• Title/Summary/Keyword: Species detection

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Feasibility of simultaneous measurement of cytosolic calcium and hydrogen peroxide in vascular smooth muscle cells

  • Chang, Kyung-Hwa;Park, Jung-Min;Lee, Moo-Yeol
    • BMB Reports
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    • v.46 no.12
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    • pp.600-605
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    • 2013
  • Interplay between calcium ions ($Ca^{2+}$) and reactive oxygen species (ROS) delicately controls diverse pathophysiological functions of vascular smooth muscle cells (VSMCs). However, details of the $Ca^{2+}$ and ROS signaling network have been hindered by the absence of a method for dual measurement of $Ca^{2+}$ and ROS. Here, a real-time monitoring system for $Ca^{2+}$ and ROS was established using a genetically encoded hydrogen peroxide indicator, HyPer, and a ratiometric $Ca^{2+}$ indicator, fura-2. For the simultaneous detection of fura-2 and HyPer signals, 540 nm emission filter and 500 nm~ dichroic beamsplitter were combined with conventional exciters. The wide excitation spectrum of HyPer resulted in marginal cross-contamination with fura-2 signal. However, physiological $Ca^{2+}$ transient and hydrogen peroxide were practically measurable in HyPer-expressing, fura-2-loaded VSMCs. Indeed, distinct $Ca^{2+}$ and ROS signals could be successfully detected in serotonin-stimulated VSMCs. The system established in this study is applicable to studies of crosstalk between $Ca^{2+}$ and ROS.

Cryptosporidium hominis Infection Diagnosed by Real-Time PCR-RFLP

  • Cheun, Hyeng-Il;Kim, Kyungjin;Yoon, Sejoung;Lee, Won-Ja;Park, Woo-Yoon;Sim, Seobo;Yu, Jae-Ran
    • Parasites, Hosts and Diseases
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    • v.51 no.3
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    • pp.353-355
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    • 2013
  • There are approximately 20 known species of the genus Cryptosporidium, and among these, 8 infect immunocompetent or immunocompromised humans. C. hominis and C. parvum most commonly infect humans. Differentiating between them is important for evaluating potential sources of infection. We report here the development of a simple and accurate real-time PCR-based restriction fragment length polymorphism (RFLP) method to distinguish between C. parvum and C. hominis. Using the CP2 gene as the target, we found that both Cryptosporidium species yielded 224 bp products. In the subsequent RFLP method using TaqI, 2 bands (99 and 125 bp) specific to C. hominis were detected. Using this method, we detected C. hominis infection in 1 of 21 patients with diarrhea, suggesting that this method could facilitate the detection of C. hominis infections.

Determination of Eleutherosides B and E in Various Parts of Acanthopanax Species (오갈피나무의 부위별 Eleutheroside B와 Eleutheroside E의 정량)

  • Lee, Sang-Hyun;Kang, Sam-Sik;Cho, Seon-Haeng;Ryu, Su-Noh;Lee, Burm-Jong
    • Korean Journal of Pharmacognosy
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    • v.36 no.2 s.141
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    • pp.70-74
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    • 2005
  • High performance liquid chromatography(HPLC) was used for the determination of eleutherosides B and E in various parts of Acanthopanax species. Eleutheroside B was detected in the stem (1.45 mg/g) and the root (0.59 mg/g) of A. senticosus and the stem (0.31 mg/g) of A. senticosus grafted on A. sessiliflorus. But there is no detection of eleutheroside B in A. sessiliflorus and the root of A. senticosus grafted on A. sessiliflorus. Eleutheroside E was detected in the stem (0.92, 0.97 and 0.30 mg/g, respectively) and the root (0.65, 0.94, 0.34 mg/g, respectively) of A. senticosus, A. sessiliflorus and A. senticosus grafted on A. sessiliflorus.

Neofusicoccum ribis Associated with Leaf Blight on Rubber (Hevea brasiliensis) in Peninsular Malaysia

  • Nyaka Ngobisa, A.I.C.;Zainal Abidin, M.A.;Wong, M.Y.;Wan Noordin, M.W.D.
    • The Plant Pathology Journal
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    • v.29 no.1
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    • pp.10-16
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    • 2013
  • Hevea brasiliensis is a natural source of rubber and an important plantation tree species in Malaysia. Leaf blight disease caused by Fusicoccum substantially reduces the growth and performance of H. brasiliensis. The aim of this study was to use a combination of both morphological characteristics and molecular data to clarify the taxonomic position of the fungus associated with leaf blight disease. Fusicoccum species were isolated from infected leaves collected from plantations at 3 widely separated locations - Selangor, Perak, and Johor states - in Peninsular Malaysia in 2010. All the isolates were identified according to their conidial patterns and DNA sequences generated from internal transcribed spacers (ITS1 and ITS2), the 5.8S rRNA, and an unknown locus (BotF15) containing microsatellite repeats. Based on taxonomic and sequence data, Neofusicoccum ribis was identified as the main cause of leaf blight disease in H. brasiliensis in commercial plantations in Malaysia. A pathogenicity trial on detached leaves further confirmed that N. ribis causes leaf blight disease. N. ribis is an important leaf pathogen, and its detection in Malaysia has important implications for future planting of H. brasiliensis.

Bacteriological detection of Brucella abortus and its characterization by PCR in the sporadic outbreak of bovine brucellosis in Gyeonggi province

  • Yang, Su-Jeong;Shim, Hang-Sub;Woo, Jong-Tae;Kim, Hye-Sung;Lee, Sung-Sik
    • Korean Journal of Veterinary Service
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    • v.30 no.2
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    • pp.251-258
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    • 2007
  • Bovine brucellosis has occurred for years in Gyeonggi province under the national test and slaughter scheme. The serum agglutination test (SAT) is a diagnostic tool to confirm the disease despite the argument on its specificity. We selected 8 farms where only one or two individuals were diagnosed as brucellosis through SAT at the primary regular herd check and isolated the causative organism and characterized the species by species-specific PCR. The pathogen isolation was successful in 6 farms out of 8 farms by microbiological culture, showing the successful rate of 75%. The isolation rate of the causative organism represents 70% from supra-mammary lymph node and 60% from uterine tissues. They were characterized as Brucella abortus biovar 1 after biotyping by PCR, showing the fragment of 498 bp. Five of 8 farms were diagnosed as brucellosis two to four times more over the intervals of two or three months. Here in this study we briefly showed the correlation of the sporadic outbreak of brucellosis tested by SAT and the isolation of the causative organism. Moreover one or two reactors against brucellosis among considerable size of herd may indicate that SAT failed to detect potentially infected individuals in the incubation stage or chronic phase of the disease.

Review on metallic components released due to the use of electronic cigarettes

  • Mishra, Vinit K.;Kim, Ki-Hyun;Samaddar, Pallabi;Kumar, Sandeep;Aggarwal, M.L.;Chacko, K.M.
    • Environmental Engineering Research
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    • v.22 no.2
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    • pp.131-140
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    • 2017
  • The use of electronic cigarettes (ECs) is recognized as a source of many pollutants, just like conventional cigarettes (CCs). The analysis of EC aerosol samples has confirmed the presence of various metallic species. Most of these metals originate from various parts of the cartomizer, e.g., solder joints, wires, and silicate beads. The metal concentration levels in EC samples were shown to be generally two to four orders of magnitude lower than those of CCs. However, the use of ECs can still pose significant human health hazards as consumers are exposed to the toxicity of those metals and many other hazardous pollutants released simultaneously via the vaping of ECs. The review also describes the detection and quantification of various metals in ECs and CCs. This review was carried out to assess the level of metal species released from ECs and to suggest proper guidelines to control consumer exposure.

Detection of Seed Born Fungi from the Imported Seed and Effect of Fungicide for Them (수입종자에서의 병원균분리와 약제처리효과)

  • Um Kyung Ho;Sung Jae Mo;Jo Wan Kee;Yu Ki-Yull
    • Korean journal of applied entomology
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    • v.22 no.4 s.57
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    • pp.307-313
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    • 1983
  • Twenty-five species of fungi were deleted and identified from 27 kinds of the intercepted seeds from 5 countries. Eight species of fungi including Fusariun moniliforme were detected from the imported calabash seed (Lagenaria siceraria). These seeds were the most contaminated among the intercepted reeds tested. Six kinds of seeds including larch(Larix kaemferi) were disinfected with Vitathiram, Benlate-T and Captan. Rhynchosporium was detected from seeds treated with Vitathiram whereas Alternaria spp. were detected from seeds treated with Benlate T or Captan. Six genera of fungi were detected from the untreated seeds. In the case of germination, it seem that seed disinfectants do not have any damage on germination of seed.

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Detection of Mitochondrial Reactive Oxygen Species in Living Rat Trigeminal Caudal Neurons

  • Lee, Hae In;Chun, Sang Woo
    • International Journal of Oral Biology
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    • v.40 no.2
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    • pp.103-109
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    • 2015
  • Growing evidence suggests that mitochondrial reactive oxygen species (ROS) are involved in various pain states. This study was performed to investigate whether ROS-induced changes in neuronal excitability in trigeminal subnucleus caudalis are related to ROS generation in mitochondria. Confocal scanning laser microscopy was used to measure ROS-induced fluorescence intensity in live rat trigeminal caudalis slices. The ROS level increased during the perfusion of malate, a mitochondrial substrate, after loading of 2',7'-dichlorofluorescin diacetate ($H_2DCF-DA$), an indicator of the intracellular ROS; the ROS level recovered to the control condition after washout. When pre-treated with phenyl N-tert-butylnitrone (PBN) and 4-hydroxy-2,2,6,6-tetramethylpiperidene-1-oxyl (TEMPOL), malate-induced increase of ROS level was suppressed. To identify the direct relation between elevated ROS levels and mitochondria, we applied the malate after double-loading of $H_2DCF-DA$ and chloromethyl-X-rosamine (CMXRos; MitoTracker Red), which is a mitochondria-specific fluorescent probe. As a result, increase of both intracellular ROS and mitochondrial ROS were observed simultaneously. This study demonstrated that elevated ROS in trigeminal subnucleus caudalis neuron can be induced through mitochondrial-ROS pathway, primarily by the leakage of ROS from the mitochondrial electron transport chain.

Application of Laser Induced Photoacoustic Spectroscopy in the Investigation of Interaction of Neodymium(III) with Water Soluble Synthetic Polymer

  • Tae Hyung Yoon;Hichung Moon;Seung Min Park;Joong Gill Choi;Paul Joe Chong
    • Bulletin of the Korean Chemical Society
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    • v.14 no.5
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    • pp.574-578
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    • 1993
  • Laser-induced photoacoustic spectroscopy (LIPAS), which utilizes the photothermal effect that results from nonradiative relaxation of excited state molecules, was used in the speciation analysis of the complexes of neodymium(III) and water soluble synthetic polyelectrolyte, poly methacrylic acid (PMAA), in 0.1 M $NaClO_4$ at pH of 6.0. The minimum detection limit of Nd(III) by LIPAS was $5.O{\times}10^{-6}$ M. Experiment was carried out at low concentration ratio of Nd(III) to PMAA to assure that 1 : 1 complexes predominate. The bound and free Nd(III) species were characterized by measuring nonradiative relaxation energy of the excited states $(^2GM{7/2}\;and\;^4G_{5/2})$ to the metastable state $(^4G_{3/2})$. Two species were quantified by deconvolution of the mixed spectrum using their respective reference spectra. The conditional stability constant measured by LIPAS was 5.52 L$mol^{-1}$.

Quantitative Analysis of Flavonoid Glycosides in Sophora japonica and Sophora flavescens by HPLC-DAD

  • Kim, Soo Sung;Park, SeonJu;Kim, Nanyoung;Kim, Seung Hyun
    • Natural Product Sciences
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    • v.27 no.4
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    • pp.284-292
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    • 2021
  • Recently, a phytoestrogenic functional food has been developed using the fruits of Sophora japonica. Phytochemical investigation of fruits of S. japonica led to the isolation of eight flavonoid glycosides using various chromatographic techniques. The isolated compounds were identified as genistin (1), sophoricoside (2), genistein 7,4'-di-O-β-D-glucopyransoide (3), sophorabioside (4), genistein-7-O-β-D-glucopyranoside-4'-O-[(α-L-rhamnopyranosyl)-(1→2)-β-D-glucopyranoside] (5), sophoraflavonoloside (6), nicotiflorin (7) and kaempferol-3-O-α-L-rhamnopyranosyl-(1→6)-β-D-glucopyranosyl-(1→3)-β-D-glucopyranoside (8), respectively, by comparison of their spectroscopic data with those reported in the literature. In addition, a new HPLC-DAD method for simultaneous determination of the isolated compounds was developed to quantitate the contents of flavonoids in S. japonica and S. flavescens. The method was validated in terms of limit of detection, limit of quantitation, specificity, linearity, precision and accuracy. The validated method was successfully applied to determine eight flavonoids in two Sophora species. The contents of eight flavonoids varied according to the parts and species. Particularly, it was found that only the fruits of S. japonica contained sophoricoside, a phytoestrogenic isoflavone.