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Development of TaqMan Quantitative PCR Assays for Duplex Detection of Dirofilaria immitis COI and Dog GAPDH from Infected Dog Blood (심장사상충에 감염된 개 혈액에서 Dirofilaria immitis의 COI와 개의 GAPDH를 이중 검출하기 위한 정량적 TaqMan PCR 분석법의 개발)

  • Oh, In Young;Kim, Kyung Tae;Gwon, Sun-Yeong;Sung, Ho Joong
    • Korean Journal of Clinical Laboratory Science
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    • v.51 no.1
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    • pp.64-70
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    • 2019
  • Dirofilaria immitis (D. immitis) is a filarial nematode that causes cardiopulmonary dirofilariasis in dogs. In the late stages of infection, infected dogs show one or more symptoms and advanced heart disorder with perivascular inflammation. To detect D. immitis specifically and efficiently in the early stages of infection, a duplex TaqMan qPCR assay was developed based on previous studies using primers and probes specialized to detect D. immitis cytochrome c oxidase subunit I (COI) and dog glyceraldehyde-3-phosphate dehydrogenase (GAPDH). As positive controls, plasmid DNAs were constructed from D. immitis COI or dog GAPDH and a TA-cloning vector. Simplex and duplex TaqMan qPCR assays were performed using the specific primers, probes, and genomic or plasmid DNA. The duplex reaction developed could detect D. immitis COI and dog GAPDH in the same sample simultaneously after optimization of the primer concentrations. The limit of detection was 25 copies for the simplex and duplex assays, and both showed good linearity, high sensitivity, and excellent PCR efficiency. The duplex assays for pathogen detection reduce the costs, labor, and time compared to simplex reactions. Therefore, the duplex TaqMan qPCR assay developed herein will allow efficient D. immitis detection and quantification from a large number of samples simultaneously.

Spatial and seasonal distributions of the phototrophic dinoflagellate Biecheleriopsis adriatica (Suessiaceae) in Korea: quantification using qPCR

  • Kang, Hee Chang;Jeong, Hae Jin;Ok, Jin Hee;You, Ji Hyun;Jang, Se Hyeon;Lee, Sung Yeon;Lee, Kyung Ha;Park, Jae Yeon;Rho, Jung-Rae
    • ALGAE
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    • v.34 no.2
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    • pp.111-126
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    • 2019
  • The phototrophic dinoflagellate Biecheleriopsis adriatica is a small suessioid species characterized by a fragile thin wall. Although the morphology of this dinoflagellate is well established, there is currently little information available on its distribution and the environmental factors that influence this distribution. Thus, to investigate the spatial and seasonal distributions of the vegetative cells of B. adriatica in Korean waters, surface water samples were collected on a seasonal basis from 28 stations in the East, West, and South Sea of Korea and Jeju Island from April 2015 to October 2018, and abundances of the vegetative cells of B. adriatica were quantified using quantitative real-time polymerase chain reactions, for which we developed the species-specific primer and probe set. Simultaneously, major environmental parameters, including temperature, salinity, nutrient concentrations, and dissolved oxygen concentrations were measured. The vegetative cells of B. adriatica were detected at 20 of the 28 sampling stations: 19 stations in summer and 6 in autumn, although from no stations in either spring or winter. The ranges of water temperature and salinity at sites where this species was detected were $17.7-26.4^{\circ}C$ and 9.9-34.3, respectively, whereas those of nitrate and phosphate concentrations were not detectable-96.2 and $0.18-2.66{\mu}M$, respectively. Thus, the sites at which this species is found are characterized by a narrow range of temperature, but wide ranges of salinity and concentrations of nitrate and phosphate. The highest abundance of the vegetative cells of B. adriatica was $41.7cells\;mL^{-1}$, which was recorded in Jinhae Bay in July 2018. In Jinhae Bay, the abundance of vegetative cells was significantly positively correlated with the concentration of nitrate, but was negatively correlated with salinity. On the basis of these findings, it appears that the abundance of B. adriatica vegetative cells shows strong seasonality, and in Jinhae Bay, could be affected by the concentrations of nitrate.

Comparison of the spatial-temporal distributions of the heterotrophic dinoflagellates Gyrodinium dominans, G. jinhaense, and G. moestrupii in Korean coastal waters

  • Lee, Sung Yeon;Jeong, Hae Jin;Kang, Hee Chang;Ok, Jin Hee;You, Ji Hyun;Park, Sang Ah;Eom, Se Hee
    • ALGAE
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    • v.36 no.1
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    • pp.37-50
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    • 2021
  • Heterotrophic dinoflagellates Gyrodinium spp. are one of the major grazers of phytoplankton in many coastal waters. Gyrodinium dominans, G. jinhaense, and G. moestrupii have similar morphologies but different edible prey species. To explore the variations in the ecological niches of these three species, we investigated their spatial-temporal distributions in Korean waters. Because of the high similarity in morphology among these three Gyrodinium species, we used real-time polymerase chain reactions to quantify their abundance in water samples that were seasonally collected from 28 stations along the Korean Peninsula from April 2015 to October 2018. Cells of G. dominans were found at all sampling stations, G. jinhaense at 26 stations, and G. moestrupii at 22 stations, indicating that all three species were widely distributed in Korea. Furthermore, all three species displayed strong seasonal distributions. The largest numbers of the stations where G. dominans and G. jinhaense cells were present were found during the summer (26 and 23 stations, respectively), but that for G. moestrupii was found in the autumn (15 stations). The abundance of G. dominans was positively correlated with that of G. jinhaense, but not with that of G. moestrupii. The highest abundances of G. dominans (202.5 cells mL-1) and G. jinhaense (20.2 cells mL-1) were much greater than that of G. moestrupii (1.2 cells mL-1). The highest abundances of G. dominans and G. jinhaense were found in July, whereas that of G. moestrupii was found in March. The abundances of G. dominans and G. jinhaense, but not G. moestrupii, were positively correlated with water temperature. Therefore, the spatial-temporal distributions of G. dominans and G. jinhaense were closer than those of G. moestrupii and G. dominans or G. jinhaense. This differs from results based on the relative differences in ribosomal DNA sequences and the types of edible prey reported in the literature. Thus, the variations in spatial-temporal distributions and prey species of these three Gyrodinium species suggest that they may have different ecological niches in Korean coastal waters.

Analysis of Volatile Organic Compounds in Sediments Using HS-GC/MS - Confirmation of Matrix Effects in External and Internal Standard Methods - (HS-GC/MS를 이용한 퇴적물 중 휘발성유기화합물 분석 - 외부 및 내부표준방법에서 매질영향 확인 -)

  • Shin, Myoung-Chul;Jung, Da-som;Noh, Hye-ran;Yu, Soon-ju;Seo, Yong-Chan;Lee, Bo-Mi
    • Journal of Korean Society on Water Environment
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    • v.37 no.6
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    • pp.510-519
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    • 2021
  • Volatile Organic Compounds (VOCs) in sediments, which can cause human health problems, have been monitored in Korea since 2014. Measured VOC concentrations can be affected by matrix type and the volatility of target substances. In this study, (1) VOCs volatility and the influence of matrix interference were confirmed, and (2) internal standards (IS) method was applied to improve analytical method. For these purposes, method detection limit (MDL), calibration linearity, precision and accuracy of VOCs were compared in various matrices using the IS. Some of VOCs in sediments showed different peak areas and reduced rates compared to water matrix. It was suggested that adsorption properties of sediments hindered the migration to vapor during heat pretreatment in headspace method. A calibration curve was created in clean sand. Recovery rates for the calibration curve method and IS applying method were 64.1~83.1% and 99.1~119.3%, respectively. Relative standard deviations ranged from 11.1% to 21.6% for the calibration curve method and those for IS ranged 4.7% to 13.7%. In case of real sediment, calibration curve and 1,2-Dichlorobenzene-d4 (ODCB) among IS were not suitable. The average recovery rate of Fluorobenzene (FBZ) increased by 56.4% and Relative Standard Deviation (RSD) by 4.7%. However, the recovery rate was increased in the samples with large values of igniting intensity. This study confirmed that influence of the matrix of VOCs in sediment, and addition of IS materials improved precision and accuracy. Although IS corrects volatilization and adsorption, it is recommended that more than two types of IS should be added rather than single.

The Effect of Social Function and Telepresence on Intention to Offer Support Through Trust of Metaverse Participants (메타버스의 사회적 기능과 원격실재감이 메타버스 참여 주체의 신뢰를 통해 요청지원 의도에 미치는 영향)

  • Hwang, Inho
    • Journal of Korea Society of Industrial Information Systems
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    • v.27 no.3
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    • pp.29-46
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    • 2022
  • COVID-19 has radically changed the behavior of members of society for exchange. In particular, the strong contagiousness of the virus is increasing networking on online platforms while reducing people's networking in the real world. Recently, the metaverse, which strengthened the presence based on 3D technology, is attracting attention from members of society such as individuals and companies. We present a method to improve metaverse utilization from the perspective of organizations and employees who have introduced metaverse for work. In other words, we check the effect of metaverse social function and telepresence on the employee's intention to offer support by improving the trust of the metaverse participants. We obtained samples through questionnaires targeting employees of organizations that introduced metaverse to their work, and verified the research hypothesis by applying the structural equation model. As a result, social interactivity, reciprocal favor, and telepresence of metaverse partially affected metaverse trust (platform, peer, organization), and metaverse trust increased the intention to offer support. Our study suggests a strategic direction to improve the metaverse utilization and exchange level of employees of organizations who want to use the metaverse for business.

Maternal nutrition altered embryonic MYOD1, MYF5, and MYF6 gene expression in genetically fat and lean lines of chickens

  • Li, Feng;Yang, Chunxu;Xie, Yingjie;Gao, Xiang;Zhang, Yuanyuan;Ning, Hangyi;Liu, Guangtao;Chen, Zhihui;Shan, Anshan
    • Animal Bioscience
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    • v.35 no.8
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    • pp.1223-1234
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    • 2022
  • Objective: The objectives of this study were to evaluate the effects of daily feed intake during the laying period on embryonic myogenic differentiation 1 (MYOD1), myogenic factor 5 (MYF5), and myogenic factor 6 (MYF6) gene expression in genetically fat and lean lines of chickens. Methods: An experiment in a 2×2 factorial design was conducted with two dietary intake levels (100% and 75% of nutrition recommendation) and two broiler chicken lines (fat and lean). Two lines of hens (n = 384 for each line) at 23th week of age were randomly divided into 4 treatments with 12 replicates of 16 birds. The experiment started at 27th week of age (5% egg rate) and ended at 54th week of age. Hatched eggs from the medium laying period were collected. Real time polymerase chain reaction analysis was used to analyse the MYOD1, MYF5, and MYF6 mRNA levels of E7, E9, E11, E13, and E15 body tissues and E17, E19, and E21 chest and thigh muscle samples. Results: The results indicated that there were significant effects of line, dietary intake, and interactions between them on MYOD1, MYF5, and MYF6 gene mRNA expression levels in embryonic tissues. Low daily feed intake did not change the expression trend of MYOD1 mRNA in either line, but changed the peak values, especially in lean line. Low daily feed intake altered the trend in MYF5 mRNA expression level in both lines and apparently delayed its onset. There was no apparent effect of low daily feed intake on the trends of MYF6 mRNA expression levels in either line, but it significantly changed the values on many embryonic days. Conclusion: Maternal nutrient restriction affects myogenesis and is manifested in the expression of embryonic MYOD1, MYF5, and MYF6 genes. Long term selection for fat deposition in broiler chickens changes the pattern and intensity of myogenesis.

EID3 Promotes Glioma Cell Proliferation and Survival by Inactivating AMPKα1

  • Xiang, Yaoxian;Zhu, Lei;He, Zijian;Xu, Lei;Mao, Yuhang;Jiang, Junjian;Xu, Jianguang
    • Journal of Korean Neurosurgical Society
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    • v.65 no.6
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    • pp.790-800
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    • 2022
  • Objective : EID3 (EP300-interacting inhibitor of differentiation) was identified as a novel member of EID family and plays a pivotal role in colorectal cancer development. However, its role in glioma remained elusive. In current study, we identified EID3 as a novel oncogenic molecule in human glioma and is critical for glioma cell survival, proliferation and invasion. Methods : A total of five patients with glioma were recruited in present study and fresh glioma samples were removed from patients. Four weeks old male non-obese diabetic severe combined immune deficiency (NOD/SCID) mice were used as transplant recipient models. The subcutaneous tumor size was calculated and recorded every week with vernier caliper. EID3 and AMP-activated protein kinase α1 (AMPKα1) expression levels were confirmed by real-time polymerase chain reaction and Western blot assays. Colony formation assays were performed to evaluate cell proliferation. Methyl thiazolyl tetrazolium (MTT) assays were performed for cell viability assessment. Trypan blue staining approach was applied for cell death assessment. Cell Apoptosis DNA ELISA Detection Kit was used for apoptosis assessment. Results : EID3 was preferentially expressed in glioma tissues/cells, while undetectable in astrocytes, neuronal cells, or normal brain tissues. EID3 knocking down significantly hindered glioma cell proliferation and invasion, as well as induced reduction of cell viability, apoptosis and cell death. EID3 knocking down also greatly inhibited tumor growth in SCID mice. Knocking down of AMPKα1 could effectively rescue glioma cells from apoptosis and cell death caused by EID3 absence, indicating that AMPKα1 acted as a key downstream regulator of EID3 and mediated suppression effects caused by EID3 knocking down inhibition. These findings were confirmed in glioma cells generated patient-derived xenograft models. AMPKα1 protein levels were affected by MG132 treatment in glioma, which suggested EID3 might down regulate AMPKα1 through protein degradation. Conclusion : Collectively, our study demonstrated that EID3 promoted glioma cell proliferation and survival by inhibiting AMPKα1 expression. Targeting EID3 might represent a promising strategy for treating glioma.

Dietary Iron Intake of Koreans Estimated using 2 Different Sources of Iron Contents are Comparable: Food & Nutrient Database and Iron Contents of Cooked Foods in the Korean Total Diet Study (우리 국민의 철 섭취량 평가: 식품별 영양성분 함량자료와 한국형 총식이조사 기반 추정량 비교)

  • Lee, Jeeyeon;Kwon, Sung Ok;Yeoh, Yoonjae;Seo, Min Jeong;Lee, Gae Ho;Kim, Cho-il
    • Korean Journal of Community Nutrition
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    • v.27 no.3
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    • pp.245-253
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    • 2022
  • Objectives: This study was conducted to find out if the dietary iron intake of Koreans estimated by 2 different methods (iron content sources) using the food intake data from the Korea National Health and Nutrition Examination Survey (KNHANES) are comparable. One method was based on the KNHANES's Food & Nutrient Database (FND) derived mainly from the Korean Food Composition Table and the other used the iron content (IC) of food samples processed in the Korean Total Diet Study (KTDS). Methods: Dietary intake data from the 2013-2016 KNHANES was used to select representative foods (RFs) in KTDS for iron analysis. Selection of the RFs and cooking methods for each RF (RF × cooking method pair) was performed according to the 'Guidebook for Korean Total Diet Studies' and resulted in a total of 132 RFs and 224 'RF × cooking method' pairs. RFs were collected in 9 metropolitan cities nationwide once or twice (for those with seasonality) in 2018 and made into 6 composites each, based on the origin and season prior to cooking. Then, the RF composites prepared to a 'table ready' state for KTDS were analyzed by Inductively Coupled Plasma-Mass Spectrometry (ICP-MS). Dietary iron intake of the Korean population was estimated using only RFs' intake data based on the 2 sources of iron content, namely FND-KNHANES and IC-KTDS. Results: RFs in KTDS covered 92.0% of total food intake of Koreans in the 2016-2018 KNHANES. Mean iron intake of Koreans was 7.77 mg/person/day by IC-KTDS vs 9.73 mg/person/day by FND-KNHANES. The major food groups contributing to iron intake were meats (21.7%), vegetables (20.5%), and grains & cereals (13.4%) as per IC-KTDS. On the other hand, the latter source (FND-KNHANES) resulted in a very different profile: grains & cereals (31.1%), vegetables (16.8%), and meats (15.3%). While the top iron source was beef, accounting for 8.6% in the former, it was polished rice (19.2%) in the latter. There was a 10-fold difference in the iron content of polished rice between 2 sources that iron intakes excluding the contribution by polished rice resulted in very similar values: 7.58 mg/person/day by IC-KTDS and 7.86 mg/person/day by FND-KNHANES. Conclusions: This study revealed that the dietary iron intake estimated by 2 different methods were quite comparable, excluding one RF, namely polished rice. KTDS was thus proven to be a useful tool in estimating a 'closer-to-real' dietary intake of nutrients for Koreans and further research on various nutrients is warranted.

Objective Evidence for the Effectiveness of Single-session Treatment with a Spinal Thermal Massage Device: A Pilot Study (척추온열마사지기기의 1회 치료의 효과에 대한 객관적 증거: 선행 연구)

  • Na, Yeong-Il;Kim, Si-Yun;Baek, Seung-Min;Lee, Jong-Hoo
    • Journal of Industrial Convergence
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    • v.20 no.10
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    • pp.209-218
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    • 2022
  • Individuals often report significant relief from pain and stress even after a single session of massage therapy; however, no previous studies have provided objective evidence supporting the effectiveness of a solitary massage therapy session. In the present study, we aimed to investigate the effectiveness of one-time treatment with a spinal thermal massage device reported to exert the same therapeutic effects as massage therapy in terms of pain reduction and stress relief. A man with chronic low back pain (LBP) underwent two rounds of experiments involving spinal massage treatment and bed rest, respectively. Pain was assessed using a visual analog scale, and heart rate variability (HRV) was measured in real-time to examine autonomic nervous system (ANS) activity. Blood samples were obtained at five points during each round of the experiment to examine changes in cortisol, epinephrine, and norepinephrine. Spinal massage significantly reduced pain and enhanced parasympathetic activity when compared with the bed rest condition. In addition, both epinephrine and norepinephrine levels were lower following spinal massage than following bed rest. These results are consistent with the reported effects of conventional massage therapy and support the effectiveness of one-time treatment using a spinal thermal massage device.

Molecular Characterization of Vibrio parahaemolyticus Isolates from Seawater, Fish Tanks, and Distributed Fishery Products in Jeju (제주지역 해수, 수족관물, 유통수산물에서 분리된 장염비브리오균의 분자생물학적 특성)

  • Man Jae Cho;Eunok Kang;Ye-seul Heo;Eun A Koh
    • Journal of Food Hygiene and Safety
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    • v.38 no.4
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    • pp.246-254
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    • 2023
  • V. parahaemolyticus causes waterborne and foodborne disease such as acute diarrhea. In this study, V. parahaemolyticus isolates from seawater, fish tanks, and distributed fishery products in Jeju were investigated for potential toxin or species-specific genes (tdh, trh, tlh, and toxR) using RT-PCR and their genetic characteristics were analyzed using Pulsed-field gel electrophoresis (PFGE). Overall, V. parahaemolyticus of 90 strains (36.7%), including 33 strains from seawater, 8 strains from fish tanks, and 50 strains from fishery products, were isolated from 245 samples. All V. parahaemolyticus strains did not detect the tdh gene, whereas all strains detected tlh or toxR genes. In addition, trh genes were detected in 3 strains from seawater and 1 strain from fishery products. Monthly quantitative testing of seawater revealed that V. parahaemolyticus was positively correlated with water temperature. The 90 strains of V. parahemolyticus obtained in this study showed by gene homology between types, ranging from 64.0-97.3%. Among these, thirteen types showed 100% homology between genes. These results indicate that continuous monitoring is needed to facilitate food poisoning epidemiological investigations because some isolated V. parahaemolyticus strains harbored toxin genes and V. parahaemolyticus strains isolated from seawater, fish tanks, and distributed fishery products showed genetic similarity.