity of the methylene chloride extract of the leaves of Z. schinifolium toward Jurkat T cells is mainly attributable to apoptosis mediated by mitochondria-dependent caspase cascade regulated by Bcl-xL, and provide an insight into the mechanism underlying antitumor activity of the edible plant Z. schinifolium.

Fabrication and Characteristics of PIN Type Amorphous Silicon Solar Cell (PIN形 非晶質 硅素 太陽電池의 製作 및 特性)

  • Park, Chang-Bae;Oh, Sang-Kwang;Ma, Dae-Yeong;Kim, Ki-Wan
    • Journal of the Korean Institute of Telematics and Electronics
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    • v.26 no.6
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    • pp.30-37
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    • 1989
  • The PIN type a-SiC:H/a-Si:H heterojunction solar cells were fabricated by using the rf glow discharge decomposition of $SiH_4$ mixed with $CH_4,B_2,H_6\;and\;PH_3.$ The efficiency of the solar cell of the $SnO_2/ITO$ was higher than that of ITO transparent oxide layer by 1.5%. The P layer was prepared with the thickness of $100{\AA}$ and $CH_4/SiH_4$ ration of 5. The I layer has been deposited on the P layer and it is not pure intrinsic but near N type. So $SiH_4$ mixed with $B_2H_6$ of 0.3ppm was used to change this N type nature to intrinsic having the thickness of 5000${\AA}$. And consecutively, the N layer was deposited with t ethickness of $400{\AA}$ using $SiH_4/PH_3$ mixtures. The solar cell demonstrated 0.94V of $V_{oc'}$ 14.6mA/cm of $J_{sc}$ and 58.2% of FF, resulting the efficiency of 8.0%. To minimize loss by the reflection of light, $MgF_2$ layer was coated on the lgass and the efficiency was improved by 0.5%. Therefore, the solar cell indicated overall efficiency of 8.5%.

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Anticancer Activity of Extremely Effective Recombinant L-Asparaginase from Burkholderia pseudomallei

  • Darwesh, Doaa B.;Al-Awthan, Yahya S.;Elfaki, Imadeldin;Habib, Salem A.;Alnour, Tarig M.;Darwish, Ahmed B.;Youssef, Magdy M.
    • Journal of Microbiology and Biotechnology
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    • v.32 no.5
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    • pp.551-563
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    • 2022
  • L-asparaginase (E.C. 3.5.1.1) purified from bacterial cells is widely used in the food industry, as well as in the treatment of childhood acute lymphoblastic leukemia. In the present study, the Burkholderia pseudomallei L-asparaginase gene was cloned into the pGEX-2T DNA plasmid, expressed in E. coli BL21 (DE3) pLysS, and purified to homogeneity using Glutathione Sepharose chromatography with 7.26 purification fold and 16.01% recovery. The purified enzyme exhibited a molecular weight of ~33.6 kDa with SDS-PAGE and showed maximal activity at 50℃ and pH 8.0. It retained 95.1, 89.6%, and 70.2% initial activity after 60 min at 30℃, 40℃, and 50℃, respectively. The enzyme reserved its activity at 30℃ and 37℃ up to 24 h. The enzyme had optimum pH of 8 and reserved 50% activity up to 24 h. The recombinant enzyme showed the highest substrate specificity towards L-asparaginase substrate, while no detectable specificity was observed for L-glutamine, urea, and acrylamide at 10 mM concentration. THP-1, a human leukemia cell line, displayed significant morphological alterations after being treated with recombinant L-asparaginase and the IC50 of the purified enzyme was recorded as 0.8 IU. Furthermore, the purified recombinant Lasparaginase improved cytotoxicity in liver cancer HepG2 and breast cancer MCF-7 cell lines, with IC50 values of 1.53 and 18 IU, respectively.

Invitro and Virtual Screening of Bioactive Molecule from Mycelium of Trichoderma atroviride Inhibit the UDP-3-O-(R-3-hydroxymyristoyl)-N-acetylglucosamine Deacetylases (LpxC) for Treatment of Bacterial Infection

  • Saravanakumar, Kandasamy;Park, Cheol-Ho;Wang, Myeong-Hyeon
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2018.04a
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    • pp.67-67
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    • 2018
  • Trichoderma species are a rich source of metabolites, but less known for biomedical potential. This work deals with antibacterial and antioxidant potentials of intracellular non-cytotoxic metabolites, extracted from Trichoderma atroviride (KNUP001). A total of 53 fractions was collected by column chromatography and tested for cytotoxicity by MTT assay. Only one fraction (F41) was found to be non-toxic to Vero cells with $95.4{\pm}0.61%$ of survival. The F41 was then subjected to chemical analysis, antibacterial and antioxidant assays. The F41 at $500{\mu}g.ml^{-1}$ showed the total antioxidant of $48.70{\pm}2.90%$, DPPH radical scavenging activity of $37.25{\pm}2.25$, nitric oxide (NO) radical scavenging activity of $54.55{\pm}1.95$ and $H_2O_2$ radical scavenging activity of $43.75{\pm}3.21$. The F41 at $25{\mu}g.ml^{-1}$ displayed antibacterial activity against E. coli ($14.25{\pm}0.2mm$), P. mirabilis ($10.4{\pm}0.6mm$), S. dysenteriae ($18.6{\pm}03mm$), S. paratyphi A ($14.1{\pm}1.1mm$), E. aerogenes ($5.6{\pm}0.4mm$) and S. marcescens ($14.25{\pm}0.2mm$). GC-MS analysis revealed the dominant presence of oleic acid C 18.1 (63.18%), n-hexadecanoic acid (6.17%), and ethyl oleate (4.93%) and potent molecules such as 8-[(2E)-2-(3-hydroxybenzylidene)hydrazinyl]-1,3,7-trimethyl-3,7-dihydro-1H-purine-2,6-dione, 2-(Dimethylamino)ethyl (1Z)-N-hydroxy-2-(4-morpholinyl)-2-oxoethanimidothioate, Fluorene in the F41, and virtual study revealed that these molecules are likely responsible for the antibacterial activities of F41. Hence, further investigation deserves on purification and characterization of the active metabolites from T. atroviride strain KNUP001 towards developing molecular leads to effective antibacterial drugs, and non-toxic to host cells.

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