• Title/Summary/Keyword: Molecular Surface

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Vimentin Is Involved in Peptidylarginine Deiminase 2-Induced Apoptosis of Activated Jurkat Cells

  • Hsu, Pei-Chen;Liao, Ya-Fan;Lin, Chin-Li;Lin, Wen-Hao;Liu, Guang-Yaw;Hung, Hui-Chih
    • Molecules and Cells
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    • v.37 no.5
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    • pp.426-434
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    • 2014
  • Peptidylarginine deiminase type 2 (PADI2) deiminates (or citrullinates) arginine residues in protein to citrulline residues in a $Ca^{2+}$-dependent manner, and is found in lymphocytes and macrophages. Vimentin is an intermediate filament protein and a well-known substrate of PADI2. Citrullinated vimentin is found in ionomycin-induced macrophage apoptosis. Citrullinated vimentin is the target of anti-Sa antibodies, which are specific to rheumatoid arthritis, and play a critical role in the pathogenesis of the disease. To investigate the role of PADI2 in apoptosis, we generated a Jurkat cell line that overexpressed the PADI2 transgene from a tetracycline-inducible promoter, and used a combination of 12-O-tetradecanoylphorbol-13-acetate and ionomycin to activate Jurkat cells. We found that PADI2 overexpression reduced the cell viability of activated Jurkat cells in1a dose- and time-dependent manner. The PADI2-overexpressed and -activated Jurkat cells presented typical manifestations of apoptosis, and exhibited greater levels of citrullinated proteins, including citrullinated vimentin. Vimentin overexpression rescued a portion of the cells from apoptosis. In conclusion, PADI2 overexpression induces apoptosis in activated Jurkat cells. Vimentin is involved in PADI2-induced apoptosis. Moreover, PADI2-overexpressed Jurkat cells secreted greater levels of vimentin after activation, and expressed more vimentin on their cell surfaces when undergoing apoptosis. Through artificially highlighting PADI2 and vimentin, we demonstrated that PADI2 and vimentin participate in the apoptotic mechanisms of activated T lymphocytes. The secretion and surface expression of vimentin are possible ways of autoantigen presentation to the immune system.

Adipose-Derived Stem Cells Suppress Inflammation Induced by IL-1β through Down-Regulation of P2X7R Mediated by miR-373 in Chondrocytes of Osteoarthritis

  • Jin, Rilong;Shen, Miaoda;Yu, Liedao;Wang, Xuanwei;Lin, Xiangjin
    • Molecules and Cells
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    • v.40 no.3
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    • pp.222-229
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    • 2017
  • Adipose-derived stem cells (ADSCs) were previously considered to have an anti-inflammatory effect, and Interleukin-$1{\beta}$ ($IL-1{\beta}$) was found to be a pro-inflammatory factor in chondrocytes, but the mechanism underlying ADSCs and $IL-1{\beta}$ is unclear. In this study, we investigate whether P2X7 receptor (P2X7R) signalling, regulated by microRNA 373 (miR-373), was involved in the ADSCs and $IL-1{\beta}$ mediated inflammation in osteoarthritis (OA). Chondrocytes were collected from 20 OA patients and 20 control participants, and ADSCs were collected from patients who had undergone abdominal surgery. The typical surface molecules of ASDCs were detected by flow cytometry. The level of nitric oxide (NO) was determined by Griess reagent. Concentrations of prostaglandin E2 (PGE2), interleukin 6 (IL-6), matrix metallopeptidase 3 (MMP-3) were detected by enzyme-linked immunosorbent assay (ELISA). The expressions of IL-6, MMP-3, miR-373 and P2X7R were determined by real-time polymerase chain reaction (PCR), and Western blot was used to detect the protein expression of P2X7R. The typical potential characters of ADSCs were verified. In chondrocytes or OA tissues, the miR-373 expression level was decreased, but the P2X7R expression was increased. $IL-1{\beta}$ stimulation increased the level of inflammatory factors in OA chondrocytes, and ADSCs co-cultured with $IL-1{\beta}$-stimulated chondrocytes decreased the inflammation. OA chondrocytes transfected with the miR-373 inhibitor increased the inflammation level. The miR-373 mimic suppressed the inflammation by targeting P2X7R and regulated its expression, while its effect was reversed by overexpression of P2X7R. $IL-1{\beta}$ induced inflammation in OA chondrocytes, while ADSCs seemed to inhibit the expression of P2X7R that was regulated by miR-373 and involved in the anti-inflammatory process in OA.

Characterization of Streptococcosis Occurrence and Molecular Identification of the Pathogens of Cultured Flounder in Jeju Island (제주지역 양식넙치의 연쇄구균증 발생동향 및 원인균에 대한 분자적 동정)

  • Jeong, Yong-Uk;Kang, Chul-Young;Kim, Min-Ju;Heo, Moon-Soo;Oh, Duck-Chul;Kang, Bong-Jo
    • Korean Journal of Microbiology
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    • v.42 no.3
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    • pp.199-204
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    • 2006
  • Streptococcosis of olive flounder(Paralichthys olivaceus) is an important bacterial disease in Jeju island. In this study, we investigated monthly infection pattern of this disease in different size of the flounder fish. Even though the disease occurred throughout the year, the infection ratio was relatively higher in the months with warm water season. The infection was more prevalent in adult flounder over 30 cm total length compare to these of small size fish. Two infectious species of streptococcosis pathogens were detected by multiplex PCR assay. Detection ratios of Streptococcus iniae and S. parauberis reached up to 46% and 54%, respectively, from June 2003 to May 2005 in Jeju island. S. iniae occurred intensively from September to October, whereas S. parauberis reported from March to May. S. iniae and S. parauberis infections of cultured flounder share some common features, but clinical findings showed considerable differences between two diseases. Distended abdomen, protruded anus and ascitic fluid in the peritoneal cavity are evident lesions detected in S. iniae infection, whereas, flounders infected by S. parauberis showed prominent lesions such as darkened surface and haemorrhaging in the non-ocular side. Both streptococcosis pathogens were sensitive to antibiotics, such as ampicillin and amoxicillin. However, S. iniae strains were more sensitive to doxycycline, erythromycin and oxytetracycline than S. parauberis strains.

Geochemistry of Mn Scales Formed in Groundwater in the Damyang Area (담양 지역 음용 지하수에 형성된 망간 스케일에 대한 지구화학)

  • Park, Cheon-Young;Kim, Seoung-Ku;Shin, In-Hyun;Ahan, Kun-Sang;Kim, Young-In
    • Journal of the Korean earth science society
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    • v.27 no.3
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    • pp.313-327
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    • 2006
  • This study investigated the geochernical characteristics of Mn scale formed in groundwater wells at the Damyang area. The composition of Mn scale consists mainly of MnO and $SiO_2$. The content of Mn ranges from56.61wt.% to 68.69wt.%, and $SiO_2$ content ranges from 1.56wt.% to 10.45wt.%. The contents of Mo and Ba in Mn scale increased with increased depth; whereas, the content of Zn and Pb decreased with increased depth. Birnessite, quartz and feldspars were identified in Mn scales using x-ray powder diffraction studies. The IR absorption bands for Mn scales show major absorption band due to OH stretching, adsorbed molecular water, and birnessite stretching, respectively. In the SEM and EDS analysis, the Mn scale consists of botryoidal, spherical, spherulite, and empty straw structure. Those structure may be precipitated simply due to oversaturation with concentrated Mn content or may be formed through biogenic precipitation by Lepthothrix discophora. Under microanalysis using EDS on those structure surface of Mn scales, the Mn atomic percent range from 28 to 44, and such elements revealed the presence of Si, K, Na, Ca, Cl, Cu, Zn, and Ba.

Reactive Ceramic Membrane Incorporated with Iron Oxide Nanoparticle for Fouling Control (산화철 나노입자 부착 반응성 세라믹 멤브레인의 막 오염 제어)

  • Park, Hosik;Choi, Heechul
    • Journal of Korean Society of Environmental Engineers
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    • v.35 no.2
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    • pp.144-150
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    • 2013
  • Hybrid ceramic membrane (HCM) processes that combined ozonation with a ceramic membrane (CM) or a reactive ceramic membrane (RM), an iron oxide nanoparticles (IONs) incorporated-CM were investigated for membrane fouling control. Alumina disc type microfiltration and ultrafiltration membranes doped with IONs by sintering method were tested under varying mass fraction of IONs. Scanning electron microscope (SEM) images showed that IONs were well-doped on the CM surface and doped IONs were approximately 50 nm in size. Change in the pure water permeability of RM was negligible compared to that of CM. These results indicate that IONs incorporation onto CM had little effect on CM performance in terms of the flux. Natural organic matter (NOM) fouling and fouling recovery patterns during HCM processes confirmed that the RM-ozonation process enhanced the destruction of NOM and reduced the extent of fouling more than the CM-ozonation process by hydroxyl radical formation in the presence of IONs on RM. In addition, analyses of NOM in the feed water and the permeate showed that the efficiency of membrane fouling control results from the NOM degradation during HCM processes; leading to removal and transformation of relatively high contents of aromatic, high molecular weight and hydrophobic NOM fractions.

Saprolegnia parasitica Isolated from Rainbow Trout in Korea: Characterization, Anti-Saprolegnia Activity and Host Pathogen Interaction in Zebrafish Disease Model

  • Shin, Sangyeop;Kulatunga, D.C.M.;Dananjaya, S.H.S.;Nikapitiya, Chamilani;Lee, Jehee;De Zoysa, Mahanama
    • Mycobiology
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    • v.45 no.4
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    • pp.297-311
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    • 2017
  • Saprolegniasis is one of the most devastating oomycete diseases in freshwater fish which is caused by species in the genus Saprolegnia including Saprolegnia parasitica. In this study, we isolated the strain of S. parasitica from diseased rainbow trout in Korea. Morphological and molecular based identification confirmed that isolated oomycete belongs to the member of S. parasitica, supported by its typical features including cotton-like mycelium, zoospores and phylogenetic analysis with internal transcribed spacer region. Pathogenicity of isolated S. parasitica was developed in embryo, juvenile, and adult zebrafish as a disease model. Host-pathogen interaction in adult zebrafish was investigated at transcriptional level. Upon infection with S. parasitica, pathogen/antigen recognition and signaling (TLR2, TLR4b, TLR5b, NOD1, and major histocompatibility complex class I), pro/anti-inflammatory cytokines (interleukin $[IL]-1{\beta}$, tumor necrosis factor ${\alpha}$, IL-6, IL-8, interferon ${\gamma}$, IL-12, and IL-10), matrix metalloproteinase (MMP9 and MMP13), cell surface molecules ($CD8^+$ and $CD4^+$) and antioxidant enzymes (superoxide dismutase, catalase) related genes were differentially modulated at 3- and 12-hr post infection. As an anti-Saprolegnia agent, plant based lawsone was applied to investigate on the susceptibility of S. parasitica showing the minimum inhibitory concentration and percentage inhibition of radial growth as $200{\mu}g/mL$ and 31.8%, respectively. Moreover, natural lawsone changed the membrane permeability of S. parasitica mycelium and caused irreversible damage and disintegration to the cellular membranes of S. parasitica. Transcriptional responses of the genes of S. parasitica mycelium exposed to lawsone were altered, indicating that lawsone could be a potential anti-S. parasitica agent for controlling S. parasitica infection.

Preparation of Amino Acid Copolymers/water-insoluble Drug Nanoparticles: Polymer Properties and Processing Variables (아미노산 공중합체/난용성 약물 나노입자의 제조: 고분자 특성 및 가공변수)

  • Yoo Ji Youn;Lee Soo-Jeong;Ahn Cheol-Hee;Choi Ji-Yeun;Lee Jonghwi
    • Polymer(Korea)
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    • v.29 no.5
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    • pp.440-444
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    • 2005
  • An increase in the surface area of drugs by reducing particle sizes from microns to nanometers has been known as an efficient method to improve the bioavailability of water-insoluble drugs. To prevent drug nanoparticles from aggregation during the processes of drug formulation, a limited number of pharmaceutical inactive ingredients such as hydroxypropyl cellulose has been employed as stabilizers or dispersants. In this study, copolymers of hydrophilic and hydrophobic amino acids were synthesized by the ring opening polymerization of their N-carboxyanhydride monomers and evaluated as novel candidates to stabilize the nanoparticles of a water insoluble drug, naproxen. Naproxen nanoparticles stabilized by synthesized amino acid copolymers were successfully prepared in the size of $200\~500nm$ in 60 min by a wet comminution process. Particle size analysis showed that the effective stabilization performance of copolymers required the hydrophobic moiety content to be higher than $10 mol\%$. However, the molecular weight and morphology of copolymers was not the critical parameters in determining the particle size reduction. Their particle size was found to be stable up to 14 days without significant aggregation.

Synthesis and Characterization of Heat Resistant Organophilic Layered Silicate Modified with Oligo(amic acid)s Having Alkyl Side Chains and Their Nanocomposites (알킬기가 도입된 올리고 아믹산 구조를 가진 고내열 친유기 층상 실리케이트의 제조 및 이를 이용한 나노복합재의 특성평가)

  • Han Ji Yun;Won Jong Chan;Lee Jae Heung;Suh Kyung-Do;Kim Yong Seok
    • Polymer(Korea)
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    • v.29 no.5
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    • pp.451-456
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    • 2005
  • In the field of designing of nano-fillers of polyimide nanocomposites, the two strategic points are the heat-resistance and compatibility with polyimide, a matrix polymer. In this study, we designed oligo(amic acid) having alkyl side chains and terminal amine groups to satisfy previous requirements and studied the modification of surface of layered silicates. Oligo(amic acid)s were prepared by the reaction of diamine monomers and PMDA and their molecular weight was controlled in about 2000g/mol. After that, acidification and ion exchange reaction led to the high-temperature organophilic layered silicate (OLS). XRD pauerns of OLS showed the more increased gallery spacing by $4{\AA}$ than that of the pristine layered silicate and the initial decomposition temperatures of OLS were in above $280^{\circ}C$. The polyimide nanocomposite films based on heat resistant OLS showed that the OLSs were well dispersed through the matrix and their CTEs showed a decrease of $26\%$ compared with pristine polyimide films.

Characteristics of BCNU-loaded PLGA Wafers (BCNU를 함유한 생분해성 PLGA 웨이퍼의 특성분석)

  • 안태군;강희정;이진수;성하수;정제교
    • Polymer(Korea)
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    • v.26 no.5
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    • pp.691-700
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    • 2002
  • Interstitial therapy using biodegradable polymeric device loaded with anticancer agent can deliver the drug to the tumor site at high concentration, resulting in an increase of therapeutic efficacy. 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU, carmustine) is most commonly used as chemotherapeutic agent for brain tumors. The design of implantable device is regarded as an important factor lot the efficient delivery of antitumor agent to targeting site. In order to control the release profile of drug, the release pattern of BCNU with the changes of various dimension and additives was investigated. The PLGA wafers containing 3.85, 10, 20 and 30% of BCNU were prepared in various shape (diameter of 3, 5 and 10 mm, thickness of 0.5, 1 and 2 mm) by direct compression method. In vitro drug release profile of BCNU-loaded PLGA wafers could be controlled by changing the dimension of wafers such as initial drug content, weight, diameter, thickness, volume and surface area of wafers, as well as PLGA molecular weight and additives. Drug release from BCNU-loaded PLGA wafers was facilitated with an increase of BCNU-loading amount or presence of poly(N-vinylpyrrolidone)(PVP) or sodium chloride (NaCl). The effects of various geometric factors and additives on the BCNU release pattern were confirmed by the investigation of mass loss and morphology of BCNU-loaded PLGA wafers.

Effect of Gamitonggyu-tang on Secretion of Airway Mucin and Contractility of Tracheal Smooth Muscle (가미통규탕(加味通竅湯)이 호흡기 뮤신 분비 및 기관 평활근 긴장도에 미치는 영향)

  • Lee, Nam-Yeol;Han, Jae-Kyung;Kim, Yun-Hee
    • The Journal of Pediatrics of Korean Medicine
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    • v.21 no.3
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    • pp.109-124
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    • 2007
  • Objectives In the present study, the author intended to investigate whether Gamitonggyu-tang (GTT) significantly affects (since the subject is GTT, you need an 's') in vivo and in vitro mucin secretion from airway epithelial cells. Methods In vivo experiment, mice's mucin which is on a hypersecretion of an airway, mice's tracheal goblet cells in hyperplasia and mice's intraepithelial mucosubstances were exposed with SO2 for 3 weeks. Effects of orally-administered GTT for 1 week on in vivo mucin secretion and hyperplasia of tracheal goblet cells were assessed by using enzyme-linked immunosorbent assay (ELISA) and staining goblet cells with alcian blue. In vitro experiment, confluent hamster tracheal surface epithelial (HTSE) cells were metabolically radiolabeled with 3H-glucosamine for 24 hrs and chased for 30 min in the presence of GTT to figure out the effectiveness of 3H-mucin secretion. Total elution profiles of control spent media and treatment sample through Sepharose CL-4B column were analyzed.Possible cytotoxicities of each agent were assessed by measuring lactate dehydrogenase (LDH) release. Also, the effect of GTT on contractility of isolated tracheal smooth muscle was investigated. Results (1) GTT inhibited hypersecretion of in vivo mucin. However, it did not affect the increase the number of goblet cells (2) GTT significantly increased mucin release from cultured HTSE cells, without significant cytotoxicity (3) GTT chiefly affected the 'mucin' secretion and did not affect the secretion of the other releasable glycoproteins with less molecular weight than mucin (4) GTT did not affect Ach-induced contraction of isolated tracheal smooth muscle.Conclusions This result suggests that GTT can increase mucin secretion during short-term treatment (in vitro) whereas it can inihibit hypersecretion of mucin during long-term treatment (in vivo). The author suggests that the effect GTT with their components should be further investigated and it is valuable to find from oriental medical prescriptions, novel agents which might regulate mucin secretion from airway epithelial cells.

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