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PERIPHERAL NERVE REGENERATION USING POLYGLYCOLIC ACID CONDUIT AND BRAIN-DERIVED NEUROTROPHIC FACTOR GENE TRANSFECTED SCHWANN CELLS IN RAT SCIATIC NERVE (BDNF 유전자 이입 슈반세포와 PGA 도관을 이용한 백서 좌골신경 재생에 관한 연구)

  • Choi, Won-Jae;Ahn, Kang-Min;Gao, En-Feng;Shin, Young-Min;Kim, Yoon-Tae;Hwang, Soon-Jeong;Kim, Nam-Yeol;Kim, Myung-Jin;Jo, Seung-Woo;Kim, Byung-Soo;Kim, Yun-Hee;Kim, Soung-Min;Lee, Jong-Ho
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.30 no.6
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    • pp.465-473
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    • 2004
  • Purpose : The essential triad for nerve regeneration is nerve conduit, supporting cell and neurotrophic factor. In order to improve the peripheral nerve regeneration, we used polyglycolic acid(PGA) tube and brain-derived neurotrophic factor(BDNF) gene transfected Schwann cells in sciatic nerve defects of SD rat. Materials and methods : Nerve conduits were made with PGA sheet and outer surface was coated with poly(lactic-co-glycolic acid) for mechanical strength and control the resorption rate. The diameter of conduit was 1.8mm and the length was 17mm Schwann cells were harvested from dorsal root ganglion(DRG) of SD rat aged 1 day. Schwann cells were cultured on the PGA sheet to test the biocompatibility adhesion of Schwann cell. Human BDNF gene was obtained from cDNA library and amplified using PCR. BDNF gene was inserted into E1 deleted region of adenovirus shuttle vector, pAACCMVpARS. BDNF-adenovirus was multiplied in 293 cells and purified. The BDNF-Adenovirus was then infected to the cultured Schwann cells. Left sciatic nerve of SD rat (250g weighing) was exposed and 14mm defects were made. After bridging the defect with PGA conduit, culture medium(MEM), Schwann cells or BDNF-Adenovirus infected Schwann cells were injected into the lumen of conduit, respectively. 12 weeks after operation, gait analysis for sciatic function index, electrophysiology and histomorphometry was performed. Results : Cultured Schwann cells were well adhered to PGA sheet. Sciatic index of BDNF transfected group was $-53.66{\pm}13.43$ which was the best among three groups. The threshold of compound action potential was between 800 to $1000{\mu}A$ in experimental groups which is about 10 times higher than normal sciatic nerve. Conduction velocity and peak voltage of action potential of BDNF group was the highest among experimental groups. The myelin thickness and axonal density of BDNF group was significantly greater than the other groups. Conclusion : BDNF gene transfected Schwann cells could regenerate the sciatic nerve gap(14mm) of rat successfully.

A Simultaneously Analytical Method of Phthalate and Adipate Plasticizers in Food Packaging by Dual-Column GC-FID System (Dual-Column GC-FID System을 이용한 식품 포장재 중 Phthalate류 및 Adipate류 가소제의 동시 분석법)

  • Kang Gil-Jin;Kwak In-Shin;Eom Mi-Ok;Jeon Dea-Hoon;Kim Hyung-il;Sung Jun-Hyun;Choi Jung-Mi;Kim Eun-Kyung;Lee Young Ja
    • Journal of Food Hygiene and Safety
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    • v.20 no.4
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    • pp.277-283
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    • 2005
  • A plasticizer is a substance which is added to a material to improve its processability, flexibility and stratchability. Phthalates and adipates are the most frequently used plasticizers of poly(vinyl chloride) (PVC). However, they can migtate into food from PVC food packaging, and some of them are especially suspected as endocrine disruptors. In this study, Simultaneous analysis of 13 phthalates and 9 adipates were carried out by dual-column gas chromatography system equipped wi two FID detectors for rapid confirmation and quantification. The Proposed method was validated with > 0.993 of linearity in the ranges of 10-500 mg/l, < $3.5\%$ RSD of reproducability in 10 inter-days sample preparations, and > $98.1\%$ of recoveries for all the plasticizers. DEHA was detected in all the 3 PVC wraps at levels of 176.9-198.5mg/g. Among the 51 samples of PVC gaskets, the targeted plasticizers were detected in 41 samples. Of these plasticizer detected samples,40 contained DIDP at the levels of 157.3-374.7 mg/g and one contained DMP at the levels of 165.6 mg/g. Also, some plasticizers were detected in other packaging materials such as PET, PP, PE, Pulp. But it might be attributed to contamination in manufacturing.

Analysis of the Seasonal Change of the Proximate Composition and Taste Components in the Conger Eels (Conger myriaster) (붕장어(Conger myriaster)의 일반성분 및 맛 성분의 계절별 변화 분석)

  • Ryu, Keun-Young;Shim, Sung-Lye;Kim, Won;Jung, Min-Seok;Hwang, In-Min;Kim, Jun-Hyeong;Hong, Cheul-Hee;Jung, Chan-Hee;Kim, Kyong-Su
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.38 no.8
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    • pp.1069-1075
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    • 2009
  • Conger eel (Conger myriaster) is used as a well-being food in the foodservice industry in Korea. We analyzed not only the proximate composition but also the taste components affecting conger eel, which are fatty acids, nucleotides, amino acids, etc. Concerning the composition, the crude lipid was the lowest in summer at 3.2%, which is considered due to spawning period. The major fatty acids were $C_{16:0},\;C_{16:1},\;C_{18:0},\;C_{18:1},\;C_{20:5},\;and\;C_{22:6}$. The $C_{18:1}$ content was the highest among the fatty acids and the content varied between 36.76 and 45.11% by season. Seasonal change in the content of poly-unsaturated fatty acids was increased from spring to winter in conger eel. Among the nucleotides, the contents of IMP (3.617$\sim$5.524 $\mu$mol/g) and Hx (0.913$\sim$2.238 $\mu$mol/g), which is closely related to taste, and the concentrations of IMP and Hx were the highest (7.219 $\mu$mol/g) in winter, and HxR (0.625$\sim$1.652 $\mu$mol/g) was higher than ATP (0.058$\sim$0.083 $\mu$mol/g), ADP (0.145$\sim$0.161 $\mu$mol/g), and AMP (0.166$\sim$0.179 $\mu$mol/g). In conger eels, the major total and free amino acids were glutamic acid (14,178.7$\sim$7,802.6 mg%), aspartic acid (4,669.2$\sim$8,259.0 mg%), lysine (4,198.3$\sim$7,540.8 mg%), leucine (3,843.6$\sim$6,782.1 mg%), and histidine (199.6$\sim$644.4 mg%), glycine (94.8$\sim$152.2 mg%), alanine (35.3$\sim$71.2 mg%), glutamic acid (44.1$\sim$70.6 mg%), respectively, but the concentration of amino acids was different by season. The content of free amino acids, which is related to the taste component, was detected as high in summer and winter at 1179.2 and 1,605.2 mg%, respectively.

Relationship between Steady Flow and Dynamic Rheological Properties for Viscoelastic Polymer Solutions - Examination of the Cox-Merz Rule Using a Nonlinear Strain Measure - (점탄성 고분자 용액의 정상유동특성과 동적 유변학적 성질의 상관관계 -비선헝 스트레인 척도를 사용한 Cox-Merz 법칙의 검증-)

  • 송기원;김대성;장갑식
    • The Korean Journal of Rheology
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    • v.10 no.4
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    • pp.234-246
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    • 1998
  • The objective of this study is to investigate the correlation between steady shear flow (nonlinear behavior) and dynamic viscoelastic (linear behavior) properties for concentrated polymer solutions. Using both an Advanced Rheometic Expansion System(ARES) and a Rheometics Fluids Spectrometer (RFS II), the steady shear flow viscosity and the dynamic viscoelastic properties of concentrated poly(ethylene oxide)(PEO), polyisobutylene(PIB), and polyacrylamide(PAAm) solutions have been measured over a wide range of shear rates and angular frequencies. The validity of some previously proposed relationships was compared with experimentally measured data. In addition, the effect of solution concentration on the applicability of the Cox-Merz rule was examined by comparing the steady flow viscosity and the magnitude of the complex viscosity Finally, the applicability of the Cox-Merz rule was theoretically discussed by introducing a nonlinear strain measure. Main results obtained from this study can be summarized as follows : (1) Among the previously proposed relationships dealt with in this study, the Cox-Merz rule implying the equivalence between the steady flow viscosity and the magnitude of the complex viscosity has the best validity. (2) For polymer solutions with relatively lower concentration, the steady flow viscosity is higher than the complex viscosity. However, such a relation between the two viscosities is reversed for highly concentrated polymer solutions. (3) A nonlinear strain measure is decreased with increasing stran magnitude, after reaching the maximum value in small strain range. This behavior is different from the theoretical prediction demonstrating the shape of a damped oscillatory function. (4) The applicability of the Cox-Merz rule is influenced by the $\beta$ value, which indicates the slope of a nonlinear stain measure (namely, the degree of nonlinearity) at large shear deformations. The Cox-Merz rule shows better applicability as the $\beta$ value becomes smaller.

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Disturbance of $\alpha$-Amylase Secretion from Bacillus amyloliquefaciens Cells by the Treatment of Puromycin and Magnesium (Bacillus amyloliquefaciens에서 Puromycin 과 Magnesium에 의한 $\alpha$-Amylase 의 분비저해)

  • 안순자;김순옥;이동희;송방호
    • Microbiology and Biotechnology Letters
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    • v.17 no.5
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    • pp.412-420
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    • 1989
  • To know how the ribosomes involved in secretory protein synthesis were attached to the cytoplasmic membrane in Bacillus amyloliquefaciens, the cells were treated with puromycin combinated with magnesium at the logarithmic phase, and the variation of cell-bound and extracellular $\alpha$-amylase activity was assayed for determining the $\alpha$-amylase translocation blocking through the cytoplasmic membrane. In the abnormal $\alpha$-amylase producing mutant in which the C-terminal of the $\alpha$-amylase structure was deleted, B. umytotiquefaciens CH10-2, the $\alpha$-amylase was translocated normally through the cytoplasmic membranes, and the translocation blocking by puromycin was revealed to have a similar pattern as that in the wild type. This means that the C-terminal part of the enzyme structure may not have a signal for secretion. The cell death of the logarithmic phase cells in both strains was not affected much under 20$\mu\textrm{g}$/$m\ell$ of puromycin, however, the $\alpha$-amylase translocation was blocked markedly under less than 10$\mu\textrm{g}$/$m\ell$ of the puromycin concentration. The blocking of the enzyme secretion by puromycin may be due to the detachment of the ribosomes from cytoplasmic membranes by disturbing the nascent polypeptide synthesis. Further evidence for confirming this was that the detachment was increased in 50 mM of magnesium ion because the extracellular $\alpha$-amylase activity was decreased more under this condition. If the cells were treated with trypsin combinated with Iysozyme, the extracellular $\alpha$-amylase activity from the cultured medium was reduced markedly, however, the activity from the cells treated with trypsin only was not reduced. This means that the nascent polypeptides protruding from the cytoplasmic membrane were sensitive to the trypsin digestion, whereas the matured ones were not. Therefore, the protruding polypeptides from the cytoplasmic membranes may be truncated by trypsin before forming their final tertiary structures by folding in the cell wall layer.

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Survey of Physicochemical Methods and Economic Analysis of Domestic Wastewater Treatment Plant for Advanced Treatment of Phosphorus Removal (총인 수질기준강화를 위한 국내 하수종말처리장의 물리화학적처리 특성조사 및 경제성 분석)

  • Park, Hye-Young;Park, Sang-Min;Lee, Ki-Cheol;Kwon, Oh-Sang;Yu, Soon-Ju;Kim, Shin-Jo
    • Journal of Korean Society of Environmental Engineers
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    • v.33 no.3
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    • pp.212-221
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    • 2011
  • Wastewater treatment plants (WWTPs) are required to meet the reinforced discharge standards which are differentiated as 0.2, 0.3 and 0.5 mg-TP/L for the district I, II and III, respectively. Although most of WWTPs are operating advanced biological phosphorus removal system, the supplementary phosphorus treatment facility using chemical addition should be required almost at all WWTPs. Therefore, water quality data from several exemplary full-scale plants operating phosphorus treatment process were analyzed to evaluate the reliability of removal performance. Additionally, a series of jar tests were conducted to find optimal coagulants dose for phosphorus removal by chemical precipitation and to describe characteristics of the reaction and sludge production. Chemical costs and the increasing sludge volume in physicochemical phosphorus removal process were estimated based on the results of jar tests. The minimum coagulant (aluminium sulfate and poly aluminium chloride) doses to keep TP concentration below 0.5 and 0.2 mg/L were around 25 and 30 mg/L (as $Al_2O_3$), respectively, in the mixed liquor of activated sludge. In the tertiary treatment facility, relatively lower coagulant doses of 1/12~1/3 the minimum doses for activated sludge were required to achieve the same TP concentrations of 0.2~0.5 mg/L. Increase in suspended solids concentration due to chemical precipitates in mixed liquor was estimated at 10~11%, compared to the concentration without chemical addition. When coagulant was added into mixed liquor, chemical (aluminium sulfate) cost was estimated to be 4~10 times higher than in secondary effluent coagulation/separation process. Sludge production to be wasted was also 4~10 times higher than secondary effluent coagulation/separation process.

Effect of Cryoprotectants on Quality Properties of Chicken Breast Surimi Manufactured by pH Adjustment (냉동변성방지제 조건에 따른 닭가슴살 수리미의 저장특성)

  • Jin, S.K.;Kim, I.S.;Choi, Y.J.;Park, G.B.;Yang, H.S.;Kim, B.G.
    • Journal of Animal Science and Technology
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    • v.50 no.1
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    • pp.99-110
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    • 2008
  • This study was conducted to determine the effect of pH adjustment and the addition of cryoprotectants on the storage properties of chicken breast surimi. Surimi as a control was prepared with Alaska pollack by two times of washing treatment and addition of cryoprotectants(4% sugar, 5% sorbitol and 0.3% polyphosphate). Three types of surimi were manufactured by different cryoprotectant conditions(T1: 5% sorbitol+0.3% polyphosphate, T2: 4% sugar+5% sorbitol+0.3% polyphosphate, T3: 2% salt+4% sugar+5% sorbitol+0.3% polyphosphate) and frozen after adjusting the surimi to pH 11.0. The amino acid and saturated fatty acid composition were significantly higher in the control. EAA(essential amino acid), FAA(amino acid in relation to flavor), SAAA(amino acid in relation to saccarinity) and FRAA (fragrant amino acid) were significantly higher in the control. The TBARS(thiobarbituric acid reactive substances) and VBN(volatile basic nitrogen) were increased with storage times. TBARS and VBN values were significantly higher in the control and T3 than the other treatments. The VBN was significantly higher in control and T3 than other surimi samples. Chicken breast surimi adjusted to pH 11.0 had higher in microorganisms than the control, and T1 sample had the highest total plate count. Sensory evaluations were significantly higher in the control and T3 than the other samples. Especially, the overall acceptability of T3 was similar to the control.

Ultrastructural analysis and quantification of autophagic vacuoles in wild-type and atg5 knockout mouse embryonic fibroblast cells (정상 및 atg5 유전자 제거 섬유아세포에서 자가포식체의 미세구조 및 이들의 정량적 분석)

  • Choi, Suin;Jeon, Pureum;Huh, Yang Hoon;Lee, Jin-A
    • Analytical Science and Technology
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    • v.31 no.5
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    • pp.208-218
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    • 2018
  • Autophagy is a cellular process whereby cytosolic materials or organelles are taken up in a double-membrane vesicle structure known as an autophagosome and transported into a lysosome for degradation. Although autophagy has been studied at the genetic, cellular, or biochemical level, systematic ultrastructural quantitative analysis of autophagosomes during the autophagy process by using transmission electron microscopy (TEM) has not yet been reported. In this study, we performed ultrastructural analysis of autophagosomes in wild-type (WT) mouse embryonic fibroblasts (MEFs) and autophagy essential gene (atg5) knockout (KO) MEFs. First, we performed ultrastructural analysis of autophagosomes in WT MEFs compared to atg5 KO MEFs in basal autophagy or starvation-induced autophagy. Although we observed phagopore, early, late autophagosomes, or autolysosomes in WT MEFs, atg5 KO MEFs had immature autophagosomes that showed incomplete closure. Upon starvation, late autophagosomes accumulated in WT MEFs while the number of immature autophagosomes significantly increased in atg5 KO MEF indicating that atg5 plays an important role in the maturation of autophagosomes. Next, we examined autophagosomes in the cell model expressing polyQ-expanded N-terminal fragment of huntingtin. Our TEM analysis indicates that the number of late autophagosomes was significantly increased in the cells expressing the mutant huntingtin, indicating that improving the fusion of autophagosome with lysosome may be effective to enhance autophagy for the treatment of Huntington's disease. Taken together, the results of our study indicate that ultrastructural and quantitative analysis of autophagosomes using TEM can be applied to various human cellular disease models, and that they will provide an important insight for cellular pathogenesis of human diseases associated with autophagy.

Induction of G2/M Arrest and Apoptosis by the Methanol Extract of Typha orientalis in Human Colon Adenocarcinoma HT29 Cells (포황 메탄올 추출물에 의한 인체 대장암 세포주 HT29의 G2/M Arrest 및 Apoptosis 유발)

  • Jin, Soojung;Yun, Seung-Geun;Oh, You Na;Lee, Ji-Young;Park, Hyun-Jin;Jin, Kyong-Suk;Kwon, Hyun Ju;Kim, Byung Woo
    • Microbiology and Biotechnology Letters
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    • v.41 no.4
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    • pp.425-432
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    • 2013
  • Typha orientalis, also known as bulrush or cattail, is a perennial herbaceous plant found in freshwater wetlands and has been widely used in constructed wetlands for wastewater treatment. Recent data has revealed that SH21B, a mixture composed of seven herbs including T. orientalis, exhibited an anti-adipogenic activity by the inhibition of the expression of adipogenic regulators. However, the anti-cancer effect of T. orientalis and its molecular mechanisms remain unclear. In this study, we evaluated the anti-cancer effect and its mechanism in the methanol extract of T. orientalis (METO) on human colon carcinoma HT29 cells. It was found that METO treatment showed cytotoxic activity in a dose-dependent manner, and induced G2/M cell cycle arrest and apoptosis in HT29 cells. The induction of G2/M arrest by METO was associated with the up-regulation of phospho-Cdc2 (Tyr15), an inactive form of Cdc2 and the down-regulation of Cdc25c phosphatase. METO also induced tumor suppressor p53 and cyclin-dependent kinase inhibitor p21 (WAF1/CIP1) expression. In addition, METO-induced apoptosis was characterized by the proteolytic activation of caspase-3, degradation of poly ADP ribose polymerase (PARP), and up-regulation of death receptor FAS and pro-apoptotic Bax expression. Collectively, these results indicate that the cell cycle inhibition and apoptosis induction of METO in HT29 cells allows for the possibility of its use in anti-cancer therapies.

The Effect of Water Immersion on the Surface Strength and the Flexural Strength of the Acrylic Resin for Occlusal Appliances (교합장치용 아크릴레진의 표면경도와 굴곡강도에 대한 침수의 영향)

  • Lee, Hoy-Youn;Im, Yeong-Gwan;Kim, Byung-Gook;Lim, Hoi-Soon;Kim, Jae-Hyung
    • Journal of Oral Medicine and Pain
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    • v.35 no.1
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    • pp.75-81
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    • 2010
  • By repeating nocturnal bruxism occlusal appliance's wearing condition that is used to cure temporomandibular disorders into the vitro experiment, research aims to find out how moisture infiltrated and drying cycle process affects on the surface microhardness of the resin for occlusal appliance and flexural strength. By utilizing resin for occlusal appliance which is the main component of poly methyl methacrylate, bar shaped sample was produced. For the resin sample utilized as the controlled group 1, the sample was infiltrated in the moisture for 7 days in the temperature of 37C. Then, the resin sample of the controlled group 2 was maintained in a dry condition for 7 days in the normal temperature. After that, each sample's surface microhardness and flexural strength were measured. For the resin sample that is utilized as the experimental sample, the sample was infiltrated in the moisture for 7 days in the temperature of 37C. Then, it was inundated for 8 hours a day and dried in the normal temperature for 16 hours with the continuous process of moisture infiltration and dry cycle process for 30 days. During this cycle process, sample's surface and flexural strength were measured in the 1st day, 7th day, and 30th day. Then, it was statistically analyzed to find out the difference of controlled and experiment group's surface microhardness and flexural strength. Results are 1. For the experimental and controlled group's surface microhardness of the resin for the occlusal appliance, it did not show any significant differences after moisture infiltration and dry cycle process repetition. 2. In case of the flexural strength for resin for the occlusal appliance, experimental group with moisture infiltration and dry cycle for 30 day process had greater effect than the experimental group at the 1st day and controlled group These results can be considered to be utilized from the patients of the temporomandibular disorders towards occlusal appliance used and maintained method.