Park, Dae-Woo;Kang, Hyun-Mi;Nam, Byeong-Jik;Jang, Sung-Yoon;Ham, Chul-Hee
보존과학연구
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s.31
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pp.43-57
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2010
This study intends to provide quantitative data about the extraction characteristics of major elements of earthenware by executing soaking test of cleaning agents. It aims at providing basic data for the stability assessment when applying the cleaning agents for conserving earthenware. The data will be extracted from the analysis of co-relationship between the physical characteristics and the ion extraction characteristics. XRD analysis displayed that AT-1, AT-2 and AT-3 which did not generate mullite were fired at lower than 1,000 whereas AT-3 and AT-5 that included mullite were higher than 1,000. The degree of absorption was AT-4 > AT-2 > AT-1 > AT-3 > AT-5 in order and the correlation between the degree of absorption and firing temperature of earthenware displayed a positive correlation. Extraction amount of oxalic acid which was used for the removing iron oxide was AT-1 > AT-2 AT-4 > AT-3 > AT-5 in order. and the ion extraction data displayed that there is a positive correlation with absorption level. However AT-1 and AT-2 which were fired at lower temperature showed that there was no correlation between the ion extraction characteristics and absorption level. Ion extraction of citric acid produced little amount compared with the one of oxalic acid, yet it caused less damage to earthenware than oxalic acid when it applied. The result of ion extraction level in the absorption test displayed that Fe had higher level than in Si, Al from the test for both oxalic acid and citric acid. Based on the regression analysis of the data from the previous studies, the physical characteristics of the earthenware and ion extraction level, further studies will be conducted on the predicting technique on the extraction characteristics of major elements of earthenware samples for the conservation in future.
A Pseudomonas sp. strain that is capable of utilizing dicarboxylic acids as a sole carbon source was isolated from activated sludge by using the enrichment culture technique. This organism accumulated polyhydroxyalkanoates (PHAs) with an unusual pattern of monomer units that depends on the carbon sources used. Polyhydroxybutyrate (PHB) homopolyester was synthesized from glucose or small $C_{-even}$ alkanoic acids, such as butyric acid and hexanoic acid. Accumulation of PHB homopolyester was also observed in the cells grown on $C_{-odd}$ dicarboxylic acids, such as heptanedioic acid and nonanedioic acid as the sole carbon sources. In contrast, a copolyester consisting of 6 mol% 3-hydroxybutyrate (3HB) and 94 mol% 3-hydroxyvalerate (3HV) was produced with a PHA content of as much as 36% of the cellular dry matter. This strain produced PHAs consisting both of the short-chain-length (SCL) and the medium-chain-length (MCL) 3-hydroxyacid units when heptanoic acid to undecanoic acid were fed as the sole carbon sources. Most interestingly, polyester consisting of significant amount of relevant fractions, 3HB, 3HV, and 3-hydroxyheptanoate (3HHp), was accumulated from heptanoic acid. According to solvent fractionation experiments, the polymer produced from heptanoic acid was a blend of poly(3HHp) and of a copolyester of 3HB, 3HV, and 3HHp units. The hexane soluble fractions contained only 3HHp units while the hexane-insoluble fractions contained 3HB and 3HV units with a small amount of 3HHp unit. The copolyester was an elastomer with unusual mechanical properties. The maximum elongation ratio of the copolyester was 460% with an ultimate strength of 10 MPa, which was very different from those of poly(3HB-co-3HV) copolyesters having similar compositions produced from other microorganisms.
This study was aimed to compare the relative cytotoxicity of the five common orthodontic bonding materials (Concise. Mon-lok, Ortho-One, Super C, Transbond) using cell culture technique. DNA synthesis of the fibroblasts was assessed by $^3H$-thymidine uptake to evaluate the effect of the bonding materials on the growth of the cells. The human gingival fibroblasts were explanted from the buccal gingiva of 10 year-old girl and cultured in $\alpha$-MEM/10% FBS/1% antibiotics medium, $37^{\circ}C$, 5% $CO_2$ incuvator. The gingival fibroblasts were tested with the medium into which the bonding materials had been soaked for 1 week. Or the bonding materials were placed on the cells immediately or 2 weeks after polymerization. After 22 hours, $^3H$-thymidine was added into the microtest wells and after 24 hours, the uptake of $^3H$-thymidine was determined by liquid scintilation counter. The results of this study were as follows. 1. DNA synthesis was significantly decreased with Super C and Transbond than Ortho-One, when treated with medium into which the bonding materials had been soaked for 1 week. 2. DNA synthesis was significantly decreased with Concise, Super C and Transbond than control, when treated immediately after polymerization. 3. DNA syntehsis was significantly decreased with Concise, Super C and Transbond than Ortho-One, when immediately after polymerization. 4. There was no significant difference in DNA synthesis between the bonding materials, when treated 2 weeks after polymerization.
Most angiogenesis assays are performed using endothelial cells. However, blood vessels are composed of two cell types: endothelial cells and pericytes. Thus, co-culture of two vascular cells should be employed to evaluate angiogenic properties. Here, we developed an in vitro 3-dimensional angiogenesis assay system using spheroids formed by two human vascular precursors: endothelial colony forming cells (ECFCs) and mesenchymal stem cells (MSCs). ECFCs, MSCs, or ECFCs+MSCs were cultured to form spheroids. Sprout formation from each spheroid was observed for 24 h by real-time cell recorder. Sprout number and length were higher in ECFC+MSC spheroids than ECFC-only spheroids. No sprouts were observed in MSC-only spheroids. Sprout formation by ECFC spheroids was increased by treatment with vascular endothelial growth factor (VEGF) or combination of VEGF and fibroblast growth factor-2 (FGF-2). Interestingly, there was no further increase in sprout formation by ECFC+MSC spheroids in response to VEGF or VEGF+FGF-2, suggesting that MSCs stimulate sprout formation by ECFCs. Immuno-fluorescent labeling technique revealed that MSCs surrounded ECFC-mediated sprout structures. We tested vatalanib, VEGF inhibitor, using ECFC and ECFC+MSC spheroids. Vatalanib significantly inhibited sprout formation in both spheroids. Of note, the $IC_{50}$ of vatalanib in ECFC+MSC spheroids at 24 h was $4.0{\pm}0.40{\mu}M$, which are more correlated with the data of previous animal studies when compared with ECFC spheroids ($0.2{\pm}0.03{\mu}M$). These results suggest that ECFC+MSC spheroids generate physiologically relevant sprout structures composed of two types of vascular cells, and will be an effective pre-clinical in vitro assay model to evaluate pro- or anti-angiogenic property.
In this study, PZT-5A green sheet were prepared by using tape casting technique, and the piezoelectric properties of PZT-5A by variation of sintering temperature was investigated. After, design and piezoelectric properties of 2-2 piezocomposite ultrasonic transducers by menas of the FEA. The acoustic impedance and piezoelectric charge constant of the 2-2 type piezocomposite transducer decreased proportionally due to the density decrease caused by the PZT volume fraction decrease. The piezocomposite acoustic impedance were 7~3 MRayl between 0.6 and 0.2 allowing it to be used for a ultrasonic transducer. The resonance characteristics and the electro-mechanical coupling factor were the best when the volume fraction PZT was 0.6. The PZT volume fraction shows the fixed value, 0.6~0.65, approximately within the range between 0.2 and 0.6 while it is increased to decreased over the range. The result of the experiment above confirmed that the 2-2 piezoelectric composites could be used as the ultrasonic transducers.
For the production and purification of a single chain human insulin precursor, four types of fusion peptides $\beta$-galactosidase (LacZ), maltose binding protein (MBP), glutathione-S-transferase (GST), and (His)(sub)6-tagged sequence (HTS) were investigated. Recombinant E. coli harboring hybrid genes was cultivated at 37$\^{C}$ for 1h, and gene induction occurred when 0.2mM of isopropyl-D-thiogalactoside (IPTG) was added to the culture broth, except for E. coli BL21 (DE3) pLysS harboring a pET-BA cultivation with 1.0mM IPTG, followed by a longer than 4h batch fermentation respectively. DEAE-Sphacel and Sephadex G-200 gel filtration chromatography, amylose affinity chromatography, glutathione-sepharose 4B affinity chromatography, and a nickel chelating affinity chromatography system as a kind of immobilized metal ion affinity chromatography (IMAC) were all employed for the purification of a single chain human insulin precursor. The recovery yields of the HTS-fused, GST-fused, MBP-fused, and LacZ-fused single chain human insulin precursors resulted in 47%, 20%, 20%, and 18% as the total protein amounts respectively. These results show that a higher recovery yield of the finally purified recombinant peptides was achieved when affinity column chromatography was employed and when the fused peptide had a smaller molecular weight. In addition the pET expression system gave the highest productivity of a fused insulin precursor due to a two-step regulation of the gene expression, and the HTS-fused system provided the highest recovery of a fused insulin precursor based on a simple and specific separation using the IMAC technique.
The incorporation of Shiitake culture into sawdust is a widely utilized technique that can assist in reducing the cost and time consumption associated with oak cultivation. In sawdust cultivation, browning of the surface mycelia is an important stage with respect to the utility and longevity of the sawdust media. Surface browning forms a protective coating on the substrate, which can inhibit the invasion of pathogens and suppress water evaporation. Several different light sources (red LED, white LED, blue LED, and fluorescent light) were used and the intensity of illumination was carefully controlled (1.5, 10.5, $20.5{\mu}mol/m^2s$ for LEDs and 10, 100, 300 lux for the fluorescent light) to induce browning. The light sources were regulated via a 1 h on/off cycle in a controlled room environment at a temperature of $20^{\circ}C$, 60% humidity, and 1200 ppm $CO_2$ concentration for 60days. The browning effect varied depending on the source and the intensity of illumination. This effect was most effectively induced at $1.5{\mu}mol/m^2s$ for the red and blue LEDs. All light sources induced less browning at the highest intensity of illumination. This indicates that intensity values higher than $20.5{\mu}mol/m^2s$ in the case of the LEDs and 300 lux for the fluorescent light are not effective. After harvesting of the fruit bodies, we measured the weight, length, and width of the pileus and stipe in addition to their chromaticity and hardness. Treatment with $1.5{\mu}mol/m^2s$ blue LED produced the best harvest with the highest average chromaticity, weight (21.2 g), stipe length (30.8 mm), and hardness (377.9 g), with a fine length and width of the pileus.
The research was conducted(1) to confirm the agent(s) responsible for the antimicrobial activity contained in the fermented tomato juice with L. acidophilus(2) to extract and purify the antimicrobial agent(s)(3) to find the biological, physical and chemical properties of the agent(s). The following results were obtained and summarized as followings; 1. The agent responsible for the inhibitory activity was confirmed by both well assay method using fermented tomato juice with L. acidophilus and turbidimetric technique using the cell-free filtrate or neutralized filtrate of tomato acidohilus culture and found exerted antimicrobial agent other than lactic acid. 2. The procedures of purification : The isolation and purification of antimicrobial agent from the lyophilized acidophilus tomato culture were carried out by (1) methanol extraction (2) acetone extraction, (3) Sephadex G-50 gel filtration (4) paper chromatography and (5) thin layer chromatography. 3. The biological, physical and chemical properties of antimicrobial agent: The biological, physical, chemical properties of the purified antimicrobial agent were: (1) The antimicrobial activity was strong against test organisms; Bacillus subtilis(ATCC 6633), Escheichia coli(ATCC 25922), Staphylococcus aureus(ATCC 167), Pseudomonas fluorescens(KFCC 32394), Proteus vulgaris and Shigella dysenteriae. (2) The pH value of the agent was 2.0 and the inhibitory activity was lost when it was neutralized at 7.0 of pH and the agent was heat stable at $121^{\circ}C$ for 60 minutes. (3) The ultraviolet light absorption spectra of methanol-acetone extract and TLC fraction exhibited a maximum absorption at 260nm and 224nm respectively. (4) The most purified agent from TLC plate increased about 130-fold in activity. (5) The agent isolated from TLC plate was free from $H_2O_2$ or lactic acid. 4. Bioautographic assy: By means of bioautography of the agent on silica gel of TLC plate a strong inhibitory activity against B. subtilis was demonstrated. 5. Mass spectrometry: The agent obtained from TLC plate was analyzed by mass spectrometry which show the parent peak at m/e 264 suggesting the molecular weight of the compound and molecular group such as [$C_2H{^+}_4$], [CO], [CH=NH], [$C_3{H^}4_7$], [$\begin{array}{rcl}O\\{\parallel}\\CH_3-C\\\end{array}$], [$C_6-H{^+}_{11}$], [$C_5H{^+}_{11}$], [$\begin{array}{rcl}O\\{\parallel}\\C_5H_7-C^+\\\end{array}$] were suggested.
A scaling up study for the exopolysaccharide (EPS) production by submerged culture of Ganoderma lucidum was carried out in jar fermenter systems (2.6, 20 and 75 L) under bi-staged pH process. Profiles of dissolved oxygen (DO) and volumetric coefficient of oxygen transfer ($k_La$) as a function of operating variables (agitation speed and aeration rate) was investigated, and a correlation between $k_La$ and operating variables was analysed statistically. Under bi-staged pH process, no limitation of DO was observed at agitation speeds tested in the range of 200 and 600 rpm, and the highest EPS production was obtained at the level of DO of $40{\sim}80%$. From the regression analysis, the relation between $k_La$, gas velocity (Vs), stirrer speed (N) and impeller diameter (Di) could be expressed as : $$k_La=0.555{\times}Vs^{0.42}{\times}(N^3{\times}Di^2)^{0.33}\;(R^2=0.925,\;p<0.05)$$ It was found that under 2.6 L jar fermenter, the optimum agitation speed and aeration rate was 400 rpm and 1 vvm, respectively, obtaining the EPS production of 15.43 g/L. Under the submerged cultivation of G. lucidum in jar fermenters of $2.6{\sim}75\;L$, the similar EPS yields at each fermenter were achieved during scaling up based on $k_La$, and $k_La$ value for maximum EPS production was $85.4{\pm}26.70\;h^{-1}$.
Porphyromonas endodontalis, an anaerobic Gram negative cocobacillus which was known to be associated with the infected root canals and periapical lesions, is very difficult to culture and to detect by the traditional method in that it requires much time to induce the specific black pigmentation, and it is very sensitive to oxygen and the antibiotics added in the culture medium. In this study, the nucleotide sequences of the 'probe h' (0.73kb), one of the specific DNA probes top. endodontalis (ATCC 35406) which had been developed by our department, was determined and then a pair of primers for PCR amplification was fabricated to identify P. endodontalis. The plasmids containing 'probe h' were purified by $Wizard^{TM}$ Midipreps DNA Purification System (Promega Corp.), and the nucleotide sequences of the 'probe h' were determined by the dideoxy chain termination method using TaqTrack Sequencing System (Promega Corp.) and detected by fluorescent labelling method. The sense/antisense PCR primers were designed with computer software (Lasergene, DNASTAR Ind. PCR was done with a programmable GeneAmp PCR System 2400 (Perkin Elmer-Cetus Co.). Each sample containing the whole genomic DNA of P. endodontalis and other black-pigmented Bacteroides was itailly denatured at $94^{\circ}C$ for 5 min and then subjected to 30 cycles, each of them consisting of 60s at $94^{\circ}C$, 60s at $60^{\circ}C$, and 90s. at $72^{\circ}C$. The amplified DNA was resolved electrophoretically in a 1.0 % agarose gel in 1X TAE buffer, stained with EtBr, and photographed on a UV transilluminator. The results were as follows : 1. The nucleotide sequences of 'probe h' (743 base pairs) were obtained by dideoxy chain termination method, and from that results the specific primers to P. endodontalis (ATCC 35406), 'Primer H1/ Primer H2', were designed. 2. It has been found that 'Primer H1/H2' could detect P. endodontalis (ATCC 35406) using PCR. 3. The PCR system with this primers may be a powerful technique to amplify the specific sequences of 'probe h' of P. endodontalis (ATCC 35406) that produce distinct identification of it from other black-pigmented Bacteroides, and this could help us to determine the nature of periapical disease.
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