• Journal of Veterinary Science
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• 제22권2호
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• pp.24.1-24.16
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• 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.

• Parasites, Hosts and Diseases
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• 제59권1호
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• pp.15-22
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• 2021

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.

• 대한치과재료학회지
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• 제44권1호
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• pp.61-68
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• 2017

본 연구는 MDP 적용 후 타액으로 오염된 지르코니아 수복물을 다양한 방법으로 세척한 후 전단결합강도를 비교하여 세척방법이 결합강도에 미치는 영향을 알아보고자 한다. 80개의 지르코니아 시편을 8개의 군으로 나누었다. 모든 시편에 MDP를 적용한 후 한 개의 군(음성대조군)을 제외하고 나머지 군에 타액을 적용하여 오염시켰다. 그 중 한 개의 군(양성대조군)은 타액 오염 후 세척하지 않고 즉시 레진 시멘트를 이용하여 접착하였다. 나머지 6개의 군의 시편을 물을 이용하여 세척하고 MDP를 적용하거나(물+MDP) 적용하지 않은 군(물), Ivoclean으로 세척하고 MDP를 적용하거나(IVOCLEAN+MDP) 적용하지 않은 군(IVOCLEAN), 차아염소산나트륨을 이용하여 세척하고 MDP를 적용하거나(NaOCl+MDP) 적용하지 않은 군(NaOCl)으로 분류하였다. 모드 시편은 $37^{\circ}C$ 증류수에 24시간 저장한 후 전단강도를 측정하였고, ANOVA, Tukey's post hoc test를 이용하여 전단강도를 분석하였고, MDP의 재적용 여부가 미치는 영향에 대해서는 student t-test를 이용하여 통계분석하여 다음의 결과를 얻었다. 양성대조군이 가장 낮은 전단강도 값을 나타냈으며, 물군과 NaOCl군이 낮은 전단강도 값을 나타내며 양성대조군과 유의한 차이가 없었다. IVOCLEAN군은 물군과 NaOCl군보다 유의하게 높은 전단강도 값을 나타내며 음성대조군과 유의한 차이가 없었다. MDP를 재적용한 것은 물과 차아염소산나트륨을 이용한 경우 MDP를 재적용하지 않은 경우와 유의한 차이를 나타내면서 음성대조군과 유의한 차이가 없었다. Ivoclean을 사용한 경우 MDP 재적용 여부와는 관계없이 음성대조군과 유의한 차이가 없었다. 결론적으로, 세척방법에 따라 전단강도는 영향을 받으며, MDP 재적용 여부와는 관계없이 Ivoclean이 효과적이며, 물과 차아염소산나트륨 사용시에는 MDP를 다시 적용해주는 것이 결합강도를 향상시킬 수 있는 방법으로 사료된다.

• Restorative Dentistry and Endodontics
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• 제24권1호
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• pp.147-155
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• 1999

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.

• 한국환경복원기술학회지
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• 제23권2호
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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.

• 생명과학회지
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• 제17권8호통권88호
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• pp.1135-1140
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• 2007

항산화 활성이 높은 한약재 10종을 대상으로 9종의 조성물(PM-A-PM-I)을 제조하여 항산화 활성을 비교 분석하였다. 한약재 조성물의 DPPH에 대한 전자공여능은 모든 실험군에서 70%이상이었으며, 조성물 중 PM-D, PM-E 및 PM-F가 여타 시료에 비해 유의적으로 전자공여능이 높았다. 환원력도 전자공여능과 동일한 경향이었다. Hydroxyl radical에 대한 항산화 활성은 PM-D가 가장 높았고 다음으로 PM-E였 다. Linoleic acid 에 대한 자동산화 억제능은 PM-A를 제외한 모든 시료에서 50%이상의 활성을 나타내었으며, PM-E, PM-F에서 유의적으로 활성이 높았다. 아질산염 소거능은 선복화가 첨가된 PM-D, PM-E, PM-F에서 유의적으로 높았다. 따라서 9종의한약재 조성물의 항산화활성은 첨가된 한약재의 종류가 유사하였으나 PM-D, PM-E, PM-F 에서 높은 항산화활성을 보인 것은 선복화의 작용이 컸던 것으로 생각되며, 또한 한약재에 함유된 항산화 성분간의 시너지 효과 때문인 것으로 추정된다.

• Restorative Dentistry and Endodontics
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• 제25권4호
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• pp.575-586
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• 2000

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.

• Molecules and Cells
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• 제26권3호
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• pp.314-318
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• 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.

• 한국초지조사료학회지
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• 제34권2호
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• pp.114-119
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• 2014

염 또는 건조 스트레스 처리에 의한 톨 페스큐 종자의 발아율 변화와 유식물체 수준에서의 유전자 발현을 조사하기 위하여 in vitro 조건에서 NaCl과 PEG를 처리하여 분석하였다. NaCl 처리시 톨 페스큐 품종별 발아율은 50 mM 농도에서 발아율이 서서히 감소하기 시작하였으며 350 mM의 농도에서는 모든 품종에서 발아가 되지 않는 경향을 보였다. NaCl 처리 농도에 따른 발아율 감소율은 Fawn 품종이 가장 큰 변화를 보였으며 Kentucky-31(E-) 품종이 가장 강한 내성을 보였다. 또한, PEG 처리시 톨 페스큐 품종별 발아율의 변화도 NaCl 처리시와 유사한 경향을 보였으며 고농도인 30% PEG 처리구에서는 모든 품종에서 발아가 되지 않는 경향을 보였으며 Kentucky-31(E-) 품종이 가장 강한 내성을 보였다. 톨 페스큐 유식물체 수준에서 염해와 건조 스트레스에 의한 유전자 발현양상을 조사하기 위하여 DEGs (differentially expressed genes) 탐색을 위한 ACP-based GeneFishing$^{TM}$ PCR 분석을 통해 NaCl 또는 PEG 처리에 따른 발현량의 차이를 보이는 총 4개의 DEG를 선발하여 클로닝하고 염기서열을 분석하였다. 무처리구에 비해 NaCl 처리시 4개의 DEG가 증가하였고 감소하는 DEG는 확인 되지 않았으나, PEG 처리에서는 3개의 DEG (DEG 1, 3, 및 4)가 증가하였고 1개의 DEG가 감소하는 경향을 나타내었다. 발굴된 DEG들을 blastx 검색에 의하여 rubisco large subunit (DEG1), microsomal glutathion S-transferase (GST) 3-like isoform 1 (DEG2) 유전자로 동정되었다.

• Restorative Dentistry and Endodontics
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• 제28권1호
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• pp.1-10
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• 2003

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).