• 제목/요약/키워드: EDTA treatment

검색결과 237건 처리시간 0.027초

($CO_2$)레이저 치료에 의한 치근표면의 구조적 변화 (Strutural Change of Root Surface Following $CO_2$ Laser Treatment)

  • 김도영;한수부;고재승;박성현;계승범;황광세;김우성
    • Journal of Periodontal and Implant Science
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    • 제27권3호
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    • pp.469-478
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    • 1997
  • The purpose of this study was to evaluate the structural change of root surface and the occlusion of dentinal tubule following $CO_2$ laser treatment. Seven extracted healthy human premolar werw curetted, sectioned, and four specimens were randomly assigned to each of 6 different treatment groups : 1) untreated EDTA etched control: 2) root plande only: 3) $CO_2$ laser treated with 2W mode 6(10msec/pulse, 20pps) for 1 minute: 4) $CO_2$ laser treated with 2W mode 6(lOmsec/pulse, 20pps) for 2 minutes: 5) $CO_2$ laser treated with 2W mode 7(20msec/pulse, 20pps) for 1 minute: 6) $CO_2$ laser treated with 2W mode 7(20msec/pulse, 20pps) for 2 minutes. Following the prescribed treatment, the specimens were prepared for SEM evaluation. Results showed that $CO_2$ laser may be effective to occlude dentinal tubules tor dentin sensitivity treatment. The effect of dentinal tubule occlusion was enhanced with increasing the total energy level lased to specimen regardless of lasing mode. The structural changes of root surfaces were restricted to superficies, and these changes included fissuring, charring, crater formation over the smooth lava like texture. The charring and crater formation implying root damage was observed in the case of the longer duration of a pulse. The results of the present study suggests that the pulsed $CO_2$ laser with shorter pulse duration and longer exposure time can be used effectively in order to obtain the optimal dentinal tubule occlusion with minimal root damage.

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Validation of Methods for Isolation and Culture of Alpaca Melanocytes: A Novel Tool for In vitro Studies of Mechanisms Controlling Coat Color

  • Bai, Rui;Sen, Aritro;Yu, Zhihui;Yang, Gang;Wang, Haidong;Fan, Ruiwen;Lv, Lihua;Lee, Kyung-Bon;Smith, George W;Dong, Changsheng
    • Asian-Australasian Journal of Animal Sciences
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    • 제23권4호
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    • pp.430-436
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    • 2010
  • The objective of the present studies was to develop and validate a system for isolation, purification and extended culture of pigment-producing cells in alpaca skin (melanocytes) responsible for coat color and to determine the effect of alpha melanocyte stimulating hormone treatment on mRNA expression for the melanocortin 1 receptor, a key gene involved in coat color regulation in other species. Skin punch biopsies were harvested from the dorsal region of 1-3 yr old alpacas and three different enzyme digestion methods were evaluated for effects on yield of viable cells and attachment in vitro. Greatest cell yields and attachment were obtained following dispersion with dispase II relative to trypsin and trypsin-EDTA treatment. Culture of cells in medium supplemented with basic fibroblast growth factor, bovine pituitary extract, hydrocortisone, insulin, 12-O-tetradecanolphorbol-13-acetate and cholera toxin yielded highly pure populations of melanocytes by passage 3 as confirmed by detection of tyrosinase activity and immunocytochemical localization of melanocyte markers including tyrosinase, S-100 and micropthalmia-associated transcription factor. Abundance of mRNA for tyrosinase, a key enzyme in melanocyte pigment production, was maintained through 10 passages showing preservation of melanocyte phenotypic characteristics with extended culture. To determine hormonal responsiveness of cultured melanocytes and investigate regulation of melanocortin 1 receptor expression, cultured melanocytes were treated with increasing concentrations of ${\alpha}$-melanocyte stimulating hormone. Treatment with ${\alpha}$-melanocyte stimulating hormone increased melanocortin receptor 1 mRNA in a dose dependent fashion. The results demonstrated culture of pure populations of alpaca melanocytes to 10 passages and illustrate the potential utility of such cells for studies of intrinsic and extrinsic regulation of genes controlling pigmentation and coat color in fiber-producing species.

Analysis of Immunomodulating Gene Expression by cDNA Microarray in $\beta$-Glucan-treated Murine Macrophage

  • Sung, Su-Kyong;Kim, Ha-Won
    • 한국응용약물학회:학술대회논문집
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    • 한국응용약물학회 2003년도 Annual Meeting of KSAP : International Symposium on Pharmaceutical and Biomedical Sciences on Obesity
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    • pp.98-98
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    • 2003
  • ${\beta}$-(1,3)-D-Glucans have been known to exhibit antitumor and antimicrobial activities. The presence of dectin-1,${\alpha}$, ${\beta}$-glucan receptor of dendritic cell, on macrophage has been controvertial. RT-PCR analysis led to the detection of dectin-1${\alpha}$ and ${\beta}$ in murine macrophage Raw264.7 cell line. Among the various organs of mouse, dectin-1${\alpha}$ and ${\beta}$ were detected in the thymus, lung, spleen, stomach and intestine. To analyze gene expression modulated by ${\beta}$-glucan treated murine Raw264.7 macrophage, total mRNA was applied to cDNA microarray to interrogate the expression of 7,000 known genes. cDNA chip analysis showed that ${\beta}$-glucan of P. osteatus increased gene expressions of immunomodulating genes, membrane antigenic proteins, chemokine ligands, complements, cytokines, various kinases, lectin associated genes and oncogenes in Raw 264.7 cell line. When treated with ${\beta}$-glucan of P. osteatus and LPS, induction of gene expression of TNF-${\alpha}$ and IFN-R1 was confirmed by RT-PCR analysis. Induction of TNF-R type II expression was confirmed by FACS analysis. IL-6 expression was abolished by EDTA in ${\beta}$-glucan and LPS treated Raw264.7 cell line, indicating that ${\beta}$-glucan binds to dectin-l in a Ca$\^$++/ -dependent manner. To increase antitumor efficacy of ${\beta}$-glucan, ginsenoside Rh2 (GRh2) was co-treated with ${\beta}$-glucan in vivo and in vitro tests. IC$\sub$50/ values of GRh2 were 20 and 25 $\mu\textrm{g}$/$m\ell$ in SNU-1 and B16 melanoma F10 cell line, respectively. Co-treatment with ${\beta}$-glucan and GRh2 showed synergistic antitumor activity with cisplatin and mitomycin C both in vitro and in vivo. Single or co-treatment with ${\beta}$-glucan and GRh2 increased tumor bearing mouse life span. Co-treatment with ${\beta}$-glucan and GRh2 showed more increased life span with mitomycin C than that with cisplatin. Antitumor activities were 67% and 72 % by co-injection with ${\beta}$-glucan and GRh2 in the absence or presence of mitomycin C, respectively.

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Effects of Iron, Chelators and Nitrate Concentration on in vivo Fluorescence and Nitrate Reductase of the Red Tide Organism Amphidinium carterae

  • Yang, Sung-Ryull;Song, Hwan-Seok;Pae, Se-Jin;Huh, Sung-Hoi
    • Journal of the korean society of oceanography
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    • 제34권1호
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    • pp.49-57
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    • 1999
  • A red tide organism, Amphidinium carterae was incubated under different iron/chelator and nitrate concentrations to investigate the factors controlling the growth. The chelation capacity played a critical role in regulating the nitrate reductase (NR) activity and in vivo fluorescence of this organism. However, there was a significant difference between the NR activity and in vivo fluorescence in response to trace metals and chelator treatments. In vivo fluorescence was the highest in FeEDTA 10 ${\mu}$M treatments and the lowest in DTPA 10 ${\mu}$M treatments. This indicates that the availability of the trace metal is important in regulating the in vivo fluorescence of this photosynthetic microalgae In contrast, NR activity showed the highest values in trace metal enriched treatments, and trace metal + DTPA treatments showed fairly high NR activities. This suggests that DTPA treatment did not hinder the NR activity as much as it did in vivo fluorescence. In vivo fluorescence and NR activity increased with nitrate concentration of up to 50 ${\mu}$M and remained relatively constant or the rate of increase decreased above that concentration, indicating that initial nitrate concentration of higher than a certain level would not accelerate the growth of A. carterae. Further investigation is needed to elucidate the reason for the difference in timing sequence between the NR and in vivo fluorescence in response to different metal treatments and chelation capacity.

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Biological effects of dietary probiotics on blood characteristics in Hanwoo heifers subjected to lipopolysaccharide (LPS) challenge

  • Park, Bo Hye;Kim, Ui Hyung;Jang, Sun Sik;Yang, Seung Hak;Lee, Eun Mi;Kang, Dong Hun;Kwon, Eung Gi;Chung, Ki Yong
    • 농업과학연구
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    • 제43권5호
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    • pp.818-827
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    • 2016
  • This study hypothesized that dietary feed additive containing probiotics alter either immune-related serum substances or serum metabolites in Hanwoo heifers. A probiotic treatment was given at 0.5% top-dressing of concentrate diet for 6 months. The change of immunological indicators in the blood was analyzed under LPS (Lipopolysaccharide) challenge. One day before administration of LPS, all heifers were fitted with an indwelling jugular vein catheter for serial blood collections. Both a serum tube and an EDTA-coated tube were collected at 30-min intervals from - 2 to 8 hours relative to the LPS challenge at time 0 ($1{\mu}g/kg$ of BW). Serum was used for analyzing albumin (ALB), glucose (GLU), total protein (TP), triglycerides (TG), phosphorus (IP), and non-esterified fatty acids (NEFA). Plasma was used for analyzing white blood cell (WBC), red blood cell (RBC), platelet (PLT) and inflammation-related factors (NE, LY, MO, EO, BA, Hb, HCT, MCV, MCH, MCHC, RDW, MPV). There were significant differences in ALB, GLU, TG, IP, and NEFA concentration with the passage of hours post challenge (p < 0.05). The level of ALB, GLU, TG, and IP showed significant difference (p < 0.05) between treatments. However, none of the data showed interaction between time and treatments (p > 0.05). The level of WBC, EO, LY, and MO were reduced after LPS challenge (p > 0.05). In conclusion, LPS challenge after dietary supplementation of probiotics changed the levels of both serum metabolites and inflammation-related factors. The increase of GLU and TG indicated a probiotics-positive response under LPS challenge (p < 0.05).

Efficient removal of radioactive waste from solution by two-dimensional activated carbon/Nano hydroxyapatite composites

  • El Said, Nessem;Kassem, Amany T.
    • Membrane and Water Treatment
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    • 제9권5호
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    • pp.327-334
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    • 2018
  • The nano/micro composites with highly porous surface area have attracted of great interest, particularly the synthesis of porous and thin film sheets of high performance. In this paper, an easy method of cost-effective synthesis of thin film ceramic fiber membranes based on Hydroxyapatite, and activated carbon by turned into studied to be applied within the service-facilitated the transport of radioactive waste such as $^{90}Sr$, $^{137}Cs$ and $^{60}Co$) as activated product of radioisotopes from ETRR-2 research reactor and dissolved in 3M $HNO_3$, across a thin flat-sheet supported liquid membrane (TFSSLM). Radionuclides are transported from alkaline pH values. The presence of sodium salts in the aqueous media improves in $HNO_3$, the lowering of permeability because the initial $HNO_3$ concentration is improved. The study some parameters on the thin sheet ceramic supported liquid membrane. EDTA as stripping phase concentration, time of extraction and temperature were studied. The study of maximum permeability of radioisotopes for all parameters. The pertraction of a radioactive waste solution from nitrate medium were examined at the optimized conditions. Under the optimum experimental 98.6-99.9% of $^{90}Sr$, 79.65-80.3% of $^{137}Cs$ and $^{60}Co$ 45.5-55.5% in 90-110 min with were extracted in 10-30 min, respectively. The process of diffusion in liquid membranes is governed by the chemical diffusion process.

체내 수정된 Mouse 1-세포배의 체외발생에 미치는 혈관내피세포주(tHUE-2세포) 배양액의 영향 (Effect of Conditioned Medium of Human Endothelial Cell Line(tHUE-2 cell) on In Vitro Development of Mouse 1-cell Embryos In Vitro Fertilized)

  • 박흠대;이훈택;정길생
    • 한국가축번식학회지
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    • 제19권2호
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    • pp.81-88
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    • 1995
  • Culture medium (ASF-301) of tHUE-2 cell, human endothelial cell line, and culture medium of these cells (conditioned medium : CM) which affect embryonic development of in vivo fertilized 1-cell embryos of mouse were examined. Two-cell stage block of mouse embryos was overicomed in ASF-301 and CM without EDTA, which usually added in basic medium (modified Whitten Medium: MWM, control) to overcome the 2-cell stage block. The developmental rates of embryos to the blastocyst stage were significantly increased in MWM containing 12.5% of growth factors added to ASF-301 (10mg/ $\ell$ transferrin, 1mg/$\ell$ insulin, 0.01mg/$\ell$ EGF) than those of 100% addition and control, 78.0% vs 20.8 and 52.3%, respectively (P<0.05), but the growth factors was not affected the hatching rate of blastocyst. Using ASF-301 or CM which was not treated, embryonic development into the blastocyst and hatched blastocyst stages were not affected. However, proportions of embryonic development into the blastocyst and hatched blastocyst stages were significantly higher in dilution (ASF-301 1:10; CM 1:3~1:6) than those in control (P,0.05). In ASF-301 dialyzed M.W.<10000 dialysis membrane, the developmental rate upto the hatched blastocyst stage was significantly increased, compared to ASF-301 which was not dialyzed (P<0.05), and hatching rate of blastocyst of these group was singnificantly increased than those in MWM (P<0.05). Compared to CM which was not dialyzed, however, in dialyzed CM was significantly decreased, compared to untreated CM (P<0.05), especially any hatched blastocyst was not appeared. As a result of these experiments indicated that a kind or porper treatment such as a dilution of complex synthetic cell culture medium and conditioned medium, and that a optimal concentration of growth factors are usuful for embryo cultrue in vitro.

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미노싸이클린 연고의 근육 내 투여 후 조직 및 혈중 약물농도의 분석 (High-Performance Liquid Chromatography를 이용하여) (ANALYSIS OF MINOCYCLINE IN TISSUE AND SERUM AFTER LOCAL APPLICATION OF MINOCYCLINE OINTMENT (by High-Performance Liquid Chromatography))

  • 황혜욱;이상철;김성진
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • 제25권4호
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    • pp.304-310
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    • 1999
  • This study was designed to find the effect of Minocycline loaded microcapsule applied locally to tissue by measuring drug concentration in tissue and serum by HPLC and to achieve optimal drug delivery system and duration to a specific target site. Control group were administrated minocycline intramuscularly twice a day with $0.2{\mu}g/100g$ for 1 to 10 days. In experimental group, surgical wound was created on Rt. cheek and then minocycline loaded microcapsule was applied into the space between superficial and deep layer of masseter muscle. Animals were sacrificed at 1, 3, 5, 7, 10 days after initial administration, blood was obtained from heart and right masseter muscle was excised. Blood sample was centrifuged at 3000rpm for 15min. Tissue sample was homogenized, left at room temperature for 48hr and centrifuged at 4000g for 5min. Supernatant was completely dried and dissolved in distilled water. Analysis was conducted using a ${\mu}Bondapack$ C18 column. The mobile phase was 0.2M Ammonium Oxalate/0.1M EDTA/DMF=11/4/5 solution, which was injected into the column and detected with photodiode detector at 344nm wavelength. The results were as follows : 1. This method was reliable, could be replicated and suitable for minocycline analysis in tissue as well as serum. 2. In tissue, concentration of minocycline of experimental group was higher than that of control group for 5days. 3. Except 1 day, concentration of minocycline in serum of experimental group was lower than that of control group. 4. Concentration of minocycline in tissue was much higher than that in serum. From these results, minocycline loaded microcapsule might be effective tool for local drug delivery system might be useful for treatment of infections of oral and maxillofacial region and management of infected surgical wound, minimizing systemic effects.

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CO2 laser조사가 성견치수에 미치는 영향에 관한 실험적 연구 (THE EFFECT OF CO2 LASER ON DENTAL PULP OF DOG)

  • 김희중;이찬영;이승종;이정석
    • Restorative Dentistry and Endodontics
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    • 제13권1호
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    • pp.7-19
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    • 1988
  • The object of this paper was to investigate the histopatological changes on dog's pulp under cavitation by irradiation of the $CO_2$ laser. The subjects were derived from four dogs, and irradiated 113.23 J/$mm^2$, 283.09 J/$mm^2$, 566.08 J/$mm^2$ in Group I, II, and III respectively. The dogs were sacrificed immediately, 24 hour, 72 hour and 1 week after $CO_2$ laser treatment. For light microscopic examination, routine H-E and PAS stains were employed. For electron microscopic observation, the teeth were fixed in 1% paraformaldehyde and 1% glutaraldehyde, decalcified teeth in 10% EDTA were stained by uranyl acetate and lead citrate. The observation was made with a Hitachi H-500 model electron microscope. The following results were obtained in this study: 1. At the early stage of the experimental sub-groups-immediately, 24 hour, 72 hour samples of Group I, II and III-coagulation necrosis and hyperemia were observed in odontoblastic and subodontoblastic pulpal layer. 2. At the 1 week sub-group of Group I, II, regenerative hyperplasia of the odontoblasts without coagulation necrosis were revealed, in addition to thickened predentin. On he other hand coagulation necrosis and atrophic change accompanying with hyperplasia were found at the 1 week sub-group of Group III. 3. Ultrastructurally, the odontoblasts appeared nuclear degeneration, vacuolar change of cytoplasmic organelles and rupture of plasma membrane at the early stage of the experimental period of all groups. 4. Under spectrohelioscopic examination, regenerative odontobalsts were seen at the 1 week specimens of Group I, II and III. 5. The pulpal response occured at 113-566 J/$mm^2$. The pathologic change of pulp tissue occured at the early experimental period but regeneration of odontoblasts could be seen after 1 week.

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Chemical Modification of 5-Lipoxygenase from the Korean Red Potato

  • Kim, Kyoung-Ja
    • BMB Reports
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    • 제33권2호
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    • pp.172-178
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    • 2000
  • The lipoxygenase was purified 35 fold to homogeneity from the Korean red potato by an ammonium sulfate precipitation and DEAE-cellulose column chromatography. The simple purification method is useful for the preparation of pure lipoxygenase. The molecular weight of the enzyme was estimated to be 38,000 by SDS-polyacrylamide gel electrophoreses and Sepharose 6B column chromatography. The purified enzyme with 2 M $(NH_4)_2SO_4$ in a potassium phosphate buffer, pH 7.0, was very stable for 5 months at $-20^{\circ}C$. Because the purified lipoxygenase is very stable, it could be useful for the screening of a lipoxygenase inhibitor. The optimal pH and temperature for lipoxygenase purified from the red potato were found to be pH 9.0. and $30^{\circ}C$, respectively. The Km and Vmax values for linoleic acid of the lipoxygenase purified from the red potato were $48\;{\mu}M$ and $0.03\;{\mu}M$ per minute per milligram of protein, respectively. The enzyme was insensitive to the metal chelating agents tested (2 mM KCN, 1 and 10mM EDTA, and 1 mM $NaN_3$), but was inhibited by several divalent cations, such as $Cu^{++}$, $Co^{++}$ and $Ni^{++}$. The essential amino acids that were involved in the catalytic mechanism of the 5-lipoxygenase from the Korean red potato were determined by chemical modification studies. The catalytic activity of lipoxygenase from the red potato was seriously reduced after treatment with a diethylpyrocarbonate (DEPC) modifying histidine residue and Woodward's reagent (WRK) modifying aspartic/glutamic acid. The inactivation reaction of DEPC (WRK) processed in the form of pseudo-first-order kinetics. The double-logarithmic plot of the observed pseudo-first-order rate constant against the modifier concentration yielded a reaction order 2, indicating that two histidine residues (carboxylic acids) were essential for the lipoxygenase activity from the red potato. The linoleic acid protected the enzyme against inactivation by DEPC(WRK), revealing that histidine and carboxylic amino acids residues were present at the substrate binding site of the enzyme molecules.

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