• The Korean Journal of Food And Nutrition
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• v.23 no.4
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• pp.556-561
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• 2010

Tomato fruits(Lycoperisicon esculentum) synthesize the glycoalkaloids dehydrotomatine and ${\alpha}$-tomatine, possibly as defense against bacteria, fungi and insects. We developed a new effective method to prepare and purify dehydrotomatine and ${\alpha}$-tomatine that exists in tomato fruits using alumina column chromatography and high performance liquid chromatography (HPLC). The tomato glycoalkaloids(TGA) in tomato was extracted with 2% acetic acid, and then precipitated with ammonium hydroxide(pH=10.5). The dry precipitate substance was applied on alumina column, and then fractionated with water saturated n-butylalcohol. The TGA(Fr. No. 26~36) were collected and dried under reduced pressure. The TGA was performed on a reverse phase HPLC(Inertsil ODS-2, $5\;{\mu}m$), eluted with acetonitrile/20mM $KH_2PO_4$(24:76, v/v) at 208 nm. Two peaks were detected on HPLC, and individual peak was collected by repeating HPLC. Furthermore, to confirm the identity dehydrotomatine and ${\alpha}$-tomatine, each peak isolated was hydrolyzed with 1N HCl into sugar and aglycone tomatidine. The sugars were converted to trimethylsilyl ester derivatives. The nature and molar ratios of sugars were identified by gas-liquid chromatography(GLC) and the aglycone by high-performance liquid chromatography(HPLC). The first peak (Rt=17.5 min) eluted from HPLC was identified as dehydrotomatine, and second peak(Rt=21.0 min) was as ${\alpha}$-tomatine. This technique has been used effectively to prepare and isolate dehydrotomatine and ${\alpha}$-tomatine from tomato fruits.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.30 no.5
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• pp.438-443
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• 2004

Objectives : The proper development of the facial structures relies upon a sequence of tightly regulated signaling interactions between the ectoderm and mesoderm involving the participation of several families of signaling molecules. Among these, bone morphogenetic proteins (BMPs) have been suggested to be a key signal that regulates the development of the mandible and the initiation and morphogenesis of the teeth. The aim of this study was to examine the artificial development of the mandibular structures and to examine the role of BMPs on tooth morphogenesis and differentiation using an organ culture system. Materials and Methods : The tooth germs from Ed 11.5, 13.5 mice were dissected, and transplanted into the diastema of the mandible primordia. The mandibles containing the transplanted tooth germs were cultured in vitro. During this period, beads soaked with BMP4 were implanted around the transplanted tooth germs. In addition, a diastema block containing the transplanted tooth germ was dissected, then transferred to an adult mouse kidney. After the organ culture, the developing mandibular explant was removed from the kidney and prepared for the tissue specimens. Odontogeneis of the transplanted tooth germs was examined after Hematoxylin-eosin, Masson-trichrome staining. Results : Proliferation and differentiation of the tooth germs cultured in the diastema was observed. In the BMP4-treated tooth germs, the formation of the first and second molars was noted. The crown of the developing tooth showed the formation of a mature cusp with the deposition of enamel and dentin matrix. In conclusion, it was confirmed that BMP4 is involved in the formation of a dental crown and the differentiation of ameloblasts and odontoblasts of the molar tooth during the development of the transplanted tooth germs.

• Nutritional Sciences
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• v.5 no.4
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• pp.175-183
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• 2002

To compare the effects of various types of dietary fiber on microbial production of short-chain fatty acids (SCFA) and on colon cell proliferation which is used as an intermediate biomarker for colon carcinogenesis, groups of 10 male Sprague-Dawley rats were fed one of four fiber-supplemented diets (6% cellulose, 6% pectin, 6% polydextrose, and a mixture of 3% cellulose and 3% polydextrose) for three weeks. As a control, a fiber-free diet was fed to a separate group of 10 rats. Cell proliferation was measured by in vivo incorporation of bromodeoxyuridine into DNA in the proximal and distal colon, respectively. Luminal concentrations of SCFA were measured by gas chromatography. Dietary fiber significantly influenced microbial production of SCFA in the colon; pectin supplementation produced the highest concentrations of luminal SCFA in both the proximal and distal colon (p<0.05). The degree of individual SCFA production was characterized by a relatively higher increase in butyrate production by the pectin-supplemented diet, and in propionate production by the polydextrose-supplemented diet, resulting in alterations of the molar ratios of acetate, propionate and butyrate. There were significant differences in colon cell proliferation among the diet groups; the pectin-supplemented diet produced a significantly higher effect on cell proliferation of distal colonic epithelial cells (p＜0.05), and the polydextrose-supplemented diet produced an intermediate effect compared to the fiber-free or cellulose-supplemented diet. Increased cell proliferation was correlated to increased luminal concentrations of butyrate in the proximal colon and to increased luminal concentrations of propionate and butyrate in the distal colon (p<0.05). Therefore, these data suggest that dietary fiber may modulate colon cell proliferation by altering luminal SCFA concentrations, particularly butyrate and perhaps propionate. In addition, the present study is the first finding that has demonstrated a relative increase in colon cell proliferation due to supplementation with polydextrose, suggesting that the overuse of this artificially synthesized polysaccharide in food processing technology needs to be carefully evaluated from the public health point of view.

• Microbiology and Biotechnology Letters
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• v.46 no.1
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• pp.9-17
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• 2018

The aims of this work were to characterize and determine bioactivities of crude exopolysaccharide (EPS) extract from Bacillus tequilensis PS21 isolated from milk kefir from Kampaeng Petch, Thailand. B. tequilensis PS21 produced 112.1 mg dried EPS/l from initial 80 g/l lactose in modified TSB media at 52 h, with EPS product yield of 8.9 mg EPS/g lactose and specific product yield of 0.3 mg EPS/mg biomass. The FTIR result confirmed EPS to be a protein-bound polysaccharide and SEM analysis showed the morphology to be a grainy appearance with an uneven surface, covered with pores. HPLC analysis determined EPS as a heteropolysaccharide consisting of five sugar units with the following molar ratios; xylose (17.65), glucose (2.54), ribose (1.83), rhamnose (1.23), and galactose (1). Chemical components of this EPS were predominantly carbohydrate at 697.8 mg/g EPS (65%), protein 361.4 mg/g EPS (34%), and nucleic acid 12.5 mg/g EPS (1%). The EPS demonstrated antioxidant activities at 57.5% DPPH scavenging activity, $37.2{\mu}M\;Fe(II)/mg$ EPS and $34.9{\mu}M\;TEAC\;{\mu}M/mg$ EPS using DPPH, FRAP and ABTS assays, respectively. EPS also exhibited anti-tyrosinase activity at 34.9% inhibition. This work represents the first finding of EPS produced by Bacillus sp. from Thai milk kefir which shows potential applications in the production of antioxidant functional foods and whitening cosmetics. However, optimization of EPS production for industrial exploitation requires further study to ascertain the economic potential.

• Journal of Microbiology and Biotechnology
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• v.29 no.12
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• pp.1882-1893
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• 2019

β-Glucosidases and β-xylosidases are two categories of enzymes that could cleave out non-reducing, terminal β-D-glucosyl and β-D-xylosyl residues with release of D-glucose and D-xylose, respectively. In this paper, two functional β-glucosidase Dth3 and β-xylosidase Xln-DT from Dictyoglomus thermophilum were heterologously expressed in E.coli BL21 (DE3). Dth3 and Xln-DT were relatively stable at 75℃ and were tolerant or even stimulated by glucose and xylose. Dth3 was highly tolerant to glucose with a K_i value of approximately 3 M. Meanwhile, it was not affected by xylose in high concentration. The activity of Xln-DT was stimulated 2.13-fold by 1 M glucose and 1.29-fold by 0.3 M xylose, respectively. Furthermore, the βglucosidase Dth3 and β-xylosidase Xln-DT showed excellent selectivity to cleave the outer C-6 and C-3 sugar moieties of ASI, which established an effective and green method to produce the more pharmacologically active CAG, an exclusive telomerase activator. We measured temperature, pH and dosage of enzyme using a single-factor experiment in ASI biotransformation. After optimization, the optimal reaction conditions were as follows: 75℃, pH 5.5, 1 U of Dth3 and 0.2 U of Xln-DT, respectively. Under the optimized conditions, 1 g/l ASI was transformed into 0.63 g/l CAG with a corresponding molar conversion of 94.5% within 3 h. This is the first report to use the purified thermostable and sugar-tolerant enzymes from Dictyoglomus thermophilum to hydrolyze ASI synergistically, which provides a specific, environment-friendly and cost-effective way to produce CAG.

• Journal of Microbiology and Biotechnology
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• v.28 no.2
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• pp.255-261
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• 2018

Aglycon protopanaxatriol (APPT) has valuable pharmacological effects such as memory enhancement and tumor inhibition. ${\beta}$-Glycosidase from the hyperthermophilic bacterium Dictyoglomus turgidum (DT-bgl) hydrolyzes the glucose residues linked to APPT, but not other glycoside residues. ${\beta}$-Glycosidase from the hyperthermophilic bacterium Pyrococcus furiosus (PF-bgl) hydrolyzes the outer sugar at C-6 but not the inner glucose at C-6 or the glucose at C-20. Thus, the combined use of DT-bgl and PF-bgl is expected to increase the biotransformation of PPT-type ginsenosides to APPT. We optimized the ratio of PF-bgl to DT-bgl, the concentrations of substrate and enzyme, and the reaction time to increase the biotransformation of ginsenoside Re and PPT-type ginsenosides in Panax ginseng leaf extract to APPT. DT-bgl combined with PF-bgl converted 1.0 mg/ml PPT-type ginsenosides in ginseng leaf extract to 0.58 mg/ml APPT without other ginsenosides, with a molar conversion of 100%. We achieved the complete biotransformation of ginsenoside Re and PPT-type ginsenosides in ginseng leaf extract to APPT by the combined use of two ${\beta}$-glycosidases, suggesting that discarded ginseng leaves can be used as a source of the valuable ginsenoside APPT. To the best of our knowledge, this is the first quantitative production of APPT using ginsenoside Re, and we report the highest concentration and productivity of APPT from ginseng extract to date.

• Analytical Science and Technology
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• v.23 no.1
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• pp.60-67
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• 2010

A sensitive competitive heterogeneous bioluminescence immunoassay for serotonin was developed using photoprotein, aequorin as a label for the first time with the optimal assay conditions; especially, serotoninavidin conjugate was prepared by Mannich reaction and the synthetic process of serotonin-avidin conjugate was optimized by controlling the initial molar ratios of serotonin, formaldehyde and avidin (1:12,000:25). The developed bioluminescence immunoassay for serotonin showed good sensitivity (LOD of 0.68 ng/mL) with wide area of dynamic range ($5.0{\times}10^{-10}\;M\sim5.0{\times}10^{-7}\;M$). (cf. the range for serotonin in human blood serum is $151{\pm}45\;ng$/mL). In addition, cross-reactivity studies demonstrated that 5-methoxytryptamine showed some cross-reactivity (28.0%), whereas 3-methylindole, melatonin and 5-hydroxylindole-3-acetic acid showed no crossreactivity, and good recoveries were obtained in serum. Thus, this developed method provides a good tool to monitor serotonin in serum.

• Advances in concrete construction
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• v.2 no.2
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• pp.109-123
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• 2014

This paper presents chloride induced corrosion durability of reinforcing steel in geopolymer concretes containing different contents of sodium silicate ($Na_2SiO_3$) and molarities of NaOH solutions. Seven series of mixes are considered in this study. The first series is ordinary Portland cement (OPC) concrete and is considered as the control mix. The rest six series are geopolymer concretes containing 14 and 16 molar NaOH and $Na_2SiO_3$ to NaOH ratios of 2.5, 3.0 and 3.5. In each series three lollypop specimens of 100 mm in diameter and 200 mm in length, each having one 12 mm diameter steel bar are considered for chloride induced corrosion study. The specimens are subjected to cyclic wetting and drying regime for two months. In wet cycle the specimens are immersed in water containing 3.5% (by wt.) NaCl salt for 4 days, while in dry cycle the specimens are placed in open air for three days. The corrosion activity is monitored by measuring the copper/copper sulphate ($Cu/CuSO_4$) half-cell potential according to ASTM C-876. The chloride penetration depth and sorptivity of all seven concretes are also measured. Results show that the geopolymer concretes exhibited better corrosion resistance than OPC concrete. The higher the amount of $Na_2SiO_3$ and higher the concentration of NaOH solutions the better the corrosion resistance of geopolymer concrete is. Similar behaviour is also observed in sorptivity and chloride penetration depth measurements. Generally, the geopolymer concretes exhibited lower sorptivity and chloride penetration depth than that of OPC concrete. Correlation between the sorptivity and the chloride penetration of geopolymer concretes is established. Correlations are also established between 28 days compressive strength and sorptivity and between 28 days compressive strength and chloride penetration of geopolymer concretes.

• Environmental Engineering Research
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• v.20 no.1
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• pp.89-97
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• 2015

Remediation of wastewater contaminated with metal(II)-complexed species (Cu(II)-NTA (NTA: nitrilotriacetic acid), Cu(II)-EDTA (EDTA: ethylenediamine tetraacetic acid) and Cd(II)-EDTA is attempted using the potential applicability of ferrate(VI). Kinetics of pollutant degradation is obtained with the removal of ferrate(VI) studied at wide range of pH (8.0-10.0) and the concentration of metal(II)-complexed species (0.3 to 15.0 mmol/L) employing a constant dose of ferrate(VI) i.e., 1.0 mmol/L. Pseudo-first-order and pseudo-second-order rate constants were obtained in the reduction of ferrate(VI) which was then employed to obtain the overall rate constants of the pollutant degradation. The mineralization of NTA and EDTA was obtained with the change in TOC (total organic carbon) values collected by the ferrate(VI) treated pollutant samples. Decrease in pH and molar pollutant concentrations was greatly favored the percent mineralization of NTA or EDTA by the ferrate(VI) treatment. The treated pollutant samples were filtered and subjected for AAS (atomic absorption spectrophotometric) analysis to assess the simultaneous removal of copper and cadmium from aqueous solutions at the studied pH as well at the elevated pH 12.0. Results show that an enhanced removal of cadmium or copper was achieved at pH 12.0. Overall, ferrate(VI) possesses multifunctional application in wastewater treatment as it oxidizes the degradable impurities and removes metallic impurities by coagulation process.

• Environmental Engineering Research
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• v.22 no.1
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• pp.19-30
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• 2017

Direct catalysis of vapors from vacuum pyrolysis of biomass was performed on MCM-41 to investigate the effects of operating parameters including catalyzing temperature, catalyzing bed height and system pressure on the organic yields. Optimization of organic phase yield was further conducted by employing response surface methodology. The statistical analysis showed that operating parameters have significant effects on the organic phase yield. The organic phase yield first increases and then decreases as catalyzing temperature and catalyzing bed height increase, and decreases as system pressure increases. The optimal conditions for the maximum organic phase yield were obtained at catalyzing temperature of $502.7^{\circ}C$, catalyzing bed height of 2.74 cm and system pressure of 6.83 kPa, the organic phase yield amounts to 15.84% which is quite close to the predicted value 16.19%. The H/C, O/C molar ratios (dry basis), density, pH value, kinematic viscosity and high heat value of the organic phase obtained at optimal conditions were 1.287, 0.174, $0.98g/cm^3$, 5.12, $5.87mm^2/s$ and 33.08 MJ/kg, respectively. Organic product compositions were examined using gas chromatography/mass spectrometry and the analysis showed that the content of oxygenated aromatics in organic phase had decreased and hydrocarbons had increased, and the hydrocarbons in organic phase were mainly aliphatic hydrocarbons. Besides, thermo-gravimetric analysis of the MCM-41 zeolite was conducted within air atmosphere and the results showed that when the catalyst continuously works over 100 min, the index of physicochemical properties of bio-oil decreases gradually from 1.15 to 0.45, suggesting that the refined bio-oil significantly deteriorates. Meanwhile, the coke deposition of catalyst increases from 4.97% to 14.81%, which suggests that the catalytic activity significantly decreases till the catalyst completely looses its activity.