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Expression of Epstein-Barr Virus Gene and Clonality of Infiltrated T Lymphocytes in Epstein-Barr Virus-associated Gastric Carcinoma

  • Lee, Jae-Myun;Kim, Ho-Guen;Noh, Sung-Hoon;Lee, Won-Young;Kim, Se-Jong;Park, Jeon-Han
    • IMMUNE NETWORK
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    • v.11 no.1
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    • pp.50-58
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    • 2011
  • Background: Epstein-Barr virus associated gastric lymphoepithelioma-like carcinoma (LELC) is characterized by the intensive infiltration of lymphoid cells, the presence of EBV, and the better prognosis over typical adenocarcinoma. Thus, it was assumable that viral latent proteins may be responsible for the recruitment of a certain T cell repertoire to EBV-associated gastric carcinoma. Methods: To examine above possibility, EBV gene expression in gastric carcinoma tissues and usage of TCR among the tumor infiltrating lymphocytes were analyzed. Results: EBV specific DNA and EBERs RNA were detected in 4 out of 30 patients. RT-PCR analysis revealed that all 4 of EBV-positive tumor tissues expressed EBNA1 mRNA and BARTs and LMP2a was detected only one sample out of 4. However, the EBNA2 and LMP-1 transcripts were not detected in these tissues. $CD8^+$ T cells were the predominant population of infiltrating lymphocytes in the EBV-positive gastric carcinoma. According to spectra type analysis of infiltrating T cells, 10 predominant bands were detected by TCR $V{\beta}$ CDR3 specific RT-PCR from 4 EBV-positive tumor tissues. Sequence analysis of these bands revealed oligoclonal expansion of T cells. Conclusion: These findings suggest that clonally expanded T cells in vivo might be a population of cytotoxic T cells reactive to EBV-associated gastric carcinoma.

Microstructure and Corrosion Resistance of Ti-15Sn-4Nb Alloy with Hf Adding Element (Hf가 첨가된 생체용 Ti-15Sn-4Nb 합금의 미세조직 및 내식성)

  • Lee, Doh-Jae;Lee, Kyung-Ku;Cho, Kyu-Zong;Yoon, Taek-Rim;Park, Hyo-Byung
    • Journal of Technologic Dentistry
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    • v.23 no.1
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    • pp.55-64
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    • 2001
  • This study is focusing on the improvement of problems of Ti-6Al-4V alloy. A new Ti based alloy, Ti-15Sn-4Nb, have designed to examine any possibility of improving the mechanical properties and biocompatibility. Specimens of Ti alloys were melted in vacuum arc furnace and homogenized at $100^{\circ}C$ for 24h. All specimens were solution treated at $812^{\circ}C$ and aged at $500^{\circ}C$ for 10h. The corrosion resistance of Ti alloys was evaluated by potentiodynamic polarization test and immersion test inl%Lactic acid solutions. Ti-15Sn-4Nb system alloys showed Widmanstatten microstructure after solution treatment which is typical microstructure of ${\alpha}+{\beta}$ type Ti alloys. Analysing the corrosion resistance of Ti alloys, it was concluded that the passive films of Ti-15Sn-4Nb system alloys are more stable than that of Ti-6Al-4V alloys. Also, the corrosion resistance of Ti-15Sn-4Nb system alloys was improved with adding elements, Hf. It was analysed that the passive film of the Ti-15Sn-4Nb alloy which was formed in air atmosphere was consisted of TiO2, SnO and NbO through X-ray photoelectron spectroscopy(XPS) analysis.

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Influence of Reproductive Status, Serum Type and Estradiol-17β Supplementation on the in vitro Maturation of Canine Oocytes

  • Heru, Fibrianto Yuda;Kim, Min-Kyu
    • Korean Journal of Agricultural Science
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    • v.35 no.2
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    • pp.167-176
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    • 2008
  • Supplementation of serum and estrogen in in vitro maturation(IVM) medium was shown to improve embryo development and quality in several species. This study investigates the effect of ovarian estrus stage on canine oocyte quality and supplementation of medium with canine serum or estrogen on IVM of canine oocytes. As results, in experimental 1, IVM oocytes collected from follicular stage ovaries to MII stages($10.2{\pm}1.5%$) was higher (p<0.05) with 10% canine estrus stage serum than control($1.3{\pm}1.6%$), anoestrus stage serum($4.0{\pm}1.6%$), luteal stage serum($2.7{\pm}1.7%$) and 10% FBS($1.3{\pm}1.6$). In experimental 2, 10% canine estrus stage serum supplementation has highest maturation rate to MII stages($10.0{\pm}1.8%$) and there were significant differences(P<0.05) with another treatment in follicular stages group. In order to investigate the synergic effect of estrous serum and estrogen supplementation, different estrous stage groups of oocytes were cultured with 2 ug/ml estrogen plus various concentrations of different reproductive stage serum and FBS(experimental 3). As results, the rate of maturation to metaphase II(MII) stage was significantly higher(p<0.05) in oocytes from the follicular stage supplemented with estrogen and 10% canine estrus stage serum(11.5%) compared to the other groups(6.0 - 8.8%). The present study was demonstrated that canine serum and the estrus cycle of the bitch affect the meiotic competence of oocytes. Hormonal influences within the follicle may be one of the factors responsible for the greater proportion of maturation of oocyte to MII from bitches at the follicular phase.

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Mechanism on the Hydrolysis of Cinnamonitrile in Strong Acid (强酸性 溶液中에서 Cinnamonitrile의 加水分解 反應메카니즘)

  • Ki-Sung Kwon;Nack-Do Sung;Tae-Rin Kim;Jeon, Yong Gu
    • Journal of the Korean Chemical Society
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    • v.28 no.6
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    • pp.418-424
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    • 1984
  • Rate constants for the hydrolysis of cinnamonitrile in the concentration range of 1 ∼ 5M of perchloric acid at 25$^{\circ}$C have been determined by UV spectrophotometry and from the Bunnett equations, hydration parameters (${\omega}$ = 9.8, ${\omega}^*$ = 0.42 & ${\phi}$=1.6) were obtained. CNDO/2 MO calculations were performed to determine relative stability, net charges, and overlap population of various conformational isomers. The results show that the (E)-planar is more stable than the (Z)-planar and protonation is favored on the nitrogen atom. On the basis of above findings, the acid hydrolysis is initiated by the protonation of the nitrogen atom of cinnamonitrile and then water molecule acting as nucleophile and as a proton transfer agent in the rate determining step. In the transition state of the acid hydrolysis, nucleophilic addition of water molecule occurs by sigma approach to the positively charged $C_7({\alpha}$) atom of the conjugate acid. As the results, we may conclude that the hydrolysis of cinnamonitrile in the strong acidic media proceeds through the A-2 type mechanism.

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The Arabidopsis Phytocystatin AtCYS5 Enhances Seed Germination and Seedling Growth under Heat Stress Conditions

  • Song, Chieun;Kim, Taeyoon;Chung, Woo Sik;Lim, Chae Oh
    • Molecules and Cells
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    • v.40 no.8
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    • pp.577-586
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    • 2017
  • Phytocystatins (PhyCYSs) are plant-specific proteinaceous inhibitors that are implicated in protein turnover and stress responses. Here, we characterized a PhyCYS from Arabidopsis thaliana, which was designated AtCYS5. RT-qPCR analysis showed that the expression of AtCYS5 in germinating seeds was induced by heat stress (HS) and exogenous abscisic acid (ABA) treatment. Analysis of the expression of the ${\beta}-glucuronidase$ reporter gene under the control of the AtCYS5 promoter showed that AtCYS5 expression during seed germination was induced by HS and ABA. Constitutive overexpression of AtCYS5 driven by the cauliflower mosaic virus 35S promoter led to enhanced HS tolerance in transgenic Arabidopsis, which was characterized by higher fresh weight and root length compared to wild-type (WT) and knockout (cys5) plants grown under HS conditions. The HS tolerance of AtCYS5-overexpressing transgenic plants was associated with increased insensitivity to exogenous ABA during both seed germination and post-germination compared to WT and cys5. Although no HS elements were identified in the 5'-flanking region of AtCYS5, canonical ABA-responsive elements (ABREs) were detected. AtCYS5 was upregulated in ABAtreated protoplasts transiently co-expressing this gene and genes encoding bZIP ABRE-binding factors (ABFs and AREB3). In the absence of ABA, ABF1 and ABF3 directly bound to the ABREs in the AtCYS5 promoter, which activated the transcription of this gene in the presence of ABA. These results suggest that an ABA-dependent pathway plays a positive role in the HS-responsive expression of AtCYS5 during seed germination and post-germination growth.

Flagellin-Stimulated Production of Interferon-β Promotes Anti-Flagellin IgG2c and IgA Responses

  • Kang, Wondae;Park, Areum;Huh, Ji-Won;You, Gihoon;Jung, Da-Jung;Song, Manki;Lee, Heung Kyu;Kim, You-Me
    • Molecules and Cells
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    • v.43 no.3
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    • pp.251-263
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    • 2020
  • Flagellin, a major structural protein of the flagellum found in all motile bacteria, activates the TLR5- or NLRC4 inflammasome-dependent signaling pathway to induce innate immune responses. Flagellin can also serve as a specific antigen for the adaptive immune system and stimulate anti-flagellin antibody responses. Failure to recognize commensal-derived flagellin in TLR5-deficient mice leads to the reduction in anti-flagellin IgA antibodies at steady state and causes microbial dysbiosis and mucosal barrier breach by flagellated bacteria to promote chronic intestinal inflammation. Despite the important role of anti-flagellin antibodies in maintaining the intestinal homeostasis, regulatory mechanisms underlying the flagellin-specific antibody responses are not well understood. In this study, we show that flagellin induces interferon-β (IFN-β) production and subsequently activates type I IFN receptor signaling in a TLR5- and MyD88-dependent manner in vitro and in vivo. Internalization of TLR5 from the plasma membrane to the acidic environment of endolysosomes was required for the production of IFN-β, but not for other pro-inflammatory cytokines. In addition, we found that anti-flagellin IgG2c and IgA responses were severely impaired in interferon-alpha receptor 1 (IFNAR1)-deficient mice, suggesting that IFN-β produced by the flagellin stimulation regulates anti-flagellin antibody class switching. Our findings shed a new light on the regulation of flagellin-mediated immune activation and may help find new strategies to promote the intestinal health and develop mucosal vaccines.

Nano-Structure Control of SiC Hollow Fiber Prepared from Polycarbosilane (폴리카보실란으로부터 제조된 탄화규소 중공사의 미세구조제어)

  • Shin, Dong-Geun;Kong, Eun-Bae;Cho, Kwang-Youn;Kwon, Woo-Tek;Kim, Younghee;Kim, Soo-Ryong;Hong, Jun-Sung;Riu, Doh-Hyung
    • Journal of the Korean Ceramic Society
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    • v.50 no.4
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    • pp.301-307
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    • 2013
  • SiC hollow fiber was fabricated by curing, dissolution and sintering of Al-PCS fiber, which was melt spun the polyaluminocarbosilane. Al-PCS fiber was thermally oxidized and dissolved in toluene to remove the unoxidized area, the core of the cured fiber. The wall thickness ($t_{wall}$) of Al-PCS fiber was monotonically increased with an increasing oxidation curing time. The Al-PCS hollow fiber was heat-treated at the temperature between 1200 and $2000^{\circ}C$ to make a SiC hollow fibers having porous structure on the fiber wall. The pore size of the fiber wall was increased with the sintering temperature due to the decomposition of the amorphous $SiC_xO_y$ matrix and the growth of ${\beta}$-SiC in the matrix. At $1400^{\circ}C$, a nano porous wall with a high specific surface area was obtained. However, nano pores grew with the grain growth after the thermal decomposition of the amorphous matrix. This type of SiC hollow fibers are expected to be used as a substrate for a gas separation membrane.

Substrate reduction therapy in three patients with Gaucher disease

  • Kim, Soo Hyun;Kang, Eungu;Kim, Yoon-Myung;Kim, Gu-Hwan;Choi, In-Hee;Choi, Jin-Ho;Yoo, Han-Wook;Lee, Beom Hee
    • Journal of Genetic Medicine
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    • v.13 no.2
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    • pp.72-77
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    • 2016
  • Purpose: Gaucher disease (GD) is the most common lysosomal storage disease caused by beta-glucocerebrosidase (GBA) deficiency. Oral substrate reduction therapy with miglustat ($Zavesca^{(R)}$) was approved for the treatment of adults with GD type 1, for whom enzyme replacement therapy (ERT) is unsuitable or not a therapeutic option. In this study, we report the effect of miglustat ($Zavesca^{(R)}$) in three Korean GD patients. Materials and Methods: Clinical findings comprising age at diagnosis, presenting signs, laboratory findings at diagnosis, GBA activity and mutations, and clinical courses of the three patients were reviewed. Results: Miglustat was administered to three patients who reported allergic reactions during intravenous imiglucerase infusions. One patient withdrew after 15 months of miglustat administration owing to continuous elevation of disease biomarker levels (chitotriosidase, acid phosphatase, and angiotensin-converting enzyme). Poor adherence to medication was suspected but was denied by the patient. In the other two patients, platelet count and levels of hemoglobin and other biomarkers remained stable during miglustat administration. However, they suffered from severe diarrhea and weight loss, which led to miglustat discontinuation after 1 and 12 months of administration. Conclusion: Our study shows that although miglustat is suggested to GD patients as an alternative treatment to ERT, significant adverse reactions may lead to discontinuation of miglustat. In addition, it is difficult to monitor the drug adherence.

Skin Permeation and Anti-inflammatory Effects of Hydrolyzed Products of Gardeniae Fructus Extracts (치자 엑스의 가수분해에 의한 피부투과 및 항염효과)

  • Yang, Jae-Heon;Lee, Nam-Hee
    • Journal of Pharmaceutical Investigation
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    • v.34 no.2
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    • pp.115-123
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    • 2004
  • Geniposide and its related iridoid compounds have been used in traditional herbal medicine for thε treatment of Jaundice hepatic diseases and various inflammatorys. For the purpose to increase trandsdermal absorption, the hydrolyzed products of Gardeniae Furctus were identified and assayed of active ingredients and investigated trandsdermal absorption and anti-inflammatory effects. Geniposide was hydrolyzed to genipin by ${\beta}-glucosidase$ and it was suggested that genipin was more suitable form than geniposide for transdermal absorption by its lipophilic property. Using Franz type diffusion cell and the skin of hairless mouse, the permeation rate of hydrolyzed products and their emulsion preparation were determined. Genipin have more increased absorption ratio through the skin of hairless mouse than geniposide. Also, the emulsion of hydrolyzed products of extracts showed higher permeability than that of nonhydrolyzed preparations. After 9 hours $280.85\;{\mu}g/cm^2$ of genipin was absorbed and $193.52\;{\mu}g/cm^2$ in case of geniposide. The Js of geniposide and genipin were $26.27{\pm}4.11\;{\mu}g/cm^2/hr$ and $40.35{\pm}5.04\;{\mu}g/cm^2/hr$ respectively. After carrageenan injection, the swelling was increased repidly to 24 hr and maintained as plateau. but emulsion group weer reached about 2.5 mL and the swelling decreased successively form 24 hr to 72 hr. The anti-inflammation effects of extracts and hydrolyzed products emulsion were increased with significant difference with control group after 24 hr, 48 hr and 72 hr. In carrageenan induced edema, inhibition of swelling was increased in case of hydrolyzed product emulsion compare with nonhydrolyzed group at 24 hr, 48 hr and 72 hr after swelling. In histological study, the anti-inflammatory effects of hydrolyzed products were remarkable at 48 hr and 72 hr compare with nonhydrolyzed. Hydrolyzed products of Gardeniae Fructus extracts containing genipin would be a suitable preparation to increase the transdermal absorption and anti-inflammatory effects.

Formulation of Caffeine Nasal Sprays and Its Enhanced Permeation through Rabbit Nasal Mucosa (카페인의 비강 분무액의 제제설계 및 점막 투과 증진)

  • Noh, Eun-Sun;Chun, In-Koo
    • Journal of Pharmaceutical Investigation
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    • v.34 no.2
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    • pp.131-138
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    • 2004
  • This study was aimed to investigate the feasibility of nasal delivery of caffeine for the elimination of sleepiness. The effects of various vehicles, solubilizers, and enhancers on the permeation of caffeine through rabbit nasal mucosa was observed. The permeation study was carried out using a Franz-type permeation system at $37^{\circ}C$, and the amount of caffeine permeated though the rabbit nasal mucosa was determined by a validated HPLC. The apparent solubility and phys iochemical stability of caffeine in various nasal formulations were was determined. The effect of hydrotropes and modified cyclodextrins on the solubility of caffeine in water was determined by equilibrium solubility method. The solubility of caffeine in water was 29 mg/mL at $30^{\circ}C$. The addition of sodium benzoate and nicotinamide at 10% improved the solubility of caffeine (115 and 132 mg/mL, respectively) in aqueous solution. The flux of caffeine though the nasal mucosa from aqueous solution was $2.1{\pm}0.26\;mg/cm^2/hr$. The addition of sodium benzoate reduced its permeation $(1.4{\pm}0.01\;mg/cm^2/hr)$, but sodium benzoate with 5% $2HP{\beta}CD$ and 0.03% monoterpenes increased its permeation $(2.4{\pm}0.04\;mg/cm^2/hr)$ markedly. The addition of nicotinamide also increased also increased its permeation $(2.5{\pm}0.36\;mg/cm^2/hr)$. markedly. As the concentration of caffeine in nasal formulation increased, the permeation flux increased linearly. Caffeine was stable physicochemically and enzymatically in the nasal mucosa extract at $37^{\circ}C$. These results suggest that caffeine can be efficiently delivered nasally and the development of nasal formulation will be feasible.