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Molecular Characterization and Expression Analysis of Nucleoporin 210 (Nup210) in Chicken

  • Ndimukaga, Marc;Bigirwa, Godfrey;Lee, Seokhyun;Lee, Raham;Oh, Jae-Don
    • Korean Journal of Poultry Science
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    • v.46 no.3
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    • pp.185-191
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    • 2019
  • Nucleoporin 210 (Nup210) is associated with several physiological processes including muscle and neural cell differentiation, autoimmune diseases, and peripheral T cell homeostasis. Chicken Nup210 (chNup210) gene was originally identified as one of the differentially expressed genes (DEGs) in the kidney tissues of chicken. To elucidate the role of Nup210 in metabolic disease of chicken, we studied the molecular characteristics of chNup210 and analyzed its gene expression under the stimulation of Toll-like receptor 3 (TLR3) ligands. The Nup210 genomic DNA and amino acid sequences of various species including fowls, fishes, and mammals were retrieved from the Ensemble database and subjected to bioinformatics analyses. The expression of Nup210 from several chicken tissues was probed through qRT-PCR, and chicken fibroblast DF-1 cell line was used to determine the change in expression of chNup210 after stimulation with TLR3 ligand, polyinosinic-polycytidylic acid (poly (I:C)). The chNup210 gene was highly expressed in chicken lung and spleen tissues. Although highly conserved among the species, chNup210 was evolutionary clustered in the same clade as that of duck compared to other mammals. Furthermore, this study revealed that chNup210 is expressed in TLR3 signaling pathway and provides fundamental information on Nup210 expression in chicken. Future studies that offer insight into the involvement of chNup210 in the chicken innate immune response against viral infection are recommended.

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.

Antibacterial and Antibiofilm Activities of Diospyros malabarica Stem Extract against Streptococcus mutans (Streptococcus mutans에 대한 인도감나무 줄기 추출물의 항균활성 및 생물막 형성 억제 효과)

  • Kim, Hye Soo;Lee, Sang Woo;Sydara, Kongmany;Cho, Soo Jeong
    • Journal of Life Science
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    • v.29 no.1
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    • pp.90-96
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    • 2019
  • The objective of this study was to evaluate the potential of Diospyros malabarica stem extract, a natural materials, in oral health material. With this aim in mind, thin layer chromatography (TLC), TLC-bioautography, high-performance liquid chromatography (HPLC), electrospray ionization-mass spectrometry (ESI-MS), scanning electron microscopy (SEM), and real-time qPCR were performed. The antibacterial activity of D. malabarica stem extract against Streptococcus mutans KCTC3065 was confirmed in an n-hexane fraction with low polarity. The molecular weight of the antibacterial compound was estimated to be 188 by ESI-MS analysis. The inhibitory effects of the extract on biofilm formation and gene expression related to biofilm formation of S. mutans were determined by SEM and real-time PCR analysis. The extract inhibited the formation of S. mutans biofilms at D. malabarica stem extract concentrations of 1 mg/ml, as shown by SEM. The real-time PCR analysis showed that the expression of the gtfC gene, which is associated with biofilm formation, was significantly decreased in a dose-dependent manner. Based on the above results, it can be concluded that D. malabarica stem extracts, a natural materials, can be used in oral health products to suppress the formation of biofilms by inhibiting tooth adhesion of S. mutans, a causative agent of dental caries.

Cell-Based Screen Using Amyloid Mimic β23 Expression Identifies Peucedanocoumarin III as a Novel Inhibitor of α-Synuclein and Huntingtin Aggregates

  • Ham, Sangwoo;Kim, Hyojung;Hwang, Seojin;Kang, Hyunook;Yun, Seung Pil;Kim, Sangjune;Kim, Donghoon;Kwon, Hyun Sook;Lee, Yun-Song;Cho, MyoungLae;Shin, Heung-Mook;Choi, Heejung;Chung, Ka Young;Ko, Han Seok;Lee, Gum Hwa;Lee, Yunjong
    • Molecules and Cells
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    • v.42 no.6
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    • pp.480-494
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    • 2019
  • Aggregates of disease-causing proteins dysregulate cellular functions, thereby causing neuronal cell loss in diverse neurodegenerative diseases. Although many in vitro or in vivo studies of protein aggregate inhibitors have been performed, a therapeutic strategy to control aggregate toxicity has not been earnestly pursued, partly due to the limitations of available aggregate models. In this study, we established a tetracycline (Tet)-inducible nuclear aggregate (${\beta}23$) expression model to screen potential lead compounds inhibiting ${\beta}23$-induced toxicity. High-throughput screening identified several natural compounds as nuclear ${\beta}23$ inhibitors, including peucedanocoumarin III (PCIII). Interestingly, PCIII accelerates disaggregation and proteasomal clearance of both nuclear and cytosolic ${\beta}23$ aggregates and protects SH-SY5Y cells from toxicity induced by ${\beta}23$ expression. Of translational relevance, PCIII disassembled fibrils and enhanced clearance of cytosolic and nuclear protein aggregates in cellular models of huntingtin and ${\alpha}$-synuclein aggregation. Moreover, cellular toxicity was diminished with PCIII treatment for polyglutamine (PolyQ)-huntingtin expression and ${\alpha}$-synuclein expression in conjunction with 6-hydroxydopamine (6-OHDA) treatment. Importantly, PCIII not only inhibited ${\alpha}$-synuclein aggregation but also disaggregated preformed ${\alpha}$-synuclein fibrils in vitro. Taken together, our results suggest that a Tet-Off ${\beta}23$ cell model could serve as a robust platform for screening effective lead compounds inhibiting nuclear or cytosolic protein aggregates. Brain-permeable PCIII or its derivatives could be beneficial for eliminating established protein aggregates.

The effect of heat stress on frame switch splicing of X-box binding protein 1 gene in horse

  • Lee, Hyo Gun;Khummuang, Saichit;Youn, Hyun-Hee;Park, Jeong-Woong;Choi, Jae-Young;Shin, Teak-Soon;Cho, Seong-Keun;Kim, Byeong-Woo;Seo, Jakyeom;Kim, Myunghoo;Park, Tae Sub;Cho, Byung-Wook
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.8
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    • pp.1095-1103
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    • 2019
  • Objective: Among stress responses, the unfolded protein response (UPR) is a well-known mechanism related to endoplasmic reticulum (ER) stress. ER stress is induced by a variety of external and environmental factors such as starvation, ischemia, hypoxia, oxidative stress, and heat stress. Inositol requiring enzyme $1{\alpha}$ ($IRE1{\alpha}$)-X-box protein 1 (XBP1) is the most conserved pathway involved in the UPR and is the main component that mediates $IRE1{\alpha}$ signalling to downstream ER-associated degradation (ERAD)- or UPR-related genes. XBP1 is a transcription factor synthesised via a novel mechanism called 'frame switch splicing', and this process has not yet been studied in the horse XBP1 gene. Therefore, the aim of this study was to confirm the frame switch splicing of horse XBP1 and characterise its dynamics using Thoroughbred muscle cells exposed to heat stress. Methods: Primary horse muscle cells were used to investigate heat stress-induced frame switch splicing of horse XBP1. Frame switch splicing was confirmed by sequencing analysis. XBP1 amino acid sequences and promoter sequences of various species were aligned to confirm the sequence homology and to find conserved cis-acting elements, respectively. The expression of the potential XBP1 downstream genes were analysed by quantitative real-time polymerase chain reaction. Results: We confirmed that splicing of horse XBP1 mRNA was affected by the duration of thermal stress. Twenty-six nucleotides in the mRNA of XBP1 were deleted after heat stress. The protein sequence and the cis-regulatory elements on the promoter of horse XBP1 are highly conserved among the mammals. Induction of putative downstream genes of horse XBP1 was dependent on the duration of heat stress. We confirmed that both the mechanisms of XBP1 frame switch splicing and various binding elements found in downstream gene promoters are highly evolutionarily conserved. Conclusion: The frame switch splicing of horse XBP1 and its dynamics were highly conserved among species. These results facilitate studies of ER-stress in horse.

Integrative analysis of cellular responses of Pseudomonas sp. HK-6 to explosive RDX using its xenA knockout mutant (Pseudomonas sp. HK-6의 xenA 돌연변이체를 이용하여 RDX 폭약에 노출된 세포반응들의 통합적 분석)

  • Lee, Bheong-Uk;Choi, Moon-Seop;Seok, Ji-Won;Oh, Kye-Heon
    • Korean Journal of Microbiology
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    • v.54 no.4
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    • pp.343-353
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    • 2018
  • Our previous research demonstrated the essential role of the xenB gene in stress response to RDX by using Pseudomonas sp. HK-6 xenB knockout. We have extended this work to examine the cellular responses and altered proteomic profiles of the HK-6 xenA knockout mutant under RDX stress. The xenA mutant degraded RDX about 2-fold more slowly and its growth and survival rates were several-fold lower than the wild-type HK-6 strain. SEM revealed more severe morphological damages on the surface of the xenA mutant cells under RDX stress. The wild-type cells expressed proportionally-increased two stress shock proteins, DnaK and GroEL from the initial incubation time point or the relatively low RDX concentrations, but slightly less expressed at prolonged incubation period or higher RDX. However the xenA mutant did not produced DnaK and GroEL as RDX concentrations were gradually increased. The wild-type cells well maintained transcription levels of dnaA and groEL under increased RDX stress while those in the xenA mutant were decreased and eventually disappeared. The altered proteome profiles of xenA mutant cells under RDX stress also observed so that the 27 down-regulated plus the 3 up-regulated expression proteins were detected in 2-DE PAGE. These all results indicated that the intact xenA gene is necessary for maintaining cell integrity under the xenobiotic stress as well as performing an efficient RDX degradation process.

Evolution of Exercise Performance and Medical Assessments in U-12 Youth Football Players (U-12 유소년 축구선수의 운동수행력 및 의학적 평가)

  • Seo, Sang-Won;Lee, Ho-Seong
    • 한국체육학회지인문사회과학편
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    • v.56 no.5
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    • pp.665-677
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    • 2017
  • This study aimed to examine the evolution of exercise performance and medical assessments in U-12 youth football players. Participants were recruited from an youth football players (YFG, n=15; $11.2{\pm}0.9yrs$) and normal youth (CON, n=15; $11.5{\pm}0.6yrs$). Both groups were tested for exercise performance (health related physical fitness, skill related physical fitness, functional movement screen; FMS and Y-balance) and medical assessments (lower extremity alignment, range of motion; ROM and manual muscle test; MMT). YFG showed a positive effects on pacer in health related physical fitness, 50-m run and Single-leg standing with eyes closed in skill related physical fitness, active straight leg raise and rotary stability in FMS, non-dominant composite score in Y-balance, respectively compared to CON. However, YFG showed a negative effects on Q-angle of dominant and non-dominant in lower extremity alignment, dorsiflexion of non-dominant, plantarflexion of dominant and non-dominan in ROM, hip extension and abduction, knee flexion and extension of dominant, knee flexion and extension of non-dominant in MMT, respectively compared to CON. These results indicated that U-12 youth football players have shown excellent exercise performance, but medical assessments has proved negative effects.

Effects of Overall Shading and Partial Shading on the Response of Chlorophyll Fluorescence of Soybean (전면적차광과 부분차광이 콩 엽록소 형광 반응에 미치는 영향)

  • Cho, Yuna;Jo, Euni;Jeong, Jae-Hyeok;Yoon, Changyong;An, Kyunam;Cho, Jaeil
    • Korean Journal of Agricultural and Forest Meteorology
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    • v.23 no.3
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    • pp.163-168
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    • 2021
  • The growth experiment under shading condition has been performed to understand the eco-physiological responses of crops to light in terms of photosynthesis. There are two types of shading: overall shading and partial shading. In this study, the chlorophyll fluorescence of soybean was observed under the overall shading of the box made by polyresin and the partial shading at agrivoltaic system. The overall shading condition during vegetative growth induced lower SPAD and Electron transport rate (ETR). These lower values recovered after removal of shading box. However, the Non-photochemical fluorescence quenching (NPQ) became lower under overall shading and higher under partial shading. Such increase in NPQ limited crop photosynthesis even though the ETR was almost same to the control without shading treatment. Under the condition of partial shading, the values of SP AD and ETR for soybean did not change. However, the NPQ was higher than control condition. This suggests that the crop photosynthesis under both types of shading would be decreased by different eco-physiological processes which are the lower ETR in overall shading and the higher NP Q in partial shading despite the reduced light under shading conditions.

A Comparative Study on Direct Instrument Methods in Open Channel for Measuring River Water Usage (하천수 사용량 계측을 위한 개수로에서의 직접 계측방법 비교 연구)

  • Baek, Jongseok;Kim, Chiyoung;Lee, Kisung;Kang, Hyunwoong;Song, Jaehyun
    • Journal of Korean Society of Disaster and Security
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    • v.13 no.4
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    • pp.65-74
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    • 2020
  • Continuous and accurate instrument of river water usage is needed for sustainable river water management. Although the instrument methods applicable to each point of use of river water are different, more precise direct instrument methods are required at the point of major open channel. Users of river water should select appropriate direct instrument methods to measure usage, but there is a lack of standards and verification research. In this study, the H-Q rating curve method, ultrasonic method, and microwave method were applied directly to the test basin in the upper basin of Mangyeong river, and the accuracy of measurement data was evaluated by comparing absolute error between discharge data calculated by instrument method. When comparing the calculated discharge of point units, the ultrasonic method showed the best results of the actual measurement. Through continuous instrument, the sum of the daily and monthly units was compared, and the ultrasonic and microwave methods were shown to be highly accurate. Based on the results of this study, it is hoped that the appropriate direct measurement method can be selected according to the importance of the river water use facility, considering that the ultrasonic method and the microwave method are relatively costly compared to the water level-flow relationship method.

Biological and molecular characterization of feline caliciviruses isolated from cats in South Korea

  • Yang, Dong-Kun;Park, Yu-Ri;Yoo, Jae Young;Choi, Sung-Suk;Park, Yeseul;An, Sungjun;Park, Jungwon;Kim, Heui-Jin;Kim, Jongho;Kim, Ha-Hyun;Hyun, Bang-Hun
    • Korean Journal of Veterinary Research
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    • v.60 no.4
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    • pp.195-202
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    • 2020
  • Feline calicivirus (FCV) infection results in a common upper respiratory disease associated with oral ulceration in cats. Although FCV infection has been reported in cats worldwide, the biologic and genetic features of South Korean FCV are unclear. We aimed to investigate the biological and genetic features of South Korean FCV isolates. Crandell-Rees feline kidney (CRFK) cells were used to isolate FCV from 58 organ homogenate samples. The FCV isolates were confirmed by cytopathic effects, immunofluorescence, electron microscopy, and reverse transcription polymerase chain reaction assays. Viral genetic analysis was carried out with VP2 gene and complete genomes of FCVs. Five viruses propagated in CRFK cells were confirmed to be FCVs. The FCV17D283 isolate showed the highest viral titer of 107.2 TCID50/mL at 36 h post-inoculation. Korean FCV isolates did not grow well in Vero, BHK-21, A72, or Madin-Darby canine kidney cells. The FCV17D03 and FCV17D283 isolates had the highest genetic similarity (80.1% and 86.9%) with the UTCVM-H1 and 14Q315 strains, which were isolated in the United States and South Korea in 1995 and 2014, respectively. We isolated five FCVs from cats and detected important genetic differences among them. FCV isolates did not show any virulent effects in mice.