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Saprolegnia parasitica Isolated from Rainbow Trout in Korea: Characterization, Anti-Saprolegnia Activity and Host Pathogen Interaction in Zebrafish Disease Model

  • Shin, Sangyeop;Kulatunga, D.C.M.;Dananjaya, S.H.S.;Nikapitiya, Chamilani;Lee, Jehee;De Zoysa, Mahanama
    • Mycobiology
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    • v.45 no.4
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    • pp.297-311
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    • 2017
  • Saprolegniasis is one of the most devastating oomycete diseases in freshwater fish which is caused by species in the genus Saprolegnia including Saprolegnia parasitica. In this study, we isolated the strain of S. parasitica from diseased rainbow trout in Korea. Morphological and molecular based identification confirmed that isolated oomycete belongs to the member of S. parasitica, supported by its typical features including cotton-like mycelium, zoospores and phylogenetic analysis with internal transcribed spacer region. Pathogenicity of isolated S. parasitica was developed in embryo, juvenile, and adult zebrafish as a disease model. Host-pathogen interaction in adult zebrafish was investigated at transcriptional level. Upon infection with S. parasitica, pathogen/antigen recognition and signaling (TLR2, TLR4b, TLR5b, NOD1, and major histocompatibility complex class I), pro/anti-inflammatory cytokines (interleukin $[IL]-1{\beta}$, tumor necrosis factor ${\alpha}$, IL-6, IL-8, interferon ${\gamma}$, IL-12, and IL-10), matrix metalloproteinase (MMP9 and MMP13), cell surface molecules ($CD8^+$ and $CD4^+$) and antioxidant enzymes (superoxide dismutase, catalase) related genes were differentially modulated at 3- and 12-hr post infection. As an anti-Saprolegnia agent, plant based lawsone was applied to investigate on the susceptibility of S. parasitica showing the minimum inhibitory concentration and percentage inhibition of radial growth as $200{\mu}g/mL$ and 31.8%, respectively. Moreover, natural lawsone changed the membrane permeability of S. parasitica mycelium and caused irreversible damage and disintegration to the cellular membranes of S. parasitica. Transcriptional responses of the genes of S. parasitica mycelium exposed to lawsone were altered, indicating that lawsone could be a potential anti-S. parasitica agent for controlling S. parasitica infection.

Characteristics of Wound Contraction according to the Shape and Antomical Regions of the Wound in Porcine Model (돼지모델에서 상처의 모양과 부위에 따른 상처수축의 특성)

  • Chu, Ho-Jun;Son, Dae-Gu;Kwon, Sun-Young;Kim, Jun-Hyung;Han, Ki-Hwan
    • Archives of Plastic Surgery
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    • v.38 no.5
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    • pp.576-584
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    • 2011
  • Purpose: The shape and location, the amount of the wound and the characteristics of the remaining tissues are known to influence wound contraction. The previous studies using small animals have not been an appropriate model because the wound healing mechanisms and skin structures are different from those of the human. The purpose of this study is to evaluate wound contraction according to the shape and location of the wound using a $Micropig^{(R)}$. Methods: Four $Micropigs^{(R)}$ (Medikinetics, Pyeongtaek, Korea) that were 10 months of age weighed 25 kg were used. Full thickness skin defects were made by clearing all the tissues above the fascial layer in the shape of square, a regular triangle and a circle of 9 $cm^2$ each on the back around the spine. Eight wounds were created on the back of each pig, 50 mm apart from each other. The randomly chosen wound shapes included 11 squares, 11 regular triangles, and 10 circles. Wound dressing was done every other day with polyurethane foam. The wound size was measured using a Visitrak $Digital^{(R)}$ (Smith & Nephew, Hull, UK) on every other day after surgery from day 2 to day 28. A biopsy was performed on day 3, and 1, 2, 3 and 4 weeks to investigate the degree of acute and chronic inflammation, the number of microvesssel and myofibroblast density using H & E stain and immunohistochemistry. The wound contraction rate was calculated to figure out the differences among each of the shapes and the locations. Results: The ultimate shape of the circle wound was oval, and that of the regular triangle and square were stellate. The maximum contraction rate was obtained on 8 to 10 days for all the shapes, which corresponds with the immunohistochemical finding that myofibroblast increases in the earlier 2 weeks whereas it decreases in the later 2 weeks. Epithelialization was seen in the wound margin on day 7 and afterwards. The final wound contraction rates were highest for the regular triangle shapes; however, there were no statistically significant differences. The wound contraction rates by locations showed statistically significant differences. The wound in the cephalic area presented more contractions than that of the wounds in the caudal area. Conclusion: The location of a wound is more important factor than the wound shape in wound contraction.

Biological Activity of Organic Solvent Fractions from Lythrum salicaria L. (Root) (털부처꽃 뿌리로부터 조제된 분획물의 생리활성)

  • Lee, Seung-Eun;Kim, Geum-Soog;Han, Hee-Sun;Lee, Eun-Suk;Kim, Young-Ok;Lee, Jeong-Hoon;Seong, Nak-Sul;Lee, Sang-Won;Kim, Young-Chul
    • Korean Journal of Medicinal Crop Science
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    • v.18 no.5
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    • pp.323-328
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    • 2010
  • Root extract of Lythrum salicaria reported a hepato-protective effect on $CCl_4$-induced liver toxicity of rat was prepared into fractions such as n-hexane up layer (HA), n-hexane down layer (HB), diethyl ether (E), ethylacetate (EA), n-butanol (B) and water (W). Fractions prepared were tested their activities in vitro and in vivo condition. All of the fractions showed effective antioxidant asctivities on DPPH radical and $CuSO_4$-induced oxidation of human low density lipoprotein and E fraction showed the highest inhibitory effect (98.1% at $50\;{\mu}g/m{\ell}$) on linoleic acid autoxidation at $40^{\circ}C$, which was more effective than $\alpha$-tocopherol (82.4%). Five fractions (H = HA plus HB, E, EA, B, and W, 150 mg/kg/day) were fed into Sprague Dawley, male rats for 4 days, which were intoxicated with intra-peritoneal injection of carbon tetrachloride ($1\;m{\ell}/kg$ in corn oil) at the 4th day and were sacrificed in 24 hrs. Serum tumor necrosis factor-alpha (TNF-$\alpha$), a proinflammatory cytokine, elevated with $CCl_4$-intoxication in negative control group ($83\;pg/m{\ell}$) was significantly decreased in E fraction-supplemented group ($18\;pg/m{\ell}$). Cu, Zn-superoxide dismutase (SOD) activity increased in negative control group (0.12 U/mg protein) was decreased in E fraction (0.07 U/mg protein). From the results, it is suggested that ether fraction from root extract of L. salicaria would be a potent antioxidant candidate for ameliorating liver injury induced by chemical intoxicant.

Study on Growth Optimization of InAs/GaSb Strained-Layer Superlattice Structures by High-Resolution XRD Analysis (고분해능 XRD 분석에 의한 InAs/GaSb 응력초격자 구조의 성장 최적화 연구)

  • Kim, J.O.;Shin, H.W.;Choe, J.W.;Lee, S.J.;Kim, C.S.;Noh, S.K.
    • Journal of the Korean Vacuum Society
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    • v.18 no.4
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    • pp.245-253
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    • 2009
  • For the growth optimization of InAs/GaSb (8/8-ML) strained-layer superlattice (SLS), the structure has been grown under various conditions and modes and characterized by the high-resolution x-ray diffraction (XRD) analysis. In this study, the strain modulation is induced by changing parameters and modes, such as the growth temperature, the ratio of V/III beam-equivalent-pressure (BEP), and the growth interruption (GI), and the strain variation is analyzed by measuring the angle separation of 0th-order satellite peak in XRD patterns. The XRD results reveal that the growth temperature and the V/III(Sb/Ga) ratio are major parameters to change the crystallineity and the strain modulation in SLS structures, respectively. We have observed that the SLS samples with compressive strain prepared in this study are show a transition to tensile strain with decreasing V/III(Sb/Ga) ratio, and the GI process is a sensitive factor giving rise to strain modulation. These results obtained in this study suggest that optimized growth temperature and V/III(Sb/Ga) ratio are $350^{\circ}C$ and 20, respectively, and the appropriate GI time is approximately 3 seconds just before InAs growth that the crystallineity is maximized and the strain relaxation is minimized.

Effect of Moutan Cortex Radicis on Gene Expression Profile of Differentiated PC12 Rat Cells Oxidative-stressed with Hydrogen Peroxide (모단피의 PC12 cell 항산화 효과와 관련 HO, MIF, COMT 유전자 발현에 미치는 영향)

  • Son Mu Song;Rho Sam Woong;Ko Eun Jung;Na Youn Gin;Bae Hyun Su;Hong Moo Chang;Shin Min Kyu
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.17 no.4
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    • pp.905-913
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    • 2003
  • Oriental medicine explains aging as the weakening of Kidney-ai, and Kidney-strengthening herbal medicines such as Yukmijihwang-tang have been studied for anti-aging effects. In Western Medicine, the hypothesis that reactive oxidant species(ROS) contribute to the aging process is generally accepted. It has been reported that Moutan Cortex Radicis extract (MCR) was the most effective constituent of Yukmijihwang-tang in decreasing ROS production in oxidative-stressed cells. The purpose of this study is to confirm the anti-oxidant effect of MCR on PC12 cells, the expression of Heme oxygenase (HO), Macrophage migradon inhibitory factor (MIF), Catechol-O-methyltransferase (COMT) using real time RT PCR. PC12 cells were treated without or with hydrogen peroxide in the presence or absence of MCR using MTS assay. Hydrogen peroxide decreased the viability of PC12 cells by 53% and MCR did not influence that of stressed PC 12 cells irrespective of dose or incubation period. However, MCR showed an inhibitory effect on production of ROS in stressed cells, both dose and incubation time dependently. In particular, 1 ㎎/㎖ of MCR for 24 h culture almost returned to normal level. In the quantiation of anti-aging related gene expression, MCR at 1 ㎎/㎖ increased the expression of HO by 370%, MIF by 180% and COMT by 280% through real time RT PCR. In conclusion, MCR treatment protected PC12 cells from hydrogen peroxide and decreased ROS production and enhanced anti-oxidative gene expression such as HO, COMT and MIF, which suggests that MCR is involved in controlling anti-aging of nerve cells through elimination of cytotoxic stimuli.

Functional Analysis of the Stress-Inducible Soybean Calmodulin Isoform-4 (GmCaM-4) Promoter in Transgenic Tobacco Plants

  • Park, Hyeong Cheol;Kim, Man Lyang;Kang, Yun Hwan;Jeong, Jae Cheol;Cheong, Mi Sun;Choi, Wonkyun;Lee, Sang Yeol;Cho, Moo Je;Kim, Min Chul;Chung, Woo Sik;Yun, Dae-Jin
    • Molecules and Cells
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    • v.27 no.4
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    • pp.475-480
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    • 2009
  • The transcription of soybean (Glycine max) calmodulin isoform-4 (GmCaM-4) is dramatically induced within 0.5 h of exposure to pathogen or NaCl. Core cis-acting elements that regulate the expression of the GmCaM-4 gene in response to pathogen and salt stress were previously identified, between -1,207 and -1,128 bp, and between -858 and -728 bp, in the GmCaM-4 promoter. Here, we characterized the properties of the DNA-binding complexes that form at the two core cis-acting elements of the GmCaM-4 promoter in pathogen-treated nuclear extracts. We generated GUS reporter constructs harboring various deletions of approximately 1.3-kb GmCaM-4 promoter, and analyzed GUS expression in tobacco plants transformed with these constructs. The GUS expression analysis suggested that the two previously identified core regions are involved in inducing GmCaM-4 expression in the heterologous system. Finally, a transient expression assay of Arabidopsis protoplasts showed that the GmCaM-4 promoter produced greater levels of GUS activity than did the CaMV35S promoter after pathogen or NaCl treatments, suggesting that the GmCaM-4 promoter may be useful in the production of conditional gene expression systems.

The Changes of Solid, Nitrogen and Phosphorus Concentrations in Pig Slurry Stored at Various Depth of Slurry Storage Tank (돼지분뇨슬러리의 액비조내 저장깊이별 고형물 및 질소, 인의 함량분포에 관한 연구)

  • Jeong, Kwang-Hwa;Chung, Eui-Soo;Park, Chi-Ho;Kwag, Jung-Hoon;Choi, Dong-Yoon;Yoo, Yong-Hee
    • Journal of Animal Environmental Science
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    • v.12 no.3
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    • pp.161-168
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    • 2006
  • This paper describes the changes of characteristics of pig slurry according to storing depth. Most of the substances containing pollutants, such as were Management of manure and wastewater from animal confinement facilities is a critical factor for pollution control. With proper treatment processing method in both solid and liquid forms, it can be used as a fertilizer and soil conditioner. In Korea, liquid-manure handling system is very popular because its treatment and application is easy and labor saving. In the storage tank treatment, the period of fermentation process and solids-liquid separation averages six months and the supernatant liquid is being used as a fertilizer. In this study, the changes in chemical characteristics of pig slurry at varying depths of the storage tank were investigated. Results showed that the pH value of the fermented pig slurry was > 7, while the major pollutants such as BOD, SS, N and P were highest in the bottom of the tank. Therefore, the above findings proved that varying depths in the storage tank can influence the concentration of pollutants of the fermented pig slurry.

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Analysis of Volatile Compounds in Kimchi-Flavored Steak Sauce (김치를 이용한 스테이크소스의 휘발성 향기성분)

  • Cho, Yong-Bum;Park, Woo-Po;Jung, Eun-Joo;Lee, Mi-Jeong;Lee, Yang-Bong
    • Korean Journal of Food Science and Technology
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    • v.34 no.3
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    • pp.351-355
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    • 2002
  • Volatile compounds of kimchi-flavored steak sauce were isolated using simultaneous steam distillation and solvent extraction methods and identified by matching mass spectrum and retention index (RI) with the reference data. Twenty-three compounds including five aldehydes, four alcohols, four acids, three sulfur-containing compounds, one ketone, and six others were identified, among which the most abundant compound was eugenol. Sulfides such as dimethyl dis- and trisulfides, and acids such as acetic and dodecanoic acids showed strong effects on the kimchi flavor. Aroma extract dilution analysis with three fold dilution factor shown in over the FD value 34 identified 2-butanal, 2-pentylfuran, methyl-2-propenyl disulfide, and 2-furanmethanol. Strong good aroma was detected in the RI range between 1030 and 1357, bad aroma in RI 1561, sweety aroma in RI 2057, clover aroma of eugenol in RI 2122, and rancid flavor in RI 2251.

Effects of Al and Mn on the Growth, Nutrient Status and Gas Exchange Rates of Pinus densiflora Seedlings (소나무 묘목(苗木)의 생장(生長), 영양상태(營養狀態) 및 가스교환속도(交換速度)에 미치는 Al과 Mn의 영향(影響))

  • Lee, Choong Hwa;Jin, Hyun-O;Kim, Young-Kul
    • Journal of Korean Society of Forest Science
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    • v.90 no.1
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    • pp.74-82
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    • 2001
  • The effects of Al and Mn concentration on dry weight growth, nutrient status and gas exchange rates of 2-Year-old Japanese red pine(Pinus densiflora) seedlings grown in a nutrient culture solution were investigated. Al was added as aluminum chloride at 0, 10, 30 or 60ppm, and Mn was added as manganese chloride at 0, 30 or 60ppm to the nutrient culture solution. The pH of the solution was maintained at 4.0 by adding HCl or NaOH solution. The seedlings were transplanted into the nutrient culture solution, then they were grown in a greenhouse for 90 days. The interactive effects of Al and Mn on the dry weight growth of the seedlings were not significant. There were a main effect of Al or Mn on the dry weight growth and element concentrations of the seedlings. The treatment with Al of ${\geq}10ppm$ or that with Mn of 60ppm induced a significant reduction in the dry weight growth, which indicates that the effect of Al is stronger than that of Mn. The chlorophyll content of needles was not affected by Al treatment, but was significantly reduced by treatment with Mn of 60ppm. Furthermore, the treatment with Al of 60ppm or that with Mn of ${\geq}30ppm$ caused a significant reduction in the dark respiration rate of the roots. The net photosynthetic rate of the seedlings reduced with increasing the concentration of Al or Mn in the nutrient culture solution, which suggests that Al or Mn induced reductions in the relative growth rate(RGR) and net assimilation rate(NAR) of the seedlings were mainly due to the decrease of net photosynthesis.

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Effects of 18β-glycyrrhetinic acid on pro-inflammatory cytokines and neuronal apoptosis in the hippocampus of lipopolysaccharide-treated mice (18β-Glycyrrhetinic acid가 lipopolysaccharide에 의한 생쥐 뇌조직의 염증성 사이토카인과 해마신경세포 자연사에 미치는 영향)

  • Lee, Ji-Seung;Kwon, Man-Jae;Kweon, Su-Hyeon;Kim, Jeeho;Moon, Ji-Young;Cho, Yoon-Cheong;Shin, Jung-Won;Lee, Jong-Soo;Sohn, Nak-Won
    • The Korea Journal of Herbology
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    • v.31 no.6
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    • pp.73-81
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    • 2016
  • Objectives : $18{\beta}$-Glycyrrhetinic acid (18betaGA) is an metabolite of glycyrrhizin in Glycyrrhiza (licorice). The present study investigated anti-inflammatory and anti-apoptosis effect of 18betaGA on the brain tissue of lipopolysaccharide (LPS)-treated C57BL/6 mice. Methods : 18betaGA was administered orally with low (30 mg/kg) and high (100 mg/kg) doses for 3 days prior to LPS (3 mg/kg) injection. Pro-inflammatory cytokines mRNA including tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), interleukin (IL)-$1{\beta}$, IL-6, and inflammatory enzyme cyclooxygenase-2 (COX-2) mRNA were measured in the cerebral cortex, hippocampus, and hypothalamus tissue using real-time polymerase chain reaction at 24 h after the LPS injection. Histological changes of Cornu ammonis area 1 (CA1) neurons, Bax, Bcl-2, and caspase-3 expression in the hippocampus was also evaluated by immunohistochemistry and Western blotting method. Results : 18betaGA significantly attenuated the up-regulation of TNF-${\alpha}$, IL-$1{\beta}$, IL-6 mRNA, and COX-2 mRNA expression in the brain tissues induced by the LPS injection. 18betaGA also significantly attenuated the reductions of the thickness of CA1 and the number of CA1 neurons. The up-regulation of Bax protein expression in the hippocampal tissue by the LPS injection was significantly attenuated, while the ratio of Bcl-2/Bax expression was increased by 18betaGA treatment. 18betaGA also significantly attenuated the up-regulation of Bax and caspase-3 expression in the CA1 of the hippocampus. Conclusion : This results indicate that 18betaGA has anti-inflammatory and anti-apoptosis effect under neuroinflammation induced by the LPS injection and suggest that 18betaGA may be a beneficial drug for various brain diseases accompanied with the brain tissue inflammation.