• Korean Journal of Veterinary Research
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• v.54 no.2
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• pp.113-115
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• 2014

Salmonella are causative agents of gastroenteritis and systemic disease in animals. The invA gene was selected as a target sequence of loop-mediated isothermal amplification (LAMP) assay for diagnosis of Salmonella infection. The detection limits for broth dilution, spiked feces and enrichment were $10^4$, $10^5$ and $10^2$ CFUs/mL, respectively. The LAMP assay developed in the present study may be a reliable method for detection of Salmonella spp. in pig feces.

• The Plant Pathology Journal
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• v.32 no.5
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• pp.414-422
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• 2016

Eucalyptus dieback disease, caused by Cylindrocladium scoparium, has occurred in last few years in large Eucalyptus planting areas in China and other countries. Rapid, simple, and reliable diagnostic techniques are desired for the early detection of Eucalyptus dieback of C. scoparium prior to formulation of efficient control plan. For this purpose, three PCR-based methods of nested PCR, multiplex PCR, loop-mediated isothermal amplification (LAMP) were developed for detection of C. scoparium based on factor 1-alpha (tef1) and beta-tubulin gene in this study. All of the three methods showed highly specific to C. scoparium. The sensitivities of the nested PCR and LAMP were much higher than the multiplex PCR. The sensitivity of multiplex PCR was also higher than regular PCR. C. scoparium could be detected within 60 min from infected Eucalyptus plants by LAMP, while at least 2 h was needed by the rest two methods. Using different Eucalyptus tissues as samples for C. scoparium detection, all of the three PCR-based methods showed much better detection results than regular PCR. Base on the results from this study, we concluded that any of the three PCR-based methods could be used as diagnostic technology for the development of efficient strategies of Eucalyptus dieback disease control. Particularly, LAMP was the most practical method in field application because of its one-step and rapid reaction, simple operation, single-tube utilization, and simple visualization of amplification products.

• Pediatric Infection and Vaccine
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• v.22 no.2
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• pp.106-112
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• 2015

Purpose: The long-term administration of antibiotics interferes with bacterial culture in the middle ear fluids (MEFs) of young children with otitis media with effusion (OME). The purpose of this study is to determine whether molecular diagnostics can be used for rapid and direct detection of the bacterial pathogen in culture-negative MEFs. Methods: The specificity and sensitivity of both polymerase chain reaction (PCR) and loop-mediated isothermal amplification (LAMP) to the lytA gene of Streptococcus pneumoniae were comparatively tested and then applied for pneumococcal detection in the clinical MEFs. Results: The detection limit of the PCR assay was approximately $10^4$ colony forming units (CFU), whereas that of LAMP was less than 10 CFU for the detection of S. pneumoniae. Both PCR and LAMP did not amplify nucleic acid at over $10^6$ CFU of H. influenzae or M. catarrhalis, both of which were irrelevant bacterial species. Of 22 culture-negative MEFs from children with OME, LAMP positivity was found in twelve MEFs (54.5%, 12/22), only three of which were PCR-positive (25%, 3/12). Our results showed that the ability of LAMP to detect pneumococcal DNA is over four times higher than that of PCR (P<0.01). Conclusions: As a high-resolution tool able to detect nucleic acid levels equivalent to <10 CFU of S. pneumoniae in MEFs without any cross-reaction with other pathogens, lytA -specific LAMP may be applied for diagnosing pneumococcus infection in OME as well as evaluating the impact of a pneumococcal conjugate vaccine against OME.

• International Journal of Industrial Entomology and Biomaterials
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• v.48 no.3
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• pp.139-146
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• 2024

For stable silkworm breeding and high-quality sericulture product production, the detection of Pebrine disease in silkworm eggs is critical. Current diagnostic methods can be timeconsuming and complex. This study aimed to develop a simplified and rapid diagnostic method using loop-mediated isothermal amplification (LAMP) technology to detect pebrine infection in silkworm mother moths. Eight primer candidates targeting the ribosomal gene region of microsporidia were designed and evaluated for specificity and detection sensitivity. A simplified nucleic acid extraction method was established, and isothermal amplification was performed using the selected primers. Of these, primers ID30 and ID45 showed no polymerization, while ID5, ID18, and ID76 exhibited nonspecific reactions, making them unsuitable. Primers ID1, ID6, ID45, and ID82 successfully amplified DNA only in the presence of pebrine, with ID82 demonstrating the best reproducibility and sensitivity, detecting as low as 2.5 pg/ul of DNA through electrophoresis and 5 pg/ul via a colorimetric change with phenol red. The entire process, from nucleic acid extraction to detection, was completed within 60 min. The use of the ID82 primer set in LAMP technology offers a promising and efficient approach for the rapid diagnosis of pebrine disease, potentially enhancing quality control in sericulture.

• Journal of Korean Society on Water Environment
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• v.40 no.1
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• pp.19-35
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• 2024

The global pandemic, coronavirus disease caused by Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has led to the implementation of wastewater surveillance as a means to monitor the spread of SARS-CoV-2 prevalence in the community. The challenging aspect of establishing wastewater surveillance requires a well-equipped laboratory for wastewater sample analysis. According to previous studies, RT-PCR-based molecular tests are the most widely used and popular detection method worldwide. However, this approach for the detection or quantification of SARS-CoV-2 from wastewater demands a specialized laboratory, skilled personnel, expensive instruments, and a workflow that typically takes 6 to 8 hours to provide results for a few samples. Rapid and reliable alternative detection methods are needed to enable less-well-qualified practitioners to set up and provide sensitive detection of SARS-CoV-2 within wastewater at regional laboratories. In some cases, the structural and molecular characteristics of SARS-CoV-2 are unknown, and various strategies for the correct diagnosis of COVID-19 have been proposed by research laboratories. The ongoing research and development of alternative and rapid technologies, namely RT-LAMP, ELISA, Biosensors, and GeneXpert, offer a wide range of potential options not only for SARS-CoV-2 detection but also for other viruses. This study aims to discuss the effective regional rapid detection and quantification methods in community wastewater.

• International journal of advanced smart convergence
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• v.8 no.1
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• pp.113-125
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• 2019

The radish skin and radish greens are an edible part of the radish. But they are removed before eating the radish and used as a byproduct or an animal feed material because of their tough and rough texture. This study was conducted to investigate the effect of supercritical heat-treated radish-extract on UV-induced HRM-2 wrinkled mouse animal model on anti-aging wrinkles. Supercritical heat-treated radish-extract was applied on the back of seven-weeks old HRM-2 mice. The effect of HRE on skin thickness, elasticity and wrinkle formation of the mice was observed by using UVB lamp to induce melanogenesis and wrinkle formation. As the result, increased depth of wrinkles was observed in the negative control group in comparison to the normal group. In contrast, decreased depth of wrinkles was observed in the radish-extract-free group compared to the negative control group. In the study of the effect of radish-extract on wrinkle-formation related gene expression and protein what protein expression, MMP-2 and MMP-9 gene expression significantly increased in the negative control group compared to the normal group. The gene expression reduced in dependence to the mass of radish-extract treated. Similar to quantitative results of mRNA expression, the expression of MMP-2 protein increased as a result of UVB-irradiation. The MMP-2 expression was inhibited in dependence to the mass of radish-extract treated. In conclusion, the supercritical heat-treated radish-extract has an effect on improving skin wrinkles not only when it is applied to the skin but also when orally ingested. Thus, it can be effectively used as a composition to health functional products. Therefore we can also conclude that radish a food that does not show any side-effects even upon long-term intake can reduce wrinkle formation as well as improve skin elasticity when taken regularly for a long period.

• Korean Journal of Poultry Science
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• v.39 no.3
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• pp.241-244
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• 2012

This study was performed to analyze the generational transmission and the expression of the foreign gene in the GFP transgenic chickens. The transmission rate and the expression of the GFP gene was investigated from the GFP transgenic rooster (G2) as the first founder to the ninth (G8). Analysis of GFP expression in hatched chickens was used the UV lamp. When GFP was expressed in the wings, bill and legs of a chick, the bird only was selected as a transgenic chick. The average transmission rate of the overall transgenic was 38~58%. These results showed that the transmission of the GFP gene in the transgenic chickens in accordance with the laws of Mendel's continues to the next generation without gene silencing.

• Journal of Plant Biotechnology
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• v.42 no.2
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• pp.104-110
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• 2015

We analyzed the effects of various LED light treatments (red 655 nm, blue 456 nm, white and mixed light) on growth pattern, gene expression and flavonoid contents in lettuce leaf. Plants treated with mixed light (red+blue+white) showed better growth performance than those treated with single LED and fluorescent lamp (FL). Expression analysis of the eight genes involved in flavonoid biosynthesis in plants treated with LED light was examined. Results showed that red lettuce grown under mixed light showed high expression levels of LsC4H, LsF3H and LsDRF genes. Morever, the same treatment plants possessed higher content of gallic acid, chlorogenic acid and quercetin contents than those in plants exposed to single light. However, the highest total anthocyanin content was identified in plants treated with red+blue light and the lowest content was identified in plants exposed to white fluorescent lamp and single LED light condition. Thus, this study indicates that the ratio of blue to red LEDs is important for the morphology, growth, and phenolic compounds with anthocyanin properties in the two lettuce cultivars tested.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.5 no.1
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• pp.57-64
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• 1969

Several field and sweet corn varietiea from several sources were crossed with a variety carrying the opaque-2 gene to determine the phenotypic interactions in the breeding of high lysine maize. Although opaque-2 lines showed lower protein content than the corresponding normal varieties, there was no correlation between the protein levels of the two types. opaque-2 maize contained more lysine, but no relationship was found between the protein content and the lysine content of either normal or opaque-2 types, suggesting that high lysine corn using the opaque-2 gene may be developed independently from the protein content. The F2 segregation ratios for normal and opaque-2, 100-kernel weights, percentage seed set, opaque-2 phenotype, disease susceptibility, and the relationship between protein and lysine content of normal and opaque-2 were investigated. The determinations and observations were made on the F2, F3, and BC1 Lysine content was determined by the ion exchangeresin combined with paper chromatography method. Most crosses segregated in a 1-opaque-2 : 3-normal ratio as expected. Opaque-2 segregates were lighter than the normal type and smaller in size. A mottled phenotype of opaque-2 maize observed in the Philippines yellow endersperm. In some varieties opaque-2 maize was very susceptible to the ear and kernel rot disease. No. 5(female) and opaque-2(male). Selectlon of a double mutant of waxy and opaque-2 by using the iodine technique and electric lamp was discussed. opaque-2 and floruy-2 were not allels. Different percentage of seed set were observed in the segregation of aewx crossed with opaque-2. An unusual gametophytie relationship was involved in a cross between Glutinous.

• Reproductive and Developmental Biology
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• v.37 no.4
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• pp.269-279
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• 2013

Low efficiency of somatic cell nuclear transfer (SCNT) is attributed to incomplete reprogramming of transfered nuclei into oocytes. Trichostatin A (TSA), histone deacetylase inhibitor and 5-aza-2'deoxycytidine (5-aza-dC), DNA methylation inhibitor has been used to enhance nuclear reprogramming following SCNT. However, it was not known molecular mechanism by which TSA and 5-aza-dC improve preimplantation embryo and fetal development following SCNT. The present study investigates embryo viability and gene expression of cloned porcine preimplantation embryos in the presence and absence of TSA and 5-aza-dC as compared to embryos produced by parthenogenetic activation. Our results indicated that TSA treatment significantly improved development. However 5-aza-dC did not improve development. Presence of TSA and 5-aza-dC significantly improved total cell number, and also decreased the apoptotic and autophagic index. Three apoptotic-related genes, Bak, Bcl-xL, and Caspase 3 (Casp3), and three autophagic-related genes, ATG6, ATG8, and lysosomal-associated membrane protein 2 (LAMP2), were measured by real time RT-PCR. TSA and 5-aza-dC treatment resulted in high expression of anti-apoptotic gene Bcl-xL and low pro-apoptotic gene Bak expression compared to untreated NT embryos or parthenotes. Furthermore, LC3 protein expression was lower in NT-TSA and NT-5-aza-dC embryos than those of NT and parthenotes. In addition, TSA and 5-aza-dC treated embryos displayed a global acetylated histone H3 at lysine 9 and methylated DNA H3 at lysine 9 profile similar to the parthenogenetic blastocysts. Finally, we determined that several DNA methyltransferase genes Dnmt1, Dnmt3a and Dnmt3b. NT blastocysts showed higher levels Dnmt1 than those of the TSA and 5-aza-dC blastocysts. Dnmt3a is lower in 5-aza-dC than NT, NTTSA and parthenotes. However, Dnmt3b is higher in 5-aza-dC than NT and NTTSA. These results suggest that TSA and 5-aza-dC positively regulates nuclear reprogramming which result in modulation of apoptosis and autophagy related gene expression and then reduce apoptosis and autophagy. In addition, TSA and 5-aza-dC affects the acetylated and methylated status of the H3K9.