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Characterization of typical Aeromonas salmonicida isolated from Sea-Chum Salmon (Oncorhynchus keta) (해수에 순치된 첨연어(Oncorhynchus keta)에서 분리된 정형 에로모나스 살모니시다(Aeromonas salmonicida)에 대한 특성 분석)

  • Jongwon Lim;Sungjae Ko;Youngjun Park;Do-il Ahn;Suhee Hong
    • Journal of fish pathology
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    • v.36 no.2
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    • pp.263-275
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    • 2023
  • Chum salmon (Oncorhynchus keta) is a species which returns to Korea for spawning and was produced as seed production at the Fisheries Resources Agency located in Uljin-gun, Gyeongsangbuk-do to preserve the species. However, farmed chum salmon showed symptoms of bacterial infection. Therefore, in this study, bacteria were isolated to identify the causative agent from chum salmon in October 2021. The isolated bacteria were identified based on the sequences of 16S rDNA, rpoD (RNA polymerase sigma factor σ70), and vapA (A-layer) genes. Also, salinity-growth curve, biochemical characterization, antibiotic susceptibility test, and pathogenicity analysis were performed in four strains. As a result, four isolated strains were identified as Aeromonas salmonicida subsp. salmonicida. Additionally, the bacterial strains showed a decrease in growth as the salt concentration increased in the medium. All of the isolated strains exhibited γ-hemolysis, and the same biochemical properties. In the antimicrobial susceptibility test, all strains showed an inhibition zone of 40 to 44 mm for oxolinic acid, flumequine, and florfenicol. Pathogenic factors were assessed by RT-PCR at the mRNA level, and found that the four strains expresses the outer membrane ring of T3SS (ascV), inner membrane ring of T3SS (ascC), vapA, enterotoxin (act), and lipase (lip) genes which are well known to significantly contribute to the pathogenicity of A. salmonicida. The results of this study can be used as basic data to prevent A. salmonicida subsp. salmonicida occurring in sea-chum salmon in the future.

Characterization and Evaluation of Melanocortin 4 Receptor (MC4R) Gene Effect on Pork Quality Traits in Pigs (돼지 Melanocortin 4 Receptor (MC4R) 유전자의 육질연관성 분석)

  • Roh, Jung-Gun;Kim, Sang-Wook;Choi, Jung-Suk;Choi, Yang-Il;Kim, Jong-Joo;Choi, Bong-Hwan;Kim, Tae-Hun;Kim, Kwan-Suk
    • Journal of Animal Science and Technology
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    • v.54 no.1
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    • pp.1-8
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    • 2012
  • This study aimed to investigate the single nucleotide polymorphisms (SNPs) of the porcine MC4R gene and validate the effect of the MC4R genotype for marker assisted selection (MAS). Six amplicons were produced to analyze the entire base sequences of the porcine MC4R gene and six SNPs were detected (c.-780C>G, c.-135C>T, c.175C>T-Leu59Leu, c.707A>G-Arg236His, c.892A>G-Asp298Asn, and c.*430A>T). Linkage disequilibrium (LD) of the six SNPs was analyzed by performing haploid analysis. There was a perfect linkage disequilibrium in c.-780C>G, c.-135C>T, c.175C>T-Leu59Leu, c.707A>G-Arg236His, and c.*430A>T. Only the c.892A>G (Asp298Asn) SNP showed a very low LD with an $r^2$ value of 0.028 and the D' value of 0.348. As a result, the two SNPs-c.707A>G (Arg236His) and c.892A>G (Asp298Asn)-were selected to extract the genotype frequencies from the 5 pig breeds by using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) genotype analysis method. The SNP frequency of c.707A>G (Arg236His) indicated the presence of the A (His) allele only in Yorkshire, while the G allele was fixed in the KNP, Landrace, Berkshire, and Duroc. Association analysis was carried out in 484 pigs with the c.707A>G (Arg236His) SNP and the meat quality traits of four different pig cross populations: a significant association was noted in crude fat, sirloin moisture, meat color, and the degree of red and yellow coloration. The frequency of the c.892A>G(Asp298Asn) SNP genotype varied among the breeds; while Duroc showed the highest frequency of the A (Asn) allele, KNP showed the highest frequency of the G (Asp) allele. Association analysis was carried out in 1126 pigs with the c.892A>G (Asp298Asn) SNP and the meat quality traits of four pig populations: a highly significant linkage was noted in the back-fat thickness (P<0.002). It was found that the back-fat thickness was higher in individuals with the AA genotype than in those with the AG or GG genotype. Thus, in this study, we verified that the c.892A>G (Asp298Asn) SNP in the pig MC4R gene has a sufficient effect as a gene marker for MAS in Korean pork industry.

Restoration of endangered orchid species, Dendrobium moniliforme (L.) Sw. (Orchidaceae) in Korea (멸종위기 난과 식물 석곡의 복원)

  • Kim, Young-kee;Kang, Kyung-Won;Kim, Ki-Joong
    • Korean Journal of Plant Taxonomy
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    • v.46 no.2
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    • pp.256-266
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    • 2016
  • A total of 13,000 individuals of Dendrobium moniliforme (L.) Sw. artificially propagated in laboratories and greenhouses were restored in their natural habitat of Bogildo Island, Wandogun, in the southern part of Korea in June of 2013. The growing conditions of the individuals were monitored for two years. The parental individuals for the restoration were obtained from a wild population in southern Korea, from which seeds were produced via artificial crossings. These seeds were germinated and cultivated in growing media and two-year-old plants were then grown in greenhouse beds. The genetic diversity among the propagated individuals was confirmed by examining DNA sequences of five regions of the chloroplast genome and the nuclear ITS region. The diversity values were as high as the average values of natural populations. All propagated individuals were transplanted into two different sites on Bogildo by research teams with local residents and national park rangers. After restoration, we counted and measured the surviving individuals, vegetative propagated stems, and growth rates in June of both 2014 and 2015. There was no human interference, and 97% of the individuals survived. The number of propagules increased by 227% in two years. In contrast, the average length of the stems decreased during the period. In addition, different survival and propagation rates were recorded depending on the host plants and the restored sites. The shaded sides of rock cliffs and the bark of Quercus salicina showed the best propagation rates, followed by the bark of Camellia japonica. A few individuals of D. moniliforme successfully flowered, pollinated, and fruited after restoration. Overall, our monitoring data over two years indicate that the restored individuals were well adapted and vigorously propagated at the restored sites. In order to prevent human disturbance of the restored sites, a CCTV monitoring system powered by a solar panel was installed after the restoration. In addition, a human surveillance system is operated by national park rangers with local residents.

Molecular Phylogenetic Study of Nesiohelix samarangae Based on Metallothionein Gene (동양달팽이(Nesiohelix samarangae)의 metallothionein 유전자를 기초로 한 분자계통 분류학적 연구)

  • Lee, Jun-Seo;Min, Byung-Jun;Kang, Se-Won;Lee, Jae-Bong;Baek, Moon-Ki;Hwang, Seung-Young;Kim, So-Hee;Kho, Weong-Gyu;Choi, Sang-Haeng;Chae, Sung-Hwa;Park, Hong-Seog;Han, Yeon-Soo;Lee, Jun-Sang;Jeong, Kye-Heon;Lee, Yong-Seok
    • The Korean Journal of Malacology
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    • v.24 no.1
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    • pp.73-80
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    • 2008
  • Numerous morphological studies on N. samarangae have been well conducted over the last ten years. In this context, we have attemtped to do molecular phylogenetic analysis by using metallothionein (MT) gene from N. samarangae. To this end, we cloned the full length cDNA of MT from cDNA library of N. samarangae. The complete cDNA sequences were obtained from the expressed sequence tag (EST) sequencing project of N. samarangae, The coding region of 195 bp gives an amino acid sequence of 65 residues including methionine. There are 5' (61 bp) and 3' (48 bp) untranslated region at both ends of the Ns-MT cDNA sequence. The combined results from BLAST analyses, multiple sequence alignment and molecular phylogenetic study of Ns-MT cDNA indicate that N. samarangae has similarity to land snails such as Helix pomatia, Helix aspersa and Arianta arbustorum.

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Expression Pattern of Skeletal-Muscle Protein Genes and Cloning of Parvalbumin mRNA in Dark-banded Rockfish (Sebastes inermis) (볼락(Sebastes inermis) 근육단백질 유전자의 성장단계별 발현 양상과 parvalbumin 유전자 클로닝)

  • Jang, Yo-Soon
    • Korean Journal of Ichthyology
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    • v.23 no.1
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    • pp.1-9
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    • 2011
  • Differentially Expressed Gene (DEG) was obtained from Differential Display Reverse Transcription (DDRT)-PCR using Annealing Control Primer (ACP) to search and clone genes related to developmental stages of Sebastes inermis. By using 120 ACPs, the nucleotide sequences obtained from 16 DEGs showing higher expression in 6-month-old skeletal muscle than 18-month-old ones and from 22 DEGs displaying stronger expression in 18-month-old than 6-month-old were analyzed and BLAST was conducted. The results identified that DEGs shared 69~95% homology with genes of parvalbumin (PVALB), nucleoside diphosphate kinase (NDK) B, tropomyosin (TPM), troponin I (TnI), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), muscle-type creatine kinase (CKM2), small EDRK-rich factor 2 (SERF2), adenosine monophosphate deaminase (AMPD), Trimeric intracellular cation channel type A (TRICA), Rho GTPase-activating protein 15 (ARHGAP15), S-formylglutathione hydrolase (Esterase D; ESD), heat shock protein 70 (hsp70), type 1 collagen alpha 2 (COL1A2), glutathione S-transferase, Mid1-interacting protein 1 (Mid1lip1), myosin light chain 1 (MYL1), sarcoplasmic/endoplasmic reticulum calcium ATPase 1B (SERCA1B), and ferritin heavy subunit (FTH1). Expression pattern by developmental stage of DEG14 and PVALB exhibiting strong expression in 6-month-old skeletal muscle was investigated using real time PCR. Expression was reduced as Sebastes inermis grew. Expression of PVALB gene was extremely low after 6 months of age. Expression of CKM2 showed higher expression in 18-month-old skeletal muscle than in 6-month-old muscles, and increased continuously until 4 years old, after which CKM2 expression became gradually reduced. By analysis of tissue-specific expression patterns of DEG, DEG14 was expressed mainly in skeletal muscle, liver, kidney and spleen tissues, whereas PVALB expression was expressed in skeletal muscle and kidney, but not in liver and spleen tissues. CKM2 was expressed in skeletal muscle, kidney, and spleen tissues, but not in liver tissues. PVALB gene was composed of 110 amino acids, which constituted 659 bp nucleotides. The results reported here demonstrate that the expression patterns of parvalbumin and CKM2 could be used as molecular markers for selecting fishes exhibiting fast growth.

Phylogenetic Analysis based on Metallothionein Gene Sequence of an Indigenous Species Pisidium (Neopisidium) coreanum in Korea (한국 고유종 Pisidium (Neopisidium) coreanum (산골조개) 의 metallothionein 유전자를 기초로 한 분자계통 분류학적 연구)

  • Baek, Moon-Ki;Lee, Jun-Seo;Kang, Se-Won;Lee, Jae-Bong;Kang, Hyun-Jung;Jo, Yong-Hun;Noh, Mi-Young;Han, Yeon-Soo;Choi, Sang-Haeng;Chae, Sung-Hwa;Park, Hong-Seog;Lee, Jun-Sang;Lee, Yong-Seok
    • The Korean Journal of Malacology
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    • v.25 no.2
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    • pp.153-160
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    • 2009
  • Pisidium (Neopisidium) coreanum is a freshwater snail and lives in spring water near mountain areas. Interestingly, this snail has been traditionally regarded as medicinal food, and thus has been used as folk remedies for healing broken bones. Recently, alpha classification on Pisidium (Neopisidium) coreanum through redescription has been conducted. However, not much attention has been made in beta classification. In this study, we performed the beta classification based on metallothionein (MT) genes found from various organisms. To this end, the complete cDNA sequences were obtained from the Expressed Sequence Tag (EST) sequencing project of Pisidium (Neopisidium) coreanum. The coding region (315 bp) encoded an amino acid sequence of 105 residues. The combined results from BLAST analyses, multiple sequence alignment and molecular phylogenetic study of Pc-MT gene indicate that Pisidium (Neopisidium) coreanum has similarity to freshwater bivalve such as Dreissena polymorpha (zebra mussel), Unio tumidus (swollen river mussel) and Crassostrea ariakensis (suminoe oyster).

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Expression of CsRCI2s by NaCl stress reduces water and sodium ion permeation through CsPIP2;1 in Camelina sativa L.

  • Kim, Hyun-Sung;Lim, Hyun-Gyu;Ahn, Sung-Ju
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.194-194
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    • 2017
  • Camelina (Camelina sativa L.) is a potential bio-energy crop that has short life cycle about 90 days and contains high amount of unsaturated fatty acid which is adequate to bio-diesel production. Enhancing environmental stress tolerance is a main issue to increase not only crop productivity but also big mass production. CsRCI2s (Rare Cold Inducible 2) are cold and salt stress related protein that localized at plasma membrane (PM) and assume to be membrane potential regulation factor. These proteins can be divide into C-terminal tail (CsRCI2D/E/F/G) or no-tail group (CsRCI2A/B/C/H). However, function of CsRCI2s are less understood. In this study, physiological responses and functional characterization of CsRCI2s of Camelina under salt stress were analyzed. Full-length CsRCI2s (A/B/E/F) and CsPIP2;1 sequences were confirmed from Camelina genome browser. Physiological investigations were carried out using one- or four-week-old Camelina under NaCl stress with dose and time dependent manner. Transcriptional changes of CsRCI2A/B/E/F and CsPIP2;1 were determined using qRT-PCR in one-week-old Camelina seedlings treated with NaCl. Translational changes of CsRCI2E and CsPIP2;1 were confirmed with western-blot using the antibodies. Water transport activity and membrane potential measurement were observed by cRNA injected Xenopus laevis oocyte. As results, root growth rate and physiological parameters such as stomatal conductance, chlorophyll fluorescence, and electrolyte leakage showed significant inhibition in 100 and 150 mM NaCl. Transcriptional level of CsPIP2;1 did not changed but CsRCI2s were significantly increased by NaCl concentration, however, no-tail type CsRCI2A and CsRCI2B increased earlier than tail type CsRCI2E and CsRCI2F. Translational changes of CsPIP2;1 was constitutively maintained under NaCl stress. But, accumulation of CsRCI2E significantly increased by NaCl stress. CsPIP2;1 and CsRCI2A/B/E/F co-expressed Xenopus laevis oocyte showed decreased water transport activity as 61.84, 60.30, 62.91 and 76.51 % at CsRCI2A, CsRCI2B, CsRCI2E and CsRCI2F co-expression when compare with single expression of CsPIP2;1, respectively. Moreover, oocyte membrane potential was significantly hyperpolarized by co-expression of CsRCI2s. However, higher hyperpolarized level was observed in tail-type CsRCI2E and CsRCI2F than others, especially, CsRCI2E showed highest level. It means transport of $Na^+$ ion into cell is negatively regulated by expression of CsRCI2s, and, function of C-terminal tail is might be related with $Na^+$ ion influx. In conclusion, accumulation of NaCl-induced CsRCI2 proteins are related with $Na^+$ ion exclusion and prevent water loss by CsPIP2;1 under NaCl stress.

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Dual Codec Based Joint Bit Rate Control Scheme for Terrestrial Stereoscopic 3DTV Broadcast (지상파 스테레오스코픽 3DTV 방송을 위한 이종 부호화기 기반 합동 비트율 제어 연구)

  • Chang, Yong-Jun;Kim, Mun-Churl
    • Journal of Broadcast Engineering
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    • v.16 no.2
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    • pp.216-225
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    • 2011
  • Following the proliferation of three-dimensional video contents and displays, many terrestrial broadcasting companies have been preparing for stereoscopic 3DTV service. In terrestrial stereoscopic broadcast, it is a difficult task to code and transmit two video sequences while sustaining as high quality as 2DTV broadcast due to the limited bandwidth defined by the existing digital TV standards such as ATSC. Thus, a terrestrial 3DTV broadcasting with a heterogeneous video codec system, where the left image and right images are based on MPEG-2 and H.264/AVC, respectively, is considered in order to achieve both high quality broadcasting service and compatibility for the existing 2DTV viewers. Without significant change in the current terrestrial broadcasting systems, we propose a joint rate control scheme for stereoscopic 3DTV service based on the heterogeneous dual codec systems. The proposed joint rate control scheme applies to the MPEG-2 encoder a quadratic rate-quantization model which is adopted in the H.264/AVC. Then the controller is designed for the sum of the left and right bitstreams to meet the bandwidth requirement of broadcasting standards while the sum of image distortions is minimized by adjusting quantization parameter obtained from the proposed optimization scheme. Besides, we consider a condition on maintaining quality difference between the left and right images around a desired level in the optimization in order to mitigate negative effects on human visual system. Experimental results demonstrate that the proposed bit rate control scheme outperforms the rate control method where each video coding standard uses its own bit rate control algorithm independently in terms of the increase in PSNR by 2.02%, the decrease in the average absolute quality difference by 77.6% and the reduction in the variance of the quality difference by 74.38%.

Assessment of the Hydration Effect on In Vitro Human Skin by $^1H$ MRS at 14.1T System (고자장 14.1 T MRI/MRS 시스템에서 양성자 자기공명분광법을 이용한 생체 외 인체피부 보습효과에 대한 연구)

  • Choi Chi-Bong;Hong Sung-Tak;Woo Dong-Chul;Yoon Seong-Ik;Yoon Moon-Hyun;Cho Ji-Hyun;Lee Chul-Hyun;Cheong Chae-Joon;Park Sang-Yong;Oh Chil-Hwan;Choe Bo-Young
    • Progress in Medical Physics
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    • v.17 no.2
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    • pp.83-88
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    • 2006
  • Purpose: We Investigated to achieve high resolution magnetic resonance (MR) Imaging and spectra of human skin in vitro with using a 14.1 T MRI/MRS system, and to evaluate the hydration effect of a moisturizer by measuring the skin's water concentration. Materials and Methods: We used the Brukrer 14.1 T MRI/MRS system with a vertical standard bore that was equipped with a DMX spectrometer gradient system (200 G/cm at a maximum 40 A), RF resonators (2, 5 and 10 mm) and Para Vision software. Spin echo and fast spin echo pulse sequences were employed for obtaining the high resolution MR images. The 3D-localized point resolved spectroscopy (PRESS) method was used to acquire the MR spectra. Results: The high resolution MR images and spectra of human skin in vitro were successfully obtained on a 14.IT system. The water concentration of human skin after applying a moisturizer was higher than that before applying a moisturizer. Conclusions: The present study demonstrated that the high-resolution MR images and spectra of human skin from a high field MRS instrument could be applicable to evaluating the hydration state of the stratum corneum.

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Combined Effects of Gamma-irradiation and Hyperthermia on the Human Cell Lines for Various Temperatures and Time Sequences (감마선과 온열치료 병용시 세포 치사 능력 증강에 관한 실험적 연구)

  • Koh Kyung Hwan;Cho Chul Koo;Park Woo Yoon;Yoo Seong Yul;Yun Hyong Geun;Shim Jae Won;Lee Mi Jung
    • Radiation Oncology Journal
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    • v.11 no.1
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    • pp.51-58
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    • 1993
  • We tried to establish the theoretical basis of clinical use of combined modality of hyperthermia and radiation therapy. For this purpose, we made an in vitro experiment in order to get the synergistic and/or additive effects on the cell killing of hyperthermia combined with radiation therapy by using the microwave-hyperthermia machine already installed at our department. In our experiment, we use two human cell lines: MKN-45 (adenocarcinoma of stomach) and K-562 (leukemia cell lines). In cases of combined treatments of hyperthermia and gamma-irradiation, the therapeutic effect was the highest in the simultaneous trial. Hyperthermia after gamma irradiation showed slightly higher therapeutic effect than that before irradiation without significant difference, but its effect was the same in the interval of 6 hours between hyperthermia and irradiation. The higher temperature and the longer treatment time were applied, the higher therapeutic effects were observed. We could observe the thermoresistance by time elapse at $43^{\circ}C$. When hyperthermia was done for 30 minutes at the same temperature, thermal enhancement ratio (TER) at DO. 01 (dose required surviving fraction of 0.01) were $2.5{\pm}0.08,\;3.75{\pm}0.18$, and $5.0{\pm}0.15\;at\;436{\circ}C,\;44^{\circ}C,\;and\;45^{\circ}C$ respectively in K-562 leukemia cell lines. Our experimental data showed that more cell killing effect can be obtained in the leukemia cell lines, although they usually are known to be radiosensitive, when treated with combined hyperthermia and radiation therapy. Furthermore, our data show that leukemia cell lines may have various intrinsic radiosensitivity, especially in vitro experiments. The magnitude of cell killing effect, however, will be less than that of MKN-45.

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