• 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.

• Korean Journal of Microbiology
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• v.41 no.3
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• pp.216-224
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• 2005

Chromatography has now been used successfully to provide the requisite purity for human plasma-derived biop-harmaceuticals such as coagulation factors and immunoglobulins. Recently, increasing attention has been focused on establishing efficient cleaning procedures to prevent potential contamination by microorganisms as well as carry-over contamination from batch to batch. The purpose of present study was to develop a cleaning validation system for the assurance of virus removal and/or inactivation during chromatography process. In order to establish an assay system for the validation of virus clearance during chromatography cleaning process, a quantitative real-time PCR method for porcine parvovirus(PPV) was developed, since PPV, a model virus for human parvovirus B19, has a high resistance to a range of physico-chemical treatment. Specific primers for amplification of PPV DNA was selected, and PPV DNA was quantified by use of SYBR Green I. The sensitivity of the assay was calculated to be 1.5 $TCID_{50}/ml$. The established real-time PCR assay was successfully applied to the validation of PPV removal and cleaning during SP-Sepharose cation chromatography for thrombin purification and Q-Sepharose anion chromatography for factor VIII purification. The comparative results obtained by real-time PCR assay and infectivity titrations suggested that the real-time PCR assay could be a useful method for chromatography cleaning validation and that it could have an additive effect on the interpretation and evaluation of virus clearance during the virus removal process.

• Journal of Food Hygiene and Safety
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• v.27 no.2
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• pp.146-151
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• 2012

In this study, effective gene extraction methods were compared to identify raw materials of processed meat products through molecular biological methods. Species specific primers were used to identify ingredients of processed foods and, as a sample, 13 kinds of processed meat products including beef, pork and chicken. According to the type of sample, 13 kinds of samples were classified into liquid type, source type and powder type. The samples were pre-treated (centrifugation) and (or) performed Whole Gene Amplification (WGA) kit for amplification of the extracted DNA. As a result, it was possible to identify the raw material of products through the centrifugation of sample 1 ml for liquid type of processed meat products. For source type of products after gene extraction, it was required to perform WGA for the identification of ingredients. For powder type products did not required any further pre-treatment and WGA. In this study, it was an opportunity to confirm the possibility of identification of raw material from the gene extraction of processed meat products and this method could be used to examine the authenticity of raw material of products.

• 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.

• Microbiology and Biotechnology Letters
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• v.33 no.2
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• pp.96-105
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• 2005

For screening of autoregulator receptor gene from Streptomyces longwoodensis, PCR was performed with primers of receptor gene designed on the basis of amino acid sequences of autoregulator receptor proteins with known function. PCR products were subcloned into the BamHIsite of pUC19 and transformed into the E. coli $DH5{\alpha}$. The isolated plasmid from transformant contained the fragment of 100 bp, which was detected on $2\%$ gel after BamHI treatment. The insert, 100 bp PCR product, was confirmed as the expected internal segment of gene encoding autoregulator receptor protein by sequencing. Southern and colony hybridizations with the 100 bp fragment as a probe allowed to select a genomic clone of S. longwoodensis, pSLT harboring a 4.4 kb SphI fragment. Nucleotide sequencing analyses revealed a 651 bp open reading frame(ORF) were isolated protein showing moderate homology ($35{\sim}46\%$) with the ${\Gamma}$-butyrolactone autoregulator receptors from Streptomyces sp., and this ORF was named sltR The sltR/pET-17b plasmid was constructed to overexpress the recombinant SltR protein (rSltR) in E. coli BL21 (DE3)/pLysS, and the rSltR protein was purified to homogeneity by DEAE-Sephacel column chromatography, and DEAE-5PW chromatography (HPLC). The molecular mass of the purified rSltR protein was 55 kDa by HPLC gel-filtration chromatography and 28 kDa by SDS-PAGE, indicating that the rSltR protein is present as a dimer. A binding assay with tritium-labeled autoregulators revealed that the rSltR has clear binding activity with a A-factor type autoregulator as the most effective ligand.

• Tuberculosis and Respiratory Diseases
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• v.43 no.6
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• pp.842-851
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• 1996

Background : Rifampicin(RFP) is a key component of the antituberculous shon-course chemotherapy and the RFP-resistance is a marker of multi-drug resistant(MDR) M. tuberculosis. rpoB gene encodes the ${\beta}$-subunit of RNA polymerase of M. tuberculosis which is the target of RFP. Recent reports show that rpoB gene mutations are the cause of RFP resistance of M. tuberculosis and the main mechanism of rpoB gene mutation is point mutation. And PCR-SSCP is a rapid and easy method for detecting point mutations. So we performed PCR-SSCP of rpoB gene of M. tuberculosis and compared the result with traditional RFP sensitivity test. Method : The 27 RFP sensitive M. tuberculosis culture isolates and 25 RFP resistant isolates were evaluated. The RFP sensitivity test was done at the Korean Tuberculosis istitute. The DNA was extracted by bead beater method and was amplified with primers TR-8 and TR-9 in a 20ul PCR reaction containing 0.1ul(luCi) [${\alpha}-^{32}P$] - dCTP. After amplification, SSCP was done using non-denaturaring polyacrylamide gel electrophoresis. Then direct sequencing was done in cases of different eletrophoretic mobility compared with that of H37Rv. In 19 cases, we compared PCR-SSCP results with patient's clinical course and the results of traditional RFP sensitivity test. Results : 1) All 27 RFP sensitive M. tuberculosis isolates showed the same electrophoretic mobility compared with that of H37Rv. And all 25 RFP resistant M. tuberculosis isolates showed different electrophoretic mobility. 2) The mechanism of rpoB gene mutation of M. tuberculosis is mainly point mutation. 3) The PCR-SSCP results correlate well with traditional RFP sensitivity and patient's clinical response to antituberculous treatment. Conclusion: The PCR-SSCP of rpoB gene is a very sensitive and rapid mehod in detecting RFP- resistant M. tuberculosis.

• Restorative Dentistry and Endodontics
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• v.18 no.1
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• pp.73-83
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• 1993

The purpose of this study was to evaluate the effect of various acid treatments on dentin bonding. Freshly extracted human teeth were uprightly embedded in self curing acrylic resin, and their occlusal surfaces were grinded to expose flat dentin surfaces. The specimens were divided into 4 groups. Specimens of one group were not treated so as to be a control and those of the other three groups were threated with 10% polyacrylic acid, 10% phosphoric acid, and 10-3 solution(10% citric acid/3% ferric chloride) respectively. Primer, bonding resin and composite resin were applied over the treated dentin surfaces sequentially. All specimens were stored in $37^{\circ}C$ distilled water for 24 hours, then the tensile bond strength was measured and the treated dentin surfaces and fracured dentin surfaces were examined under a scanning electron microscope. The results were as follows: Bond strengths of acid-treated groups were higher than those of the untreated group. In the acid-treated groups, bond strength was found to be the highest in the 10-3 solution group followed by the 10% phosphoric acid group and the 10% polyacrylic acid group(P<0.01). On SEM examination of dentin surfaces, the untreated dentin surface showed a remaining smear layer and closed dentinal tubules. Dentin surfaces treated with 10 % polyacrylic acid showed a clean dentin surface without the smear layer, but showed remaining smear plugs in dentinal tubules. A dentin surface treated with 10% phosphoric acid or 10-3 solution showed open dentinal tubules without the smear layer or smear plugs. On SEM observation of the fractured dentin-resin interface, the untreated group showed that failure occurred in the smear layer. The group treated with 10% polyacrylic acid showed no resin tag remained in the dentinal tubules, but resin tags in the dentinal tubules were observed in the group treated with the 10% phosphoric acid or the 10-3 solution. On the failure mode examination, the higher the bond strength of the group, the higher the frequency of cohesive failure. The coefficient between bond strength and cohesive failure rate was 0.71.