• Parasites, Hosts and Diseases
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• v.56 no.5
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• pp.419-427
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• 2018

This study aimed to develop a new multiplex real-time PCR detection method for 3 species of waterborne protozoan parasites (Cryptosporidium parvum, Giardia lamblia, and Cyclospora cayetanensis) identified as major causes of traveler's diarrhea. Three target genes were specifically and simultaneously detected by the TaqMan probe method for multiple parasitic infection cases, including Cryptosporidium oocyst wall protein for C. parvum, glutamate dehydrogenase for G. lamblia, and internal transcribed spacer 1 for C. cayetanensis. Gene product 21 for bacteriophage T4 was used as an internal control DNA target for monitoring human stool DNA amplification. TaqMan probes were prepared using 4 fluorescent dyes, $FAM^{TM}$, $HEX^{TM}$, $Cy5^{TM}$, and CAL Fluor $Red^{(R)}$ 610 on C. parvum, G. lamblia, C. cayetanensis, and bacteriophage T4, respectively. We developed a novel primer-probe set for each parasite, a primer-probe cocktail (a mixture of primers and probes for the parasites and the internal control) for multiplex real-time PCR analysis, and a protocol for this detection method. Multiplex real-time PCR with the primer-probe cocktail successfully and specifically detected the target genes of C. parvum, G. lamblia, and C. cayetanensis in the mixed spiked human stool sample. The limit of detection for our assay was $2{\times}10$ copies for C. parvum and for C. cayetanensis, while it was $2{\times}10^3$ copies for G. lamblia. We propose that the multiplex real-time PCR detection method developed here is a useful method for simultaneously diagnosing the most common causative protozoa in traveler's diarrhea.

• Microbiology and Biotechnology Letters
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• v.32 no.3
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• pp.230-237
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• 2004

In this study nitroreductase from Pseudomonas sp. HK-6 capable of degrading 2,4,6-trinitrotoluene (TNT) was characterized. Through a series of purification process including ammonium sulfate precipitation, DEAE-sepharose, and Q-sepharose, three different fractions I, II, and III having the enzyme activity of NTRs whose molecular weights were approximately 27 kDa were detected in fractions from HK-6 cells. Specific activity of the three fractions were approximately 4.85 unit/mg, 5.47 unit/mg, and 5.01 unit/mg, and concentrated to 9.0-, 10.1-, and 9.3-fold compared to crude extract, respectively. The optimal pH and temperature for the three NTR fractions were approximately 7.5 and $30^{\circ}C$, respectively. Metal ions, $Ag^{＋}$ , $Cu^{ 2+}$, $Hg^{2＋}$ inhibited approximately 70% of enzymes activities of all NTR, while $Fe^{2＋}$ did not stimulate or inhibit the activities. Monitoring the effect of chemicals on the enzyme activity revealed that those NTR fractions lost enzyme activity in presence of $\beta$-mercaptoethanol, but were a little influenced by dithiothreitol, EDTA and NaCl. The three NTR fractions demonstrated enzyme activities for nitrobenzene and RDX as well as TNT.

• 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.

• Asian-Australasian Journal of Animal Sciences
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• v.28 no.11
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• pp.1649-1656
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• 2015

Gene expression profiling has offered new insights into postmortem molecular changes associated with meat quality. To acquire reliable transcript quantification, high quality RNA is required. The objective of this study was to analyze integrity of RNA isolated from chicken skeletal muscle (pectoralis major) and its capability of serving as the template in quantitative real-time polymerase chain reaction (qPCR) as a function of postmortem intervals representing the end-points of evisceration, carcass chilling and aging stages in chicken abattoirs. Chicken breast muscle was dissected from the carcasses (n = 6) immediately after evisceration, and one-third of each sample was instantly snap-frozen and labeled as 20 min postmortem. The remaining muscle was stored on ice until the next rounds of sample collection (1.5 h and 6 h postmortem). The delayed postmortem duration did not significantly affect $A_{260}/A_{280}$ and $A_{260}/A_{230}$ ($p{\geq}0.05$), suggesting no altered purity of total RNA. Apart from a slight decrease in the 28s:18s ribosomal RNA ratio in 1.5 h samples (p<0.05), the value was not statistically different between 20 min and 6 h samples ($p{\geq}0.05$), indicating intact total RNA up to 6 h. Abundance of reference genes encoding beta-actin (ACTB), glyceraldehyde 3-phosphate dehydrogenase (GAPDH), hypoxanthine-guanine phosphoribosyltransferase (HPRT), peptidylprolylisomerase A (PPIA) and TATA box-binding protein (TBP) as well as meat-quality associated genes (insulin-like growth factor 1 (IGF1), pyruvate dehydrogenase kinase isozyme 4 (PDK4), and peroxisome proliferator-activated receptor delta (PPARD) were investigated using qPCR. Transcript abundances of ACTB, GAPDH, HPRT, and PPIA were significantly different among all postmortem time points (p<0.05). Transcript levels of PDK4 and PPARD were significantly reduced in the 6 h samples (p<0.05). The findings suggest an adverse effect of a prolonged postmortem duration on reliability of transcript quantification in chicken skeletal muscle. For the best RNA quality, chicken skeletal muscle should be immediately collected after evisceration or within 20 min postmortem, and rapidly preserved by deep freezing.

• Asian-Australasian Journal of Animal Sciences
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• v.31 no.2
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• pp.167-179
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• 2018

Objective: The aim of this study was to create a set of microsatellite markers with high polymorphism for the genetic monitoring and genetic structure analysis of local goose populations. Methods: Novel microsatellite markers were isolated from the genomic DNA of white Roman geese using short tandem repeated probes. The DNA segments, including short tandem repeats, were tested for their variability among four populations of geese from the Changhua Animal Propagation Station (CAPS). The selected microsatellite markers could then be used to monitor genetic variability and study the genetic structures of geese from local geese farms. Results: 14 novel microsatellite loci were isolated. In addition to seven known loci, two multiplex sets were constructed for the detection of genetic variations in geese populations. The average of allele number, the effective number of alleles, the observed heterozygosity, the expected heterozygosity, and the polymorphism information content were 11.09, 5.145, 0.499, 0.745, and 0.705, respectively. The results of analysis of molecular variance and principal component analysis indicated a contracting white Roman cluster and a spreading Chinese cluster. In white Roman populations, the CAPS populations were depleted to roughly two clusters when K was set equal to 6 in the Bayesian cluster analysis. The founders of private farm populations had a similar genetic structure. Among the Chinese geese populations, the CAPS populations and private populations represented different clads of the phylogenetic tree and individuals from the private populations had uneven genetic characteristics according to various analyses. Conclusion: Based on this study's analyses, we suggest that the CAPS should institute a proper breeding strategy for white Roman geese to avoid further clustering. In addition, for preservation and stable quality, the Chinese geese in the CAPS and the aforementioned proper breeding scheme should be introduced to geese breeders.

• Journal of Korean Society of Forest Science
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• v.107 no.2
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• pp.151-157
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• 2018

This study was to analyze the genetic diversity of Quercus gilva Blume growing in Jeju Island for developing a preservation strategy. We examined the genetic diversity and structure using 6 ISSR primers and investigated 67 polymorphic ISSR amplicons in 80 trees distributed among five populations. The average of proportion of polymorphic loci were 93%, the average level of Shannon's information index was 0.237, and Nei's genetic diversity was 0.156. According to the analysis of the molecular variance (AMOVA), $F_{st}$ was 0.169 indicating there was a genetic variation among five populations. 17% of the total variation was allocated among the five populations, while the other 83% of the total variation was in individual trees in each population. The result could be due to the uneven number of trees among the five populations. Based on these results, the preservation strategy could be developed, for examples, considering for designation as "forest genetic resources conservation area" about the habitat, monitoring continuously, fostering the growth of seedling, ex situ preservation of genetic resources, and comparing the differences of environmental and genetic characteristic with population in ex situ.

• Journal of Life Science
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• v.27 no.3
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• pp.301-309
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• 2017

This study is examined river naturality and vegetative composition of river riparian zones to identify their most important sources of variation. Information on plant species cover and on physical characteristics that occur at upper, medium, and low areas was collected for 30 riparian plots located throughout the Youngjeong River in Korea. The riparian areas of river banks are dominated by mixed sediment and the vegetation is composed of herbs, shrub, and trees. The floristic characterization of riparian at this river during 2015 season was identified with a total of 28 families, 72 genera, 75 species, 13 varieties, 23 associations. The vegetations of low water's edge and flood way at upper region were naturally formed various vegetation communities by natural erosion. Forty plant species were identified around the upper region, where the dominant growth form was mostly trees. The flood way vegetation at middle region was both of natural vegetation and artificial vegetation. Land uses in riparian zones river levee at low region were bush or grassland as natural floodplain. The values of cover-abundance at upper, middle, and low region were total 9.26, 7.24, and 7.56, respectively. Grasses and forbs at the Youngjeong River have similar cover-abundance values. Recent, many riparian areas of this river have been lost or degraded for commercial and industrial developments. Thus, monitoring for biological diversity of plant species of this river is necessary for an adaptive management approach and the successful implementation of ecosystem management.

• Pediatric Infection and Vaccine
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• v.27 no.2
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• pp.90-101
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• 2020

Purpose: The multiplex reverse transcriptase-polymerase chain reaction (RT-PCR) test developed recently can help detect enteric pathogens of acute gastroenteritis (AGE). This study aimed to investigate the epidemiology of pathogens in children with AGE using the multiplex RT-PCR. Methods: From May 2015 to June 2018, multiplex RT-PCR tests were performed to identify pathogens in the feces of pediatric patients diagnosed with AGE at a secondary hospital in Seoul, Korea. Results: Of the 1,366 stool samples examined for viral pathogens, 483 (35.3%) tested positive for ≥1 pathogen. Group A rotavirus (RV) was detected in 106 cases (7.8%). The positivity rate increased annually from 3.0% (8/263) to 16.7% (48/288) and surged in 2018 (P<0.001). Norovirus (NoV) GII was the most common viral pathogen (263/1,366, 19.3%), and the positivity rate did not increase during the 3 years. Of the 304 stool samples tested for bacterial pathogens, Campylobacter spp. was the most common bacterial pathogen (32/304, 10.5%), followed by Clostridium difficile (22/304, 7.2%) and Salmonella spp. (17/304, 5.6%). The positivity rate of these bacterial pathogens did not change significantly during the study period. Conclusions: NoV GII is the main pathogen in childhood AGE since the introduction of RV vaccine, yet the number of rotavirus-infected patients increased during our study, especially in 2018. Therefore, further research is needed including the possibility of emergence of novel RV strains. Campylobacter spp. is the predominant cause of bacterial AGE in children. For proper treatment, the clinical characteristics of the bacteria should be taken into consideration, and continuous monitoring is necessary.

• Nuclear Medicine and Molecular Imaging
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• v.42 no.2
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• pp.153-163
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• 2008

Medical imaging modalities to image either anatomical structure or functional processes have developed along somewhat independent paths. Functional images with single photon emission computed tomography (SPECT) and positron emission tomography (PET) are playing an increasingly important role in the diagnosis and staging of malignant disease, image-guided therapy planning, and treatment monitoring. SPECT and PET complement the more conventional anatomic imaging modalities of computed tomography (CT) and magnetic resonance (MR) imaging. When the functional imaging modality was combined with the anatomic imaging modality, the multimodality can help both identify and localize functional abnormalities. Combining PET with a high-resolution anatomical imaging modality such as CT can resolve the localization issue as long as the images from the two modalities are accurately coregistered. Software-based registration techniques have difficulty accounting for differences in patient positioning and involuntary movement of internal organs, often necessitating labor-intensive nonlinear mapping that may not converge to a satisfactory result. These challenges have recently been addressed by the introduction of the combined PET/CT scanner and SPECT/CT scanner, a hardware-oriented approach to image fusion. Combined PET/CT and SPECT/CT devices are playing an increasingly important role in the diagnosis and staging of human disease. The paper will review the development of multi modality instrumentations for clinical use from conception to present-day technology and the application software.

• Korean Journal of Plant Taxonomy
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• v.43 no.4
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• pp.267-273
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• 2013

This study employed inter-simple-sequence repeat (ISSR) to assess genetic variation among 189 individuals representing 10 populations (nine in Korea and one in Japan) of Millettia japonica, which has recently been lifted from the endangered species of Korea. The calculated Shannon's information index value (I = 0.2689) of the species was appreciable and was higher than other endangered leguminous woody taxa. Gochang (I = 0.2968), Namhae (I = 0.2951), and Mt. Toham (I = 0.2823) populations showed relatively high genetic diversity, whereas the Kyushu (in Japan) population (I = 0.2487) exhibited the lowest. The results of an analysis of molecular variance indicated that 86.49% of the diversity was attributed to within populations, and 13.51% to differences among populations, suggesting that M. japonica populations do not have significant geographic differentiation and that the gene flow between populations exists to some extent (Nm = 1.8446). Continuous habitat monitoring should be conducted to conserve genetic diversity of M. japonica, particularly for those populations with relatively high genetic diversity. Selection of many individuals from the populations in Gochang, Namhae, and Mt. Toham is thought to be an appropriate strategy for ex situ conservation of M. japonica in Korea.