Kim, Tae-Woon;Jang, Yun-Ho;Jeong, Min Kyu;Seo, Yoonjeong;Park, Chan Ho;Kang, Sinseok;Lee, Young Ju;Choi, Jeong-Soo;Yoon, Soon-Seek;Kim, Jae Myung
Journal of Veterinary Science
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v.22
no.2
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pp.24.1-24.16
/
2021
Background: Bovine tuberculosis (TB) is caused by Mycobacterium bovis, a well-known cause of zoonotic tuberculosis in cattle and deer, and has been investigated in many physiological and molecular studies. However, detailed genome-level studies of M. bovis have not been performed in Korea. Objectives: To survey whole genome-wide single-nucleotide polymorphism (SNP) variants in Korean M. bovis field isolates and to define M. bovis groups in Korea by comparing SNP typing with spoligotyping and variable number tandem repeat typing. Methods: A total of 46 M. bovis field isolates, isolated from laryngopharyngeal lymph nodes and lungs of Korean cattle, wild boar, and Korean water deer, were used to identify SNPs by performing whole-genome sequencing. SNP sites were confirmed via polymerase chain reaction using 87 primer pairs. Results: We identified 34 SNP sites with different frequencies across M. bovis isolates, and performed SNP typing and epidemiological analysis, which divided the 46 field isolates into 16 subtypes. Conclusions: Through SNP analysis, detailed differences in samples with identical spoligotypes could be detected. SNP analysis is, therefore, a useful epidemiological tracing tool that could enable better management of bovine TB, thus preventing further outbreaks and reducing the impact of this disease.
Artemisinin resistance (ART) has been confirmed in Greater Mekong Sub-region countries. Currently, C580Y mutation on Pfkelch13 gene is known as the molecular marker for the detection of ART. Rapid and accurate detection of ART in field study is essential to guide malaria containment and elimination interventions. A simple method for collection of malaria-infected blood is to spot the blood on filter paper and is fast and easy for transportation and storage in the field study. This study aims to evaluate LAMP-SNP assay for C580Y mutation detection by introducing an extra mismatched nucleotide at the 3' end of the FIP primer. The LAMP-SNP assay was performed in a water bath held at a temperature of 56℃ for 45 min. LAMP-SNP products were interpreted by both gel-electrophoresis and HNB-visualized changes in color. The method was then tested with 120 P. falciparum DNA from dried blood spot samples. In comparing the LAMP-SNP assay results with those from DNA sequencing of the clinical samples, the 2 results fully agreed to detect C580Y. The sensitivity and specificity of the LAMP-SNP assay showed 100%. There were no cross-reactions with other Plasmodium species and other Pfkelch13 mutations. The LAMP-SNP assay performed in this study was rapid, reliable, and useful in detecting artemisinin resistance in the field study.
This study evaluated the effects of various cleaning methods on the shear bond strength of zirconia ceramics after saliva contamination. Eighty zirconia disk specimens were divided into 8 groups. All groups were treated with one coat of MDP primer. All specimens (except the negative control) were contaminated with human saliva on the zirconia surface. The positive control went through the bonding procedure immediately after contamination without any cleaning procedure. With the exception of control groups, the remaining six groups were rinsed with water and either applied with MDP recoating (WATER+MDP) or without MDP recoating (WATER). While some were cleaned with a Ivoclean with MDP recoating (IVOCLEAN+MDP) or not applied with MDP recoating(IVOCLEAN), others were cleaned with a 1% NaOCl solution with MDP recoating (NaOCl+MDP) or without MDP recoating (NaOCl). The shear bond strength of all specimens were measured after being stored in distilled water at $37^{\circ}C$ for 24 hours. The data was analyzed statistically by an analysis of ANOVA, Tukey's post hoc test and Student's t-test was used to compare the shear bond strength according to the re-coating of MDP after the cleaning procedure. The positive control group showed the lowest shear bond strength value, and the WATER group and NaOCl group showed no significant difference when compared to the positive control group. The IVOCLEAN group showed significantly higher shear bond strength when compared to Water group and NaOCl group but not with the group of negative control. After rinsing with water or the NaOCl solution, the comparison of the single coating of MDP and re-coating of MDP showed different shear bond strengths but there was no significant difference to the negative control. After rinsing with Ivoclean, there was no significant difference to the negative control regardless of the recoating of MDP. In conclusion, the shear bond strength was affected by the cleansing procedure and Ivoclean was found to be effective regardless of the re-coating of MDP. When water or the NaOCl solution is used to remove surface contaminants, the re-coating of MDP provides a positive effect on cementation.
The purpose of this study was to investigate the effect of phosphoric acid concentration on the movement of 2-hydroxyethylmethacrylate(HEMA) from bonding resin - resin composite combination through dentin in vitro. Freshly extracted human third molar teeth were divided into four groups each of 10 teeth. A closed chamber with 1 ml distilled water was attached to the CEJ of each tooth. An occlusal cavity of 4mm diameter & remaining dentin thickness of 1.0-1.5mm was prepared in each tooth. Dentin was treated with 10% phosphoric acid gel for 15 seconds. 32% phosphoric acid gel for 15 seconds, or with 35% phosphoric acid gel for 15 seconds. A control group not treated with acid gel was also prepared. The cavities were rinsed, dried and then treated with the HEMA-containing All-Bond 2 primer & bonding resin which was light-cured for 10 seconds. The cavities were then restored with Z100 composite resin(shade:A3.5:3M Dent. Prod. USA) & light cured for 30 seconds. Water samples were retrieved from the chambers over a time course (4.32, 14.4, 43.2, 144 & 432 minutes ; 1, 3 & 10 days) and analyzed by high performance liquid chromatography. The results were as follows. 1. HEMA was detected in the pulp chambers of all teeth from 4.32 minutes after resin placement The highest rate of release was in the first sample period (0-4.32 min) & rate of release declined exponentially thereafter. 2. No significant differences were found for mean release rate for HEMA over a time course among the four groups (p>0.05). 3. The diffusion rate was significantly (p<0.05) less for 10% phosphoric acid gel than 32% phosphoric acid gel at the second sample period(4.32-14.4 min). 4. No significant differences were found for cumulative HEMA diffusion among the four groups at 10 days(p>0.05) and mean total(cumulative) release at 10 days for all groups was in the 9 - 16 nmol range. 5. The cumulative release was significantly (p<0.05) less for 10% phosphoric acid gel than 32% phophoric acid gel at the third(14.4-43.2 min) & fourth(43.2-144 min) sample period.
Journal of the Korean Society of Environmental Restoration Technology
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v.23
no.2
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pp.69-89
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2020
Scientific trust and quantification of traditional species investigation and results that have been used in ecology for decades has always been a problem and concern for ecologists. Global ecologists have proposed DNA-based species investigation studies to find answers to problems. In this study, we reviewed the global trend of research on environmental DNA(eDNA), which is a method for monitoring species by detecting DNA of organisms naturally mixed in environmental samples such as water, soil, and feces. The first eDNA research confirmed the possibility of species investigation at the molecular level, and commercialization of NGS(Next Generation Sequencing) and DNA metabarcoding elicits efficient and quantitative species investigation results, and eDNA research is increasing in the filed of ecology. In this study, mammals and birds were detected using MiMammal universal primers from 23 samples(3 natural reserves; 20 water bowls) out of 4 patches to verify eDNA for urban ecosystems in Suwon, and eDNA was verified by performing camera trapping and field survey. Most terrestrial species were detected through eDNA, and particularly, mice(Mus musculus), and Vinous-throated Parrotbill (Sinosuthora webbiana) were identified only with eDNA, It has been confirmed to be highly effective by investigating techniques for small and internal species. However, due to the lack of resolution of the primer, weasels(Mustela sibirica) and squirrels(Melanochromis auratus) were not detected, and it was confirmed that the traditional investigation method was effective only for a few species, such as Mogera robusta(Mogera robusta). Therefore, it is judged that the effects of species investigation can be maximized only when eDNA is combined with traditional field survey and Camera trapping to complement each other.
The composites(PM-A-PM-I) of 9 groups containing 7 kinds of hot water extracted medicinal plants were produced and evaluated its antioxidative activity. Each medicinal plants used these composites were analyzed in primer research that its anti oxidative activity was high. In the composites of medicinal plant, electron donating ability was higher than 70% in all sample. PM-D, PM-E and PM-F were significantly higher than others. Reducing power have similar tendency to electron donating ability. PM-D was the strongest in hydroxyl radical scavenging activity, followed by PM-E. In linoleic acid system, antioxidative activity of all sample was showed more than 50%, except PM-A. Especially PM-E and PM-F have significantly higher activity. Nitrite scavenging effect was significantly increased by PM-D, PM-E and PM-F added to Inula Japonica Thunberg. In these results, we suggested that high-er antioxidative activity of PM-D, PM-E and PM-F may be responsible for the contents of phenolic compounds present in Inula Japonica Thunberg. And thought to be enhanced by the synergy effect of the water extracted medicinal plants in the composite.
In this study, adaptation of compomer to saliva contaminated dentin was evaluated with scanning electron microscope(SEM) and confocal laser scanning microscope(CLSM). For the SEM study, the occulusal surfaces of thirty two molar teeth were grounded to exposure dentin surfaces. The specimen were randomly assigned to control and three experimental groups with four samples in each group. In control group, Dyract and F-2000 compomer were bonded on the specimens according to the manufactures direction. Experimental groups were subdivided into three groups. They were contaminated with saliva on dentin surfaces ; Experimental group 1 : Saliva was dried with compressed air. Experimental group 2 : Saliva was rinsed with air-water spray and dried. Experimental group 3 : After polymerization of an adhesive, they were contaminated with saliva, and then saliva was rinsed with air-water spray and dried. Dyract and F-2000 compomer were bonded on saliva-treated dentin surfaces. The interfaces between dentin and compomer were observed with SEM. For the CLSM study, Class V cavities were prepared in buccal and ligual surfacess of thirty two molars. The specimens were divided into control and experimental groups. Class V cavities in experimental group were contaminated with saliva and those surfaces in each experimental groups received the same treatments as for the SEM study. Cavities were applied Prime & Bond 2.1 and F-2000 compomer primer/adhesive that were mixed with fluorescein, and then were filled with Dyract and F-2000 compomer. Specimens were embedded in transparent acrylic resin and sectioned buccolingual1y with diamond wheel saw, and then mounted on cover slide for CLSM study. The interface between cavity and compomer was observed by fluoresence imaging with a CLSM. The results were as follows : 1. In SEM exammination of Dyract group, control group, experimental group 2, 3 showed close adaptation to dentin and hybrid layer of $3{\sim}4{\mu}m$ diameter. Interfacial gap between compomer and dentin in experimental group 1 was wider than in control group. 2. In SEM examination of F-2000 group, adaptation to dentin of control group was closer than Dytact control group, but hybrid-like layer was not observed. Interfacial gap between compomer and dentin in experimental group 1 was wider than in Dyract experimental group 1. 3. In dissolution specimens of Dyract and F-2000 group, resin tags penetrated through dentinal tubules in control group and experimental group 1 and 3, but the penetration of resin tag was irregular and partial in experimental group 1. 4. In CLSM exammination of Dyract and F-2000 group, adhesive patterns of control and experimental groups showed same as in SEM. This result suggests the treatment methods, rinsing & drying, repeating all adhesive procedures, will produce good effect on adaptation of compomer to dentin if the dentin surface or polymerized adhesive is contaminated by saliva.
Kim, Baek Jun;Lee, Yun-Sun;An, Jung-hwa;Park, Han-Chan;Okumura, Hideo;Lee, Hang;Min, Mi-Sook
Molecules and Cells
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v.26
no.3
/
pp.314-318
/
2008
Korean long-tailed goral (Nemorhaedus caudatus) is one of the most endangered species in South Korea. However, detailed species distribution and sex ratio data on the elusive goral are still lacking due to difficulty of identification of the species and sex in the field. The primary aim of this study was to develop an economical PCR-RFLP method to identify species using invasive or non-invasive samples from five Korean ungulates: goral (N. caudatus), roe deer (Capreolus pygargus), feral goat (Capra hircus), water deer (Hydropotes inermis) and musk deer (Moschus moschiferus). The secondary aim was to find more efficient molecular sexing techniques that may be applied to invasive or non-invasive samples of ungulate species. We successfully utilized PCR-RFLP of partial mitochondrial cytochrome b gene (376 bp) for species identification, and sex-specific amplification of ZFX/Y and AMELX/Y genes for sexing. Three species (goral, goat and water deer) showed distinctive band patterns by using three restriction enzymes (Xbal, Stul or Sspl). Three different sexing primer sets (LGL331/335 for ZFX/Y gene; SE47/48 or SE47/53 for AMELX/Y gene) produced sex-specific band patterns in goral, goat and roe deer. Our results suggest that the molecular analyses of non-invasive samples might provide us with potential tools for the further genetic and ecological study of Korean goral and related species.
Journal of The Korean Society of Grassland and Forage Science
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v.34
no.2
/
pp.114-119
/
2014
Salinity and drought stresses are probably the most significant abiotic factor limiting plant's growth, also negatively affect seed germination and early seedling development. To study on effect of NaCl and PEG stress on seed germination and gene expression pattern of tall fescue, the levels of NaCl and PEG-induced water stresses were determined in first experiment. Different concentration of NaCl (0 to 350 mM) and PEG (0 to 30%) were used for seed treatment. Seed Germination percentage reduced with increasing osmotic potential of growth medium either due to NaCl or PEG. Seeds were not germinate at 350 mM NaCl or 30% PEG treatment. On the basis of the results, Kentucky31(E-) had more resistant than Fawn in both stress conditions. Furthermore, we have used an annealing control primer-based differential display reverse transcription-polymerase chain reaction method to identify salt- and drought stress-induced differentially expressed genes (DEGs) in tall fescue leaves. Using 120 annealing control primers, a total of 4 genes were identified and sequenced. The possible roles of the identified DEGs are discussed in the context of their putative role during salinity and drought stresses.
This study investigated the influence of IRM on marginal microleakage of 5th generation adhesives. Class V cavities with gingival margins in dentin were prepared on both buccal and lingual surfaces of 60 extract-ed human molar teeth. Prepared teeth were randomly divided into six groups. Group 1 and 4 received no temporary restoration with IRM. Group 2 and 5 were covered with IRM mixed at P/L ratio(10g/1g). Group 3 and 6 were covered with IRM mixed at P/L ratio(10g/2g). The temporary restorations were removed mechanically with an ultrasonic scaler after one-week storage in distilled water. The cavities were restored using one of two adhesives and composites ; Single Bond/Filtek Z 250(Croup 1, 2 and 3), UniFil Bond/UniFil F(Group 4, 5 and 6). Following one day storage in distilled water, the restored teeth were thermocycled for 500 cycles(between $5^{\circ}C{\;}and{\;}55^{\circ}C$) and immersed in 2 % methylene blue for dye penetration testing. The results were analysed using Kruskal-Wallis Test, Mann-Whitney and Wilcoxon signed ranked test at a significance level of 0.05. The results of this study were as follows 1. Ranking of mean microleakage scores at the enamel margins was Group 10.05) 4. At the dentin margins, the microleakage of the group not pretreated with IRM was lower than that of the group pretreated with IRM. And the microleakage of UniFil Bond was lower than that of Single Bond. 5. Compared with microleakages between the enamel and dentin margins of each groups, Group 1, 2, 3, 4, 5 and 6 at dentin margin were higher microleakage than those at enamel margin. There were significant difference between enamel and dentin microleakage of Group 2 and 3(p<0.05).
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