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Multiple shoot induction and callus regeneration in Sarcostemma brevistigma Wight & Arnott, a rare medicinal plant

  • Thomas, T. Dennis;Shankar, Surabhi
    • Plant Biotechnology Reports
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    • v.3 no.1
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    • pp.67-74
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    • 2009
  • An efficient micropropagation protocol based on multiple shoot induction and callus regeneration has been standardized in Sarcostemma brevistigma, a rare medicinal plant. The nodal cuttings were cultured on MS medium supplemented with BA ($0.5-8{\mu}M$) or Kn ($0.5-8{\mu}M$) alone or in combination with NAA ($0.5-1.5{\mu}M$). Maximum multiple shoot induction was observed on MS medium supplemented with $4{\mu}M$ BA. On this medium, 100% cultures responded with an average number of 11.3 shoots per explant. However, the average shoot length was limited to only 0.9 cm on this medium. The addition of $1{\mu}M$ NAA along with $4{\mu}M$ BA gave rise to an average number of 10.9 shoots with an average shoot length of 1.8 cm. Luxuriantly growing callus was obtained on MS medium supplemented with BA ($5{\mu}M$) and 2,4-D ($2{\mu}M$). The callus was subcultured on MS medium supplemented with BA ($2-15{\mu}M$) or Kn ($2-15{\mu}M$) alone or in combination with NAA ($0.5-2{\mu}M$) for shoot organogenesis. Optimum callus regeneration was obtained on MS medium supplemented with $10{\mu}M$ BA and $1{\mu}M$ NAA. On this medium, 100% cultures responded with an average number of 13.4 shoots per culture. The shoots obtained via multiple shoot induction and organogenesis were rooted on half-strength MS medium supplemented with NAA ($1-7{\mu}M$) or IBA ($1-7{\mu}M$). IBA was better than NAA in terms of both the percentage of cultures that responded and the average number of roots per explant. The rooted shoots were successfully transplanted to soil with 86% success. This standardized protocol will help to conserve this rare medicinal plant.

Bioremediation on the Benthic Layer in Polluted Inner Bay by Promotion of Microphytobenthos Growth Using Light Emitting Diode (LED) 1. Effects of irradiance and wavelength on the growth of benthic diatom, Nitzschia sp. (발광다이오드(LED)를 이용한 저서미세조류의 성장촉진에 의한 오염해역 저질환경개선 1. 저서규조류 Nitzschia sp. 성장에 영향을 미치는 광량과 파장)

  • Oh, Seok-Jin;Park, Dal-Soo;Yang, Han-Soeb;Yoon, Yang-Ho;Honjo, Tsuneo
    • Journal of the Korean Society for Marine Environment & Energy
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    • v.10 no.2
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    • pp.93-101
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    • 2007
  • In order for bioremediate the benthic layer in polluted inner Bay, the effects of irradiance and wave-length irradiated from light emission diode (LED) on the growth of benthic diatom Nitzschia sp. (Hakozaki Bay strain of Japan) were investigated. The Nitzschia sp. was cultured under blue LED (450 nm), yellow LED (590 nm), red LED (650 nm) and fluorescent lamp (mixed wavelengths). At $25^{\circ}C$ and 30 psu, the growth of Nitzschia sp. showed its peak at $20\;{\mu}mol\;m^{-2}\;s^{-1}$ (blue LED) and $40\;{\mu}mol\;m^{-2}\;s^{-1}$ (fluorescent lamp), and was inhibited at the irradiance higher than that irradiance. Nitzschia sp. in yellow LED and red LED is fitted by a rectangular hyperbolic curve because no photoinhibition was observed under maximum irradiance used in this study. The irradiance-growth curves were described as ${\mu}=-0.46{\exp}(1-I/6.32)+0.46-0.00043I,\;(r^2=0.98)$ under blue LED, ${\mu}=0.42(I+7.87)/(I+58.9),\;(r^2=0.99)$ under yellow LED, ${\mu}=0.39(I+3.39)/(I+21.6),\;(r^2=0.94)$ under red LED, ${\mu}=-0.38{\exp}(1-I/7.23)+0.38-0.00016I,\;(r^2=0.96)$ under fluorescent lamp. Maximum specific growth rate of blue LED, yellow LED, red LED and fluorescent lamp was $0.44\;day^{-1},\;0.42\;day^{-1},\;0.39\;day^{-1}$ and $0.37\;day^{-1}$, respectively. The absorption coefficient ($a_{ph}$) of Nitzschia sp. was similar under all the wavelengths (400 nm-700 nm), although maximum $a_{ph}$ was $0.0224\;m^2\;mg\;chi.\;{\alpha}^{-1}$ in 472 nm and $0.0179\;m^2\;mg\;chi.\;{\alpha}^{-1}$) in 663 nm. The results may indicate the possibility of environmental improvement around the benthic layer in polluted coastal area because microphytobenthos growth is stimulated by means of irradiated blue LED at the benthic boundary layer during both autumn and winter, and yellow LED, which might have been suppressed growth of harmful algae, at the layer during both spring and summer.

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MMP and TIMP production in periodontal ligament fibroblasts stimulated by Prevotella nigrescens lipopolysaccharide (Prevotella nigrescens lipopolysaccharide로 자극한 치주인대 섬유아세포에서 기질금속단백분해효소와 단백분해효소억제제의 생성 양상에 대한 연구)

  • Yang, Won-Kyung;Lee, Woo-Cheol;Kim, Mi-Ri;Son, Ho-Hyun
    • Restorative Dentistry and Endodontics
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    • v.30 no.5
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    • pp.372-384
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    • 2005
  • The purpose of this study was to monitor the secretion of matrix metalloproteinase (MMP) and tissue inhibitor of metalloproteinase (TIMP) by human periodontal ligament (PDL) fibroblasts stimulated with Prevotella nigrescens lipopolysaccharide (LPS), and to examine the effect of calcium hydroxide treatment on P. nigrescens LPS. LPS was extracted and purified from anaerobically cultured P. nigrescens. PDL fibroblasts were stimulated by the LPS (0, 0.1, 1, 10 ${\mu}g/ml$) or LPS (10 ${\mu}g/ml$) pretreated with 12.5 mg/ml of $Ca(OH)_2$ for 3 days, for various periods of time (12, 24, 48 h). Immunoprecipitation were performed for protein level analysis of MMP-1 MMP-2 and TIMP-1. Total RNA was isolated and real-time quantitative polymerase chain reaction (PCR) was performed for quantification of MMP-1 mRNA. According to this study, the results were as follows: 1. The p개duction of MMP-1 by stimulation with P. nigrescens LPS increased in time-dependent manner, and showed maximum value at 48 h in both protein and mRNA level. But there was no dose-dependent increas. 2. MMP-2 production time-dependently increased when stimulated with 1 and 10 ${\mu}g/ml$LPS, but there was no dose-dependent increase. 3. TIMP-1 p개duction increased to 24 h, but decreased at 48 h. It increased when stimulated with 0.1 and 1${\mu}g/ml$, but suppressed at 10 ${\mu}g/ml$ .4. P. nigrescens LPS pretreated with $Ca(OH)_2$ markedly downregulated MMP-1 gene expression.

Organic Compounds in the Nak Dong River and Its Toxicity (낙동강 수질중 유기물질과 독성)

  • 류병호;심종환;최진택;조현철;정종순
    • Journal of Environmental Health Sciences
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    • v.20 no.1
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    • pp.39-53
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    • 1994
  • This study aims to investigate organic compounds and its toxicity by Ames test and chromosomal aberration in the water of the Nak Dong River. Six sampling sites such as Goryung, Hagueun, Maelie, Duksan, Haedong and Myungiang were selected for these pur15oses. 200 l water samples were absorbed on XAD-2 resin columns (2.5X30cm), eluted with organic solvents mixture of acetone: cyclohexane and then dried under vacuum condition. The extracts from the XAD-2 resin was injected into GC/MS and 184 organic compounds were identified such as aldehydes, aromatic compounds, ketones, phenols, hydrocarbons, alcohols, carboxylic acids, alkanes and some unknowns. The US EPA priority pollutants such as naphthlene, bis(2-ethylhexyl)phthalate and other pollutants, 1,2-diethyl benzene, 1,2,3,4-tetrahydronaphthalene and cyclohexanol were detected in these samples. The concentration of chemical pollutants such as 1,2-diethyl benzene, nephthalene, 1,2,3,4-tetrahydronaphthalene, bis(2-ethylhexyl)phthalate and cyclohexanol were ranged into 1.228 $\mu$g/l, 298 $\mu$g/l, 30.191 $\mu$g/l, 1.147 $\mu$g/l and 2.839 $\mu$g/l, respectively. The mutagenic activity of XAD-2 extracts were tested on Salmonella typhimurium TA 98, TA 100, TA 1535 and TA 1537 and then exhibited strong mutagenic activity against S. typhimurium TA 98 and TA 100 in the presence of S$_9$. Amon them, bis(2-ethylhexyl)phtalate and 1,2-diethyl benzene showed the most strongest mutagenic activity against S. typhimurium TA 98 and TA 100 in the presence of S$_9$. On the other hands, chromosomal aberration of XAD-2 extracts in the human blood cells were not occurred by the sampling water at Goryung, Hagueun, Maelie and Duksan, Chromosomal aberration were also not occurred by the each concentration of 0.05, 0.1 amd 0.3 mg/l of each 1,2-diethyl benzol, bis(2-ethylhexyl)phthalate, naphthalene, phenol, cyclohexanol and benzothiazol test solution.

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Comparison of soil nutrients, pH and electrical conductivity among fish ponds of different ages in Noakhali, Bangladesh

  • Tapader, Md. Morshed Alam;Hasan, Mehedi Mahmudul;Sarker, Bhakta Supratim;Rana, Md. Enayet Ullah;Bhowmik, Shuva
    • Korean Journal of Agricultural Science
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    • v.44 no.1
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    • pp.16-22
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    • 2017
  • An experiment was conducted to detect aquaculture pond bottom soil nutrients, pH and electrical conductivity with a view to optimize production and to incorporate the scientific method of fish nursing, rearing and culturing at Noakhali district in Bangladesh. The soil samples were collected from the recently dug ponds (1 - 5 years) and older ponds (> 5 years). Samples were taken from five different spots in a Z shape from each pond and were mixed to get a composite sample. The composite samples from the ponds were collected in polyethylene bags and shipped to the laboratory for analysis. The soil samples were analyzed with respect to pH, electrical conductivity (EC), organic carbon (OC), organic matter (OM), nitrogen (N), phosphorous (P), potassium (K) and sulfur (S). The average value of pH, OC, OM, N, P, K and S were $7.43{\pm}0.40$, $2.21{\pm}1.43%$, $1.47{\pm}0.53%$, $2.52{\pm}0.94{\mu}g\;g^{-1}$, $0.126{\pm}0.047{\mu}g\;g^{-1}$, $3.84{\pm}1.77{\mu}g\;g^{-1}$, $0.191{\pm}0.106{\mu}g\;g^{-1}$ and $306.72{\pm}222.05{\mu}g\;g^{-1}$ respectively, in Noakhali. The average EC, OC, OM, N and P contents were found to be higher in Subornachar than those in Sonapur. On the other hand pH, K and S were found to be higher in Sonapur than the values of Subornachar. The pH, EC, OC, OM, N and S contents were found to be higher in new ponds than old ponds whereas P and K contents were found to be higher in old pond than in new pond.

Isolation of the Antimicrobial Compounds from Aralia cordata Thunb. Extract (독활 뿌리 추출물에서 항균물질의 분리 및 항균성)

  • Han, Wan-Soo
    • Korean Journal of Medicinal Crop Science
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    • v.13 no.4
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    • pp.182-185
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    • 2005
  • Bioassay-guided isolation of the dried roots of Aralia continentalis led to the isolation of (-)-pimara-8(14), 15dien-19-oic acid (continentalic acid) and $(24E)-Stigmasta-5,22-dien-3{\beta}-ol$ (stagmasterol). Their structures were elucidated using $^1H-NMR$, $^{13}C-NMR$, UV and mass spectra analyses. The gram-positive bacterial, including methicilline-resistant (MRSA), were more sensitive to the continentalic acid and stagmasterol than gram-negative bacterial.

Evaluation of Circle Machining Surface Roughness on the Process Conditions using Neural Network (신경회로망을 이용한 가공조건에 따른 원형가공 표면거칠리 평가)

  • Sung, Baek-Sup;Kim, Ill-Soo;Cha, Yong-Hun
    • Journal of the Korean Society of Safety
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    • v.17 no.1
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    • pp.11-17
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    • 2002
  • The purpose of this study was to present the method to choose the optimization machining condition for the wire electric machine. This was completed by examining the ever-changing quality of the material and by improving the function of the wire electric discharge machine. Precision metal mold products and the unmanned wire electric discharge machining system were used and then applied in industrial fields. This experiment uses the wire electric discharge machine with brass wire electrode of 0.25mm. To measure the precision of the machining surface, average values are obtained from 3 samples of measures of center-line average roughness by using a third dimension gauge and a stylus surface roughness gauge. In this experiment, we changed no-node voltage to 7 and 9, pulse-on-time to $6{\mu}s,\;8{\mu}s$ and $10{\mu}s$, pulse-off-time to $8{\mu}s,\;10{\mu}s$ and $13{\mu}s$, and experimented on wire tension at room temperature by 1000gf, 1200gf, and 1400gf, respectively.

Relationship of Nitrate Reductase Activity to Leaf Yield, Protein, Sugar and Physiological Attributes in Mulberry (Morus alba L.)

  • Ghosh, M.K.;Das, B.K.;Das, C.;Mishra, A.K.;Mukherjee, P.K.;Urs, S.Raje
    • International Journal of Industrial Entomology and Biomaterials
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    • v.8 no.1
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    • pp.67-71
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    • 2004
  • Ten improved mulberry varieties (Vl, C1730, C2016, C2017, Anantha, RFS-175, Thallaghatapura, Vishala, S1 and S1635) were evaluated through enzyme assay and estimation of soluble protein content followed by regression analysis, grown under irrigated conditions in the alluvial soils of Gangetic plains of West Bengal in India for five successive crops in a year, The nitrate reductase (EC No. 1.6.6.1) activity (NRA, $\mu$mol N $O_2$- $h^{-1}$ $g^{-1}$ fr, wt.), total soluble protein (mg $g^{-1}$ fr, wt.) was estimated which showed to vary significantly in the tested varieties. In addition to these, the other parameters like unit leaf fresh and dry weight (g), moisture %, unit leaf area ($\textrm{cm}^2$), specific leaf weight (g c $m^{-2}$ ), total soluble sugar (mg $g^{-1}$ fr, wt.), leaf yield/plant (kg), shoot yield/plant (kg) and net photosynthetic rate (NPR, $\mu$$m^{2}$ $s^{-1}$ ) were also studied which showed to vary significantly in tested varieties. Among them, S1635, haying higher NRA (13.25 $\mu$㏖ N $O_2$- $h^{-l}$ $g^{-1}$ fr, wt.), total soluble protein (39.63mg $g^{-1}$ fr, wt.), NPR(16.66 $\mu$$m^{-2}$ $s^{-1}$ ), total soluble sugar (48.44 mg $g^{-1}$ fr. wt.), leaf yield/plant (0.689 kg) and shoot yield/plant (1.135 kg) showed its superiority over other tested varieties. Regression and correlation coefficients were analysed, and a strong positive correlation was found between NRA & total soluble protein, NRA & NPR, NRA & total soluble sugar, NRA af unit leaf weight, NRA & specific leaf weight, NRA & leaf yield/plant, NRA & shoot yield/plant, NPR & leaf yield and NPR & specific leaf weight.t.

Effect of Harvest Time and Infective Juvenile Size of the Entomopathogenic Nematode, Steinernema arenarium, on Pathogenicity, Development, and Propagation (곤충병원성선충, Steinernema arenarium의 수확시기와 유충 체장이 병원성과 발육 및 증식에 미치는 영향)

  • Han, Gun-Yeong;Lee, Dong-Woon;Choo, Young-Moo;Choo, Ho-Yul
    • Korean journal of applied entomology
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    • v.51 no.1
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    • pp.9-18
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    • 2012
  • The size of infective $Steinernema$ $arenarium$ juveniles is variable and ranges from 724 to 1408 ${\mu}m$. Effects of harvest time and infective juvenile size on pathogenicity, development, and reproduction were examined in the last instar of the great wax moth, $Galleria$ $mellonella$. Harvest time of infective juveniles (IJs) of $S.$ $arenarium$ affected pathogenicity. IJs harvested at the 10th day from trapping were more pathogenic than those harvested the 3rd day from trapping. Mortality of $G$. mellonella also depending on harvest time, $i.e$, 100% died within 48h when IJs were harvested at the 10th day, without relation to size. However, mortality was 40% in the small size group (SSG) compared with 18% in the large size group (LSG) within 48h when IJs were harvested at the 3rd day. Establishment of $S.$ $arenarium$ within the host was different depending on IJ size. The number of established IJs was 1.8 in the SSG, 3.3 in the LSG, and 3.2 in the mixed size group (MSG) when IJs were harvested at the 3rd day, and 5.3 in the SSG, 7.4 in the LSG, and 7.6 in the MSG when IJs were harvested at the 10th day. The length of the female adult was 7,070.5 ${\mu}m$ in the SSG and 7,893.9 ${\mu}m$ in the LSG and that of the male was 1,460.5 ${\mu}m$ in the SSG and 1,688.2 ${\mu}m$ in the LSG when IJs were harvested at the 3rd day. The length of the female adult was 7,573.6 ${\mu}m$ in the SSG and 8,305.4 ${\mu}m$ in the LSG and that of the male adult was 1,733.4 ${\mu}m$ in the SSG and 1,794.4 ${\mu}m$ in the LSG when IJs were harvested at the 10th day. Harvest time and size of IJs did not influence numbers of progeny or size of IJS.

Influence of Humidity Variation on the Surface Deffects and Soft Magnetic Properties in the Fabrication of Fe Based Amorphous Alloy Ribbon by the PFC Process (PFC프로세스 의한 Fe기 Fe78Si9B13 비정질 합금리본 제조에 있어서 습도변화가 표면결함 및 연자기적 특성에 미치는 영향)

  • Choi, Y.J.;Jang, S.J.;Kim, S.W.;Jeon, B.S.;Kim, S.M.;Song, C.B.;Kim, Y.C.;Namkung, J.
    • Journal of the Korean Magnetics Society
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    • v.25 no.1
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    • pp.4-9
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    • 2015
  • This study was carried out to investigate a influence of humidity variation (%) on the magnetic properties and the surface flaws in the fabrication of Fe-based $Fe_{78}Si_9B_{13}$ amorphous alloy ribbon by Planar Flow Casting process. As a result, the size of the air pocket and the droplet which is observed in the contact surface and the free face of the amorphous alloy ribbon becomes large when the humidity increases and the size highly increases with the surface roughness at the same time. Especially, the surface roughness value which is made in the 65 % of the humidity is the lowest in the contact surface ($Ra=0.60{\mu}m$, $Rz=3.11{\mu}m$) and the free face ($Ra=0.47{\mu}m$, $Rz=3.00{\mu}m$). Also, in case of the soft magnetic property of the magnetic core which is made with the toroidal core of $23(OD)^*20(ID)^*20(H)$ size, in the sample of the amorphous alloy ribbon which is made in 65% of the humidity, the most excellent value is gained as $B_s(B_{700})=1.055T$, $H_c=0.083Oe$, permeability = 1,197 and core loss = 0.276W/kg.