• Journal of Life Science
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• v.15 no.6 s.73
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• pp.851-856
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• 2005

Although valuable microbes have been isolated from the soil for the various productions of useful components, the microbes which can be cultivated in the laboratory are only $0.1-1\%$ of all microbes. To solve this problem, the study has recently been tried for making the valuable components from the environment by directly separating unculturable micrbial DNA in the soil. But it is known that humic acid originated from the soil interrupts various restriction enzymes and molecular biological process. Thus, in order to prevent these problems, this study modified the method separated soil DNA with phenol, CTAB and PEG. In order to compare the degree of purity for each DNA and the molecular biological application process, $A_{260}/A_{280}$ ratio, restriction enzymes, and PCR were performed. In case of DNA by the modified method, total yield of DNA was lower but $A_{260}/A_{280}$ ratio was higher than the previously reported methods. It was confirmed that the degree of purity is improved by the modified method. But it was not cut off by all kinds of tested restriction enzymes because of the operation of a very small amount of interrupting substances. When PCR was operated with each diluted DNA in different concentrations and GAPDH primer, the DNA by the modified method could be processed for PCR in the concentration of 100 times higher than by the previously reported separation method. Therefore, this experiment can find out the possibility of utilization for the unknown substances by effectively removing the harmful materials including humic acid and help establishing metagenomic DNA library from the soil DNA having the high degree of purity.

• Research in Plant Disease
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• v.30 no.3
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• pp.294-299
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• 2024

Polymerase chain reaction (PCR) methods, including conventional PCR (cPCR) and quantitative real-time PCR (qRT-PCR), with both plasmid- and chromosome-targeting primers, are currently the most reliable methods for detecting Erwinia amylovora due to their high sensitivity and specificity. Despite qRT-PCR's quantitative advantage, cPCR remains an attractive method to detect this bacterium in initial screenings of suspected host plants, as it is cost-effective and does not require skilled personnel in well-equipped laboratories. This study aimed to significantly improve cPCR robustness via application of bovine serum albumin (BSA) as a PCR facilitator, with a modified EaF/R primer pair, as previously reported. Experiments have shown that simple supplementation with BSA (10 mg/ml) enhances cPCR reactions using templates such as genomic DNA, bacterial cells, and infected symptomless host organs, including immature apple fruits and seedlings, with EaF/R primers. The cPCR method described in this study is simple, specific, and reliable, and can be applied in routine assays to diagnose fire blight.

• Journal of Dental Rehabilitation and Applied Science
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• v.29 no.1
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• pp.37-44
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• 2013

Sodium hypochlorite and ethylene diamine tetra acetic acid are substances usually used during endodontic treatment. Several studies found that the bonding was negated with certain irrigants and some of the used irrigants have demineralizing and chealating effects, so it was advocated to omit the etching step in etch and rinse adhesive systems. The purpose of this in vitro study was to evaluate the influence of NaOCl & EDTA on the bonding strength of ethanol wet bonding. Thirty human molars were selected and mesiodistally sectioned into halves, thus providing sixty specimens. The specimens were randomly assigned to 4 groups(n=15) according to the irrigant regimen used : (1) irrigated with distilled water for 10min (control); (2) irrigated with 5.25% NaOCl(10min), flushed with 5.25% NaOCl(1min) (3) irrigated with 5.25% NaOCl, flushed with 17% EDTA (4) irrigated with 5.25% NaOCl, flushed with 17% EDTA. Each group was acid-etched with 37% phosphoric acid(except group 4) and had their dentin surfaces dehydrated with ethanol solutions : 50%, 70%, 80%, 95%, 3x100%, 30s for each application. After dehydration, a primer( 50% all bond 3 resin + 50% ethanol) was used, followed by the adhesive(ALL-BOND 3 RESIN) application. Resin composite build-ups were then prepared using an incremental technique. Specimens were sectioned into beams and submitted to a tensile load using a Micro Tensile Tester(Bisco Inc.). The data were statistically analyzed using one-way ANOVA and Tukey HSD at p<0.5 level. There was no significant difference on G1(control) and G2(irrigated with NaOCl only ). (p>0.05). G3(flushed with EDTA) showed significantly high tensile bonding strength compared to the G2 (p<0.05). G4( treated with EDTA but no acid-etching) was significantly lower value than G3. (p<0.05) Although there was no significant difference, 5.25% NaOCl seemed to have an adverse effect on the bonding strength of ethanol wet bonding. The flushing with EDTA after NaOCl irrigation prevents the decrease of bonding strength. The use of 17% EDTA as a final flush can enhance the bonding strength but EDTA flushing can't substitute for a acid-etching.

• Restorative Dentistry and Endodontics
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• v.27 no.2
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• pp.150-157
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• 2002

One of the latest concepts in bonding are "total etch", in which both enamel and dentin are etched with an acid to remove the smear layers, and "wet dentin" in which the dentin is not dry but left moist before application of the bonding primer Ideally the application of a bonding agent to tooth structure should be insensitive to minor contamination from oral fluids. Clinically, contaminations such as saliva, gingival fluid, blood and handpiece lubricant are often encountered by dentists during cavity preparation. The aim of this study was to evaluate the effect of contamination by hemostatic agents on shear bond strength of compomer restorations. One hundred and ten extracted human maxillary and mandibular molar teeth were collected. The teeth were removed soft tissue remnant and debris and stored in physiologic solution until they were used. Small flat area on dentin of the buccal surface were wet ground serially with 400, 800 and 1200 abrasive papers on automatic polishing machine. The teeth were randomly divided into 11 groups. Each group was conditioned as follows : Group 1: Dentin surface was not etched and not contaminated by hemostatic agents. Group 2: Dentin surface was not etched but was contaminated by Astringedent$^{\circledR}$(Ultradent product Inc., Utah, U.S.A.) Group 3: Dentin surface was not etched but was contaminated by Bosmin$^{\circledR}$(Jeil Pharm, Korea.). Group 4: Dentin surface was not etched but was contaminated by Epri-dent$^{\circledR}$(Epr Industries, NJ, U.S.A.). Group 5: Dentin surface was etched and not contaminated by hemostatic agents. Group 6: Dentin sorface was etched and contaminated by Astringedent$^{\circledR}$. Group 7 : Dentin surface was etched and contaminated by Bosmin$^{\circledR}$. Group 8: Dentin surface was etched and contaminated by Epri-dent$^{\circledR}$. Group 9: Dentin surface was contaminated by Astringedent$^{\circledR}$. The contaminated surface was rinsed by water and dried by compressed air. Group 10: Dentin surface was contaminated by Bosmin$^{\circledR}$. The contaminated surface was rinsed by water and dried by compressed air. Group 11 : Dentin surface was contaminated by Epri-dent$^{\circledR}$. The contaminated surface was rinsed by water and dried by compressed air. After surface conditioning, F2000$^{\circledR}$ was applicated on the conditoned dentin surface The teeth were thermocycled in distilled water at 5$^{\circ}C$ and 55$^{\circ}C$ for 1,000 cycles. The samples were placed on the binder with the bonded compomer-dentin interface parallel to the knife-edge shearing rod of the Universal Testing Machine(Zwick Z020, Zwick Co., Germany) running at a cross head speed or 1.0 mm/min. Group 2 showed significant decrease in shear bond strength compared with group 1 and group 6 showed significant decrease in shear bond strength compared with group 5. There were no significant differences in shear bond strength between group 5 and group 9, 10 and 11.

• Restorative Dentistry and Endodontics
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• v.24 no.4
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• pp.647-656
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• 1999

The purpose of this study was to investigate the shear bond strength and marginal microleakage of composite to enamel and dentin according to different treatment methods when the applied bonding agent was contaminated by artificial saliva. For the shear bond strength test, the buccal and occlusal surfaces of one hundred twenty molar teeth were ground to expose enamel(n=60) and dentin surfaces(n=60). The specimens were randomly assigned into control and 5 experimental groups with 10 samples in each group. In control group, a bonding system(Scotchbond$^{TM}$ Multi-Purpose plus) and a composite resin(Z-100$^{TM}$) was bonded on the specimens according to manufacture's directions. Experimental groups were subdivided into 5 groups. After polymerization of an adhesive, they were contaminated with at artificial saliva on enamel and dentin surfaces: Experimental group 1 ; artificial saliva was dried with compressed air. Experimental group 2 ; artificial saliva was rinsed with air-water spray and dried. Experimental group 3 ; artificial saliva was rinsed, dried and applied an adhesive. Experimental group 4 ; artificial saliva was rinsed, dried, and then etched using phosphoric acid followed by an adhesive. Experimental group 5, artificial saliva was rinsed, dried, and then etched with phosphoric acid followed by consecutive application of both a primer and an adhesive. Composite resin(Z-100$^{TM}$) was bonded on saliva-treated enamel and dentin surfaces. The shear bond strengths were measured by universal testing machine(AGS-1000 4D, Shimaduzu Co. Japan) with a crosshead speed of 5mm/minute under 50kg load cell. Failure modes of fracture sites were examined under stereomicroscope. The data were analyzed by one-way ANOVA and Tukey's test. For the marginal microleakage test, Class V cavities were prepared on the buccal surfaces of sixty molars. The specimens were divided into control and experimental groups. Cavities in experimental group were contaminated with artificial saliva and those surfaces in each experimental groups received the same treatments as for the shear test. Cavities were filled with Z-100. Specimens were immersed in 0.5% basic fuchsin dye for 24 hours and embedded in transparent acrylic resin and sectioned buccolingually with diamond wheel saw. Four sections were obtained from the one specimen. Marginal microleakages of enamel and dentin were scored under streomicroscope and averaged from four sections. The data were analyzed by Kruskal-Wallis test and Fisher's LSD. The results of this study were as follows. 1. The shear bond strength to enamel showed lower value in experimental group 1(13.20${\pm}$2.94MPa) and experimental group 2(13.20${\pm}$2.94MPa) than in control(20.03${\pm}$4.47MPa), experimental group 4(20.96${\pm}$4.25MPa) and experimental group 5(21.25${\pm}$4.48MPa) (p<0.05). 2. The shear bond strength to dentin showed lower value in experimental group 1(9.35${\pm}$4.11MPa) and experimental group 2(9.83${\pm}$4.11MPa) than in control group(17.86${\pm}$4.03MPa), experimental group 4(15.04${\pm}$3.22MPa) and experimental group 5(14.33${\pm}$3.00MPa) (p<0.05). 3. Both on enamel and dentin surfaces, experimental group 1 and 2 showed many adhesive failures, but control and experimental group 3, 4 and 5 showed mixed and cohesive failures. 4. Enamel marginal microleakage was the highest in experimental group 1 and there was a significant difference in comparison with other groups (p<0.05). 5. Dentin marginal microleakages of experimental group 1 and 2 were higher than those of other groups (p<0.05). This result suggests that treatment methods, re-etching with 35% phosphoric acid followed by re-application of adhesive or repeating all adhesive procedures, will produce good effect on both shear bond strength and microleakage of composite to enamel and dentin if the polymerized bonding agent was contaminated by saliva.

• The Journal of Korean Academy of Prosthodontics
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• v.46 no.5
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• pp.535-543
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• 2008

Purpose: The objective of this study was to compare the bonding characteristics of a new self-adhesive resin cement to dentin, which does not require bonding and conditioning procedure of the tooth surface, and conventional resin cement. The effect of phosphoric acid etching prior to application of self-adhesive resin cement on the shear bond strength was also evaluated. Material and methods: Fortyfive non-carious human adult molars extracted within 6 months were embedded in chemically cured acrylic resin. The teeth were ground with a series of SiC-papers ending with 800 grit until the flat dentin surfaces of the teeth were exposed. The teeth were randomly divided into 3 experimental groups. In group 1, self-adhesive resin cement, RelyX Unicem (3M ESPE, Seefeld, Germany) was bonded without any conditioning of teeth. In group 2, RelyX Unicem was bonded to teeth after phosphoric acid etching. For group 3, Syntac Primer (Ivoclar Vivadent AG, Schaan, Liechtenstein) was applied to the teeth before Syntac adhesive (Ivoclar Vivadent AG, Schaan, Liechtenstein) and Helibond (Ivoclar Vivadent AG, Schaan, Liechtenstein) followed by conventional resin cement, Variolink II (Ivoclar Vivadent AG, Schaan, Liechtenstein). To make a shear bond strength test model, a plastic tuble (3 mm diameter, 3 mm height) was applied to the dentin surfaces at a right angle and filled it with respective resin cement, and light-polymerized for 40 seconds. All the specimens were stored in distilled water at $37^{\circ}C$ for 24 hours before test. Universal Testing Machine (Z020, Zwick, Ulm, Germany) at a cross head speed of 1 mm/min was used to evaluate the shear bond strength. The failure sites were inspected under a magnifier and Scanning Electron Microscope. The data was analyzed with One way ANOVA and Scheffe test at ${\alpha}$= 0.05. Results: (1) The shear bond strengths to dentin of RelyX Unicem was not significantly different from those of Variolink II/Syntac. (2) Phosphoric acid etching lowered the shear bond strength of RelyX Unicem significantly. (3) Most of RelyX Unicem and Variolink II showed mixed fractures, while all the specimens of RelyX Unicem with phosphoric acid etching demonstrated adhesive failure between dentin and resin cement. Conclusion: Shear bond strength to dentin of self-adhesive resin cement is not significantly different from conventional resin cement, and phosphoric acid etching decrease the shear bond strength to dentin of self-adhesive resin cement.

• Clinical and Experimental Pediatrics
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• v.50 no.7
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• pp.686-693
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• 2007

Purpose : Mycophenolic acid (MPA), the active metabolite of mycophenolate mofetil (MMF), is a potent inhibitor of inosine-monophosphate dehydrogenase (IMPDH), a new immunosuppressive drug used. It was reported that MPA protected neurons after excitotoxic injury, induced apoptosis in microglial cells. However, the effects of MPA on hypoxic-ischemic (HI) brain injury has not been yet evaluated. Therefore, we examined whether MPA could be neuroprotective in perinatal HI brain injury using Rice-Vannucci model (in vivo) and in rat brain cortical cell culture induced by hypoxia (in vitro). Methods : Cortical cells were cultured using a 18-day-pregnant Sprague-Dawley (SD) rats and incubated in 1% $O_2$ incubator for hypoxia. MPA ($10{\mu}g/mL$) before or after a HI insult was treated. Seven-day-old SD rat pups were subjected to left carotid occlusion followed by 2 hours of hypoxic exposure (8% $O_2$). MPA (10 mg/kg) before or after a HI insult were administrated intraperitoneally. Apoptosis was measured using western blot and real-time PCR for Bcl-2, Bax, caspase-3. Results : H&E stain revealed increased brain volume in the MPA-treated group in vivo animal model of neonatal HI brain injury. Western blot and real-time PCR showed the expression of caspase-3 and Bax/Bcl-2 were decreased in the MPA-treated group In in vitro and in vivo model of perinatal HI brain injury, Conclusion : These results may suggest that the administration of MPA before HI insult could significantly protect against perinatal HI brain injury via anti-apoptotic mechanisms, which offers the possibility of MPA application for the treatment of neonatal HI encephalopathy.

• Tuberculosis and Respiratory Diseases
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• v.47 no.6
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• pp.797-806
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• 1999

Background: About 20% of small cell lung cancer(SCLC) patients have bone marrow(EM) metastasis at the time of diagnosis and the remaining patients are also considered with micrometastasis. In an attempt to detect EM micrometastasis, we used cytokeratin(CK)-20 as a molecular marker, which is specific for epithelial cells. Method: A sensitive RT-PCR assay was used to compare CK-20 expression both in SCLC cell line H209 and normal leukocyte and to evaluate EM aspirates of 28 SCLC patients. Result: H209 cell line showed CK-20 expression but normal leukocyte did not, suggesting CK-20 expression is lung tissue-specific. Of 28 patients(11 limited disease, 17 extensive disease), only 2(1/11, 1/17) samples tested revealed positive signal for CK-20. Two patients with CK-20 expression had EM metastasis or multiple bone involvement during follow-up. Conclusion: Although circulating tumor cells were detected in EM of small portion of patients with bone metastasis, CK-20 doesn't seem to be a reliable marker for the detection of micrometastasis in SCLC. This study emphasizes that identification of more specific marker for micrometastsis is mandatory prior to clinical application.