• Title/Summary/Keyword: water primer

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Construction of Improved PCR Primer Set for the Detection of Human Enteric Adenovirus 41

  • Cho, Kyu-Bong
    • Biomedical Science Letters
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    • v.24 no.3
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    • pp.230-238
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    • 2018
  • Human enteric Adenovirus-41 (HuEAdV-41) causes gastroenteritis, which detected by the polymerase chain reaction (PCR) base diagnostic system for clinical, food, environmental, fish and shellfish samples. We developed improved PCR and nested PCR primer set which had high specificity, sensitivity and reduced times. In this study, we compared seventeen conditions reported in the previous study that was using the PCR based HuEAdV-41 detection system, and non-enteric Adenovirus were detected in nine conditions. The most sensitive detection condition was up to 25 copies however it took 184 minutes of PCR reaction time. In this study, the PCR primer set developed had same level of sensitivity, it reduced the time of detection for clinical, food and seafood samples to 112 minutes. Developed nested PCR primer set needed 112 minutes but detected up to approximately 1 copy. In addition, developed PCR and nested PCR primer set was validated with twenty samples of underground water at random, of which ten samples showed specific band without non-specific reaction. We expect this study will be used to diagnose HuEAdV-41 from various samples.

An Experimental Evaluation on Performance of Surface Protector for Concrete Structures (콘크리트 구조물 표면 보호재의 성능에 대한 실험적 평가)

  • Nam, Yong-Hyuk;Chung, Young-Jun;Jang, Suk-Hwan;An, Young-Ki
    • Journal of the Korea institute for structural maintenance and inspection
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    • v.10 no.5
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    • pp.157-163
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    • 2006
  • This study is on the evaluation of double surface protection method using water repellent primer and final top coat to protect concrete. Water repellent agent has been applied on the final top coat to protect concrete. However, to make up for the weakness to the ultraviolet of the water repellent, the work procedure of these protectors is done vice versa. This combination of protectors was compared with existing ones in this study. Even though the final top coat was applied on the water repellent primer, its adhesive strength met to KS F 4936-' 03 with other protectors used in this study. All surface protectors used in this study were excellent in protecting concrete. Especially, in case of applying with final top coat in conjunction with water repellent primer, the resistance against chloride ion penetration and neutralization by $CO_2$ was more efficient than other surface protectors used in this study under this given condition.

Molecular Biological Detection of the Genes Encoding Aminoglycosise Acetyltransferases and Aerolysin in Water Samples from Juam Lake (주암호에서 Aminoglycoside Acetyltransferases와 Aerolysin 유전자의 분자생물학적 검출)

  • 이영종;한효심;정재성
    • Korean Journal of Microbiology
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    • v.36 no.4
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    • pp.273-278
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    • 2000
  • The aacCl, aacC2, aacC3, and aacC4 genes, which encode aminoglycoside acetyltransferase AAC(3)-I, AA(3)-II, AAC(3)-III, and AAC(3)-IV, respectively, and aerolysin genes in water samples from Juam lake were surveyed by polymerase chain reaction. Surface water was collected from January 1996 to December 1998, and then bacterial DNA was extracted from the water. Twelve samples were tested by PCR to servey the genes for aminoglycoside acetyltransferase and aerolysin in the lake water. The aacC2 gene was detected in 9 of 12 DNA samples. Among 9 samples showing aacC2 positive, 7 samples were associated with Tn3 sequence. However, none of the twelve samples amplified the expected DNA fragment for aacC1, aacC3, and aacC4 genes. PCR primer to detect the aerolysin gene was designed using the conserved region of the genes for aerolysin and hemolysin of Aeromonas spp. This primer set successfully amplified the expected 414 bp PCR product with the DNA samples from the lake water. The aerolysin gene was detected in 7 of 12 DNA samples. When Southern hybridization of the gel with probe was performed, the aerolysin gene was detected in 10 of 12 DNA samples. However, the seasonal fluctuation of these genes was not found.

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Quantitative Analysis of Human- and Cow-Specific 16S rRNA Gene Markers for Assessment of Fecal Pollution in River Waters by Real-Time PCR

  • Jeong, Ju-Yong;Park, Hee-Deung;Lee, Kyong-Hee;Hwang, Jae-Hong;Ka, Jong-Ok
    • Journal of Microbiology and Biotechnology
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    • v.20 no.2
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    • pp.245-253
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    • 2010
  • The base sequences representing human- and cow-specific 168 rRNA gene markers identified in a T-RFLP analysis were recovered from clone libraries. The human- and cow-specific primers were designed from these sequences and their specificities were analyzed with fecal DNAs from human, cow, and pig. The AllBac primer set showed positive results for all human, cow, and pig samples, whereas the human-specific primer set showed positive result only for the human sample but not for the cow or pig samples. Likewise, the cow-specific primer set showed positive results only for the cow sample but not for the human or pig samples. Real-time PCR assay with these primers was developed for the identification and quantification of fecal pollution in the river water. The human- and cow-specific markers were detected in the order of 9 $\log_{10}$ copies per gram wet feces, which were two orders of magnitude lower than those of total Bacteroidales. For the river water samples, the human-specific marker was detected in $1.7-6.2\;\log_{10}$ copies/100 ml water, which was 2.4-4.9 orders of magnitude lower than those of total Bacteroidales. There was no significant correlation between total Bacteroidales and conventional fecal indicators, but there was a high correlation between Bacteroidales and the human-specific marker. This assay could reliably identify and quantify the fecal pollution sources, enabling effective measures in the watersheds and facilitating water quality management.

Shear bond strength of Universal bonding systems to Ni-Cr alloy (니켈-크롬 합금에 대한 다용도 접착 시스템의 전단결합강도)

  • Song, So-Yeon;Son, Byung-Wha;Kim, Jong-Yeob;Shin, Sang-Wan;Lee, Jeong-Yol
    • The Journal of Korean Academy of Prosthodontics
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    • v.53 no.4
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    • pp.295-300
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    • 2015
  • Purpose: The aim of this study was to evaluate the shear bond strength between Ni-Cr alloy and composite resin using universal adhesive systems coMPared to conventional method using metal primers. Materials and methods: For this study, a total of 120 cast commercial Ni-Cr alloy (Vera Bond 2V) disks were embedded in acrylic resin, and their surfaces were smoothed with silicon carbide papers and airborne-particle abrasion. Specimens of each metal were divided into 6 groups based on the combination of metal primers (Metal primer II, Alloy primer, Metal & Zirconia primer, MKZ primer) and universal adhesive systems (Single Bond Universal, All Bond Universal). All specimens were stored in distilled water at $37^{\circ}C$ for 24 hours. Shear bond strength testing was performed with a universal testing machine at a cross head speed of 1 m/min. Data (MPa) were analyzed using one-way ANOVA and the post hoc Tukey's multiple comparison test (${\alpha}$=.05). Results: There were significant differences between Single Bond Universal, All Bond Universal, Metal Primer II and Alloy Primer, MKZ Primer, Metal & Zirconia Primer (P<.001). Conclusion: Universal Adhesive system groups indicated high shear bond strength value bonded to Ni-Cr alloy than that of conventional system groups using primers except Metal Primer II. Within the limitations of this study, improvement of universal adhesive systems which can be applied to all types of restorations is recommended especially non-precious metal alloy. More research is needed to evaluate the effect of silane inclusion or exclusion in universal adhesive systems.

A STUDY ON SHEAR BOND STRENGTH OF COMPOSITE RESIN TO DENTIN FOLLOWING SURFACE TREATMENTS (표면처리에 따른 상아질과 콤포짓드 레진간의 전단결합강도에 관한 연구)

  • Noh, Eun-Hee;Park, Sang-Jin
    • Restorative Dentistry and Endodontics
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    • v.16 no.1
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    • pp.200-208
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    • 1991
  • The purpose of this study was to observe shear bond strength of composite resin to dentin following surface treatment. Freshly extracted forty-eight sound human molars were used in this study. They were stored at $4^{\circ}C$ physiologic saline solution before experiment. The teeth was then mounted with self curing acrylic resin in brass mold. The buccal surfaces of the teeth were grinding approximately 1.5mm by means of water-irrigated grinding wheel to expose the flattened fresh dentin surfaces. The specimens were divided into 6 groups according to preparation and treatment procedures on dentin surfaces; Group 1: Untreated after preparation with No.301 diamond point Group 2: Treated with primer for 60 seconds after preparation with No.301 diamond point Group 3: Untreated after preparation with No.700 fissure carbide bur Group 4: Treated with primer for 60 seconds after preparation with No.700 fissure carbide bur Group 5: Untreated after grinding with 600 grit silicon carbide paper Group 6: Treated with primer for 60 seconds after grinding with 600 grit silicon carbide paper Light cure dental adhesive was applicated to each specimen. Silux plus(3M) was inserted then into polyethylene tube of 3mm diameter and 3mm height, and polymerized to dentin surface. All of the specimens were stored in distilled water at $35.6^{\circ}C$ for 24 hours prior to testing. The shear bond strength was measured using an Instron Universal Testing Machine. The results obtained from this study were as follows: 1. The shear bond strength to dentin was the highest in group II. 2. The shear bond strength to dentin was the lowest in group III. 3. There was no significant difference in shear bond strength to dentin according to preparation instrument. 4. The primer treatment group showed significantly greater shear bond strength than untreated group.

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THE EFFECT OF ADDITIONAL ENAMEL ETCHING ON MICROLEAKAGE OF THE ADHESION OF SELF-ETCHING PRIMER SYSTEM (자가 산부식 프라이머 시스템 사용시 인산에 의한 부가적인 산부식이 미세누출에 미치는 영향)

  • Yoon, Jung-Jin;Min, Kyung-San;Hong, Chan-Ui
    • Restorative Dentistry and Endodontics
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    • v.28 no.5
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    • pp.363-368
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    • 2003
  • The purpose of this study is to evaluate the effect of additional enamel etching with phosphoric acid on the microleakage of the adhesion of self-etching primer system. Class V cavity($4mm{\times}3mm{\times}1.5mm$) preparations with all margins in enamel were prepared on buccal surface of 42 extracted human upper central incisor teeth. Prepared teeth were randomly divided into 3 groups. Group 1:no additional pretreatment with 37% phosphoric acid (NE). Group 2:additional pretreatment with 37% phosphoric acid for 10 seconds (E10s). Group 3:additional pretreatment with 37% phosphoric acid for 20 seconds (E20s). The adhesives(Clearfil SE $Bond^{\circledR}$, Kuraray, Osaka, Japan) and composite resins(Clearfil $AP-X^{\circledR}$, Osaka, Kuraray, Japan) were applied following the manufacturer's instructions. All the specimens were finished with the polishing disc(3M dental product, St Paul, MN, USA), thermocycled for 500 cycles between $5^{\circ}C$ and $55^{\circ}C$ and resected apical 3-mm root. 0.028 stainless steel wire was inserted apically into the pulp chamber of each tooth and sealed into position with sticky wax. Surrounding tooth surface was covered with a nail varnish 2 times except areas 1mm far from all the margins. After drying for one day, soaked the samples in the distilled water. Microleakage was assessed by electrochemical method(System 6514, $Electrometer^{\circledR}$), Keithley, USA) in the distilled water. In this study, the microleakage was the lowest in group 1 (NE) and the highest in group 3(E20s)(NE

Efficiency Comparison of Environmental DNA Metabarcoding of Freshwater Fishes according to Filters, Extraction Kits, Primer Sets and PCR Methods (분석조건별 담수어류의 환경 DNA 메타바코딩 효율 비교: 필터, 추출 키트, 프라이머 조합 및 PCR 방법)

  • Kim, Keun-Sik;Kim, Keun-Yong;Yoon, Ju-Duk
    • Korean Journal of Ecology and Environment
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    • v.54 no.3
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    • pp.199-208
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    • 2021
  • Environmental DNA (eDNA) metabarcoding is effective method with high detection sensitivity for evaluating fish biodiversity and detecting endangered fish from natural water samples. We compared the richness of operational taxonomic units(OTUs) and composition of freshwater fishes according to filters(cellulose nitrate filter vs. glass fiber filter), extraction kits(DNeasy2® Blood & Tissue Kit vs. DNeasy2® PowerWater Kit), primer sets (12S rDNA vs. 16S rDNA), and PCR methods (conventional PCR vs. touchdown PCR) to determine the optimal conditions for metabarcoding analysis of Korean freshwater fish. The glass fiber filter and DNeasy2® Blood & Tissue Kit combination showed the highest number of freshwater fish OTUs in both 12S and 16S rDNA. Among the four types, the primer sets only showed statistically significant difference in the average number of OTUs in class Actinopterygii (non-parametric Wilcoxon signed ranks test, p=0.005). However, there was no difference in the average number of OTUs in freshwater fish. The species composition also showed significant difference according to primer sets (PERMANOVA, Pseudo-F=6.9489, p=0.006), but no differences were observed in the other three types. The non-metric multidimensional scaling (NMDS) results revealed that species composition clustered together according to primer sets based on similarity of 65%; 16S rDNA primer set was mainly attributed to endangered species such as Microphysogobio koreensis and Pseudogobio brevicorpus. In contrast, the 12S rDNA primer set was mainly attributed to common species such as Zacco platypus and Coreoperca herzi. This study provides essential information on species diversity analysis using metabarcoding for environmental water samples obtained from rivers in Korea.

Evaluation of Adhesive Performance of Surface Finishing Material with Primer Based on Silane (실란계 프라이머를 활용한 바닥 마감재 부착성능 평가)

  • Jeong, Gwon-Young;Youn, Da Ae;Jang, Seok-Joon;Kil, Bae-Su;Yun, Hyun-Do
    • Journal of the Korea institute for structural maintenance and inspection
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    • v.21 no.4
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    • pp.39-46
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    • 2017
  • The experimental research was conducted to evaluate the adhesive performance of surface finishing material with primer based on silane(primer). For this purpose, concrete specimens with compressive strength of 18, 30, 50 MPa were made and cured in water condition ($20{\pm}2^{\circ}C$) for 28 days. A primer was applied on the age of 28 days and evaluated according to based on the curing age of the surface finishing material. Moreover, the mortar specimen also made and tested as per KS F 4937 for compared with concrete-based test results. Test results indicated that the adhesive strength of specimens with primer exhibit similar than that of specimens without primer. Also, the adhesive performance improved with increasing in curing age and compressive strength. The correlation between compressive and adhesive strength of mortar and concrete specimens showed similar trend. It was noted that there is no significant effects of primer on adhesive performance of surface finishing material, thus use of primer has superior potential for solving durability problem of concrete slab surface.