• Title/Summary/Keyword: Reagent-based method

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Development of a Rapid Detection Method for Potato virus X by Reverse Transcription Loop-Mediated Isothermal Amplification

  • Jeong, Joojin;Cho, Sang-Yun;Lee, Wang-Hyu;Lee, Kui-jae;Ju, Ho-Jong
    • The Plant Pathology Journal
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    • v.31 no.3
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    • pp.219-225
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    • 2015
  • The primary step for efficient control of viral diseases is the development of simple, rapid, and sensitive virus detection. Reverse transcription loop-mediated isothermal amplification (RT-LAMP) has been used to detect viral RNA molecules because of its simplicity and high sensitivity for a number of viruses. RT-LAMP for the detection of Potato virus X (PVX) was developed and compared with conventional reverse transcription polymerase chain reaction (RT-PCR) to demonstrate its advantages over RT-PCR. RT-LAMP reactions were conducted with or without a set of loop primers since one out of six primers showed PVX specificity. Based on real-time monitoring, RT-LAMP detected PVX around 30 min, compared to 120 min for RT-PCR. By adding a fluorescent reagent during the reaction, the extra step of visualization by gel electrophoresis was not necessary. RT-LAMP was conducted using simple inexpensive instruments and a regular incubator to evaluate whether RNA could be amplified at a constant temperature instead of using an expensive thermal cycler. This study shows the potential of RT-LAMP for the diagnosis of viral diseases and PVX epidemiology because of its simplicity and rapidness compared to RT-PCR.

Application of Polyurethane Foam Loaded with a Schiff Base Ligand for Determination of Trace Amounts of Copper in Water Samples by Flame Atomic Absorption Spectroscopy

  • Moftakhar, Mahdieh Koorehpazan;Yaftian, Mohammad Reza;Nahaei, Samaneh;Zamani, Abbas Ali
    • Journal of the Korean Chemical Society
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    • v.58 no.3
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    • pp.283-288
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    • 2014
  • A column solid phase extraction procedure based on modified polyurethane foam (PUF) by a newly synthesized Schiff base ionophore, named 2,2'-{iminobis[propane-3,1-diylnitrilo(1E)prop-1-yl-1-ylidene]}diphenol, was developed for preconcentration step of trace amounts determination of copper ions in water samples by atomic absorption flame spectroscopy. The influence of parameters on the adsorption process such as sample pH, amount of modified PUF packed in the column, type and volume of stripping reagent and its flow rate were investigated and optimized. Under optimum experimental conditions, the calibration graph was linear in a relatively wide range ($0.005-210{\mu}g/ml$) with a limit of detection $0.002{\mu}g/ml$ of copper. The proposed method allows achieving to a concentration factor of >133. The capacity of a column (1.6 cm i.d.) packed by 6 g of PUF modified by 12 mg of the Schiff base was found to be $247.7({\pm}2.1){\mu}g$ of copper. It was found that the adsorption process was highly selective towards copper ions with respect to some associated metal ions. The presented procedure was successfully applied for determination of copper in some water samples.

Ursolic Acid Activates Intracellular Killing Effect of Macrophages During Mycobacterium tuberculosis Infection

  • Podder, Biswajit;Jang, Woong Sik;Nam, Kung-Woo;Lee, Byung-Eui;Song, Ho-Yeon
    • Journal of Microbiology and Biotechnology
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    • v.25 no.5
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    • pp.738-744
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    • 2015
  • Tuberculosis is one of the most threatening infectious diseases to public health all over the world, for which Mycobacterium tuberculosis (MTB) is the etiological agent of pathogenesis. Ursolic acid (UA) has immunomodulatory function and exhibits antimycobacterial activity. However, the intracellular killing effect of UA has yet to be elucidated. The aim of this study was to evaluate the intracellular killing effect of UA during mycobacterial infection. The intracellular killing activity of UA was evaluated in the macrophage cell line THP-1 by the MGIT 960 system as well as by CFU count. The production of reactive oxygen species (ROS) and the level of nitric oxide (NO) were measured using DCF-DA and Griess reagent, respectively. Phagocytosis was observed by a fluorescence-based staining method, and the colony forming units were enumerated on 7H11 agar medium following infection. In addition, MRP8 mRNA expression was measured by qRT-PCR. UA significantly decreased the number of intracellular Mycobacterium through generation of ROS and NO. In addition, it profoundly activated the phagocytosis process of THP-1 cells during MTB-infection. Furthermore, our data demonstrated that UA activated the phagocytosis process in human monocyte cells through MRP8 induction. These data suggest that UA firmly contributes to the intracellular killing effect of macrophages during mycobacterial infection.

Production of Oleamide, a Functional Lipid, by Streptomyces sp. KK90378

  • Kwon, Jeong-Ho;Hwang, Sung-Eun;Han, Jae-Taek;Kim, Chang-Jin;Rho, Jung-Rae;Shin, Jong-Eon
    • Journal of Microbiology and Biotechnology
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    • v.11 no.6
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    • pp.1018-1023
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    • 2001
  • Oleamide (cis-9-octadecenamide) is endogenous primary amide of fatty acid that is produced in small amounts in animal brains. It is known to induce sleep and to lower temperature by destroying the lipid plasma membrane structure of cells, thereby disclosing gap junction channels. To develop a new biological production method for oleamide, a screening program was conducted to isolate a microorganism producing oleamide. Among 1,500 soil microorganisms tested, KK90378 exhibited a potent positive reaction with Dragendoff`s reagent, used to detect the primary amide of oleamide. KK90378 was identified as a Streptomyces species based on cultural and morpohological characteristics, the presence of diaminopimelic acid in the cell wall, and the sugar patterns for the whole-cell extrat. Streptomyces sp. KK90378 produced oleamide 3 days after culture at $28^{\circ}C$, pH 7.2 A series of purification steps, including hexane extraction, silica gel column, and preparative thin layer chromatographies, were performed for the purification of oleamide. A spectrophotometric analysis using $^1H$, $^13C$-NMR, and GC-MS confirmed that the chemical structure of the purified oleamide was identical to that of authentic oleamide.

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Prevalence of Feline Blood Types in Seoul and Kangwon Area of Korea (서울 및 강원지역 고양이의 혈액형 빈도 조사)

  • Ban, Ji-Min;Shin, Ji-Hye;Kim, Jae-Young;Hyun, Chang-Baig;Kim, Doo;Pak, Son-Il
    • Journal of Veterinary Clinics
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    • v.25 no.4
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    • pp.227-230
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    • 2008
  • To determine the distribution of feline blood types and then to estimate the risk of neonatal isoerythrolysis (NI) in non-pedigree cats, we typed blood of 482 cats of both genders and various breeds (336 domestic shorthair cat and 146 pedigree) from August 2005 through July 2007. Blood samples from Seoul and Kangwon province were typed within 5 days after collection by the simple tube method. High-titer anti-A antiserum and anti-B reagent, prepared with Triticum vulgaris lectin, were used to determine type A and type B blood, respectively. The majority of cats were type A (n = 465, 96.5%) and only 3.5% (n = 17) were type B. No type AB blood were detected. Blood type distributions among the non-pedigree and pedigree cats were similar: for non-pedigree cats, 96.4% were type A and 3.6% were type B, whereas for pedigree cats, 96.6% were type A and 3.4% were type B. All type B cats had a very strong agglutination reaction to anti-A antiserum: 8 sample for 3+ and 9 for 4+. Assuming 19% of estimated frequency for the type-B allele in domestic cats, the calculated proportion of random mating from this population at risk for developing NI was 3.4%. Based on this finding, it is strongly recommended that blood typing be performed prior to any blood transfusion or breeding to minimize blood type incompatibilities. Further comprehensive studies on the titer of naturally occurring antibodies in cat populations in Korea and the prevalence of possible NI in practice are clearly required.

Comparison of Serum Fucose Levels in Leukoplakia and Oral Cancer Patients

  • Rai, Narendra Prakash;Anekar, Jayaprasad;Shivaraja, Shankara YM;Divakar, Darshan Devang;Al Kheraif, Abdulaziz Abdullah;Ramakrishnaiah, Ravikumar;Sebastian, Roopa;Raj, AC;Al-Hazmi, Ali;Mustafa, habil Mohamed
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.17
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    • pp.7497-7500
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    • 2015
  • Background: Tumor markers, designated as a broad group of substances produced by malignancies, could be in the form of biochemical substances, immunological substances, cell surface changes and genetic alterations. Cancer, a disorder of cellular behavior is characterized by alteration of serum glycoproteins. L-fucose, a hexose, which is the terminal sugar in most of the plasma glycoproteins, may be useful as a tumor marker for the detection, monitoring and prognostic assessment of malignancies. The aim of the study was to ascertain the role of serum fucose as a biomarker for early detection of oral cancer and to compare serum fucose levels in healthy controls, leukoplakia and oral cancer patients. Materials and Methods: The study included 60 (100.0%) subjects, who were grouped as 20 (33.3%) control subjects, 20 (33.3%) squamous cell carcinoma patients and 20 (33.3%) leukoplakia patients. Fucose estimation was done using UV-visible spectrophotometry based on the method as adopted by Winzler using cysteine reagent. The results were analyzed statistically using ANOVA with Bonferroni post hoc tests. Results: Results showed a high significance in serum fucose in oral squamous cell carcinoma (OSCC) and leukoplakia subjects compared to normal controls. There was a gradual increase in the values noted from control to leukoplakia and to squamous cell carcinoma. Conclusions: Estimation of serum fucose may be a reliable marker and can be used as an effective diagnostic biomarker in oral squamous cell carcinoma patients.

Experimental Study on the Antiinflammatory Activities of Bojeasodok-um subtracted Scrophulariae Radix, Lasiosphaera seu Calvatia, Isatidis Radix added indigo Naturalis, Lithospermi Radix (보제소독음가감방(普濟消毒飮加減方)의 소염작용(消炎作用)에 관한 실험적(實驗的) 연구(硏究))

  • Kim, Sung-Hak;Park, Jong-Hyung;Kim, Kyung-Jum
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.23 no.3
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    • pp.1-10
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    • 2010
  • Objective : Erysipelas is an acute inflammation caused by pyogenic bacteria. This mainly involves the upper part of dermis. It begins as erythematous patches with tenderness, followed by fever, headache, chills and fatigue etc. It may results in edema, obstruction of lymphatics and sepsis. So this experiment is carried out for test whether the Bojeasodok-um subtracted Scrophulariae Radix, Lasiosphaera seu Calvatia, Isatidis Radix added indigo Naturalis, Lithospermi Radix have an anti-inflammatory effect and have suppression effect on immunocyte in the state of inflammation which induced by Erysipelas. Method : Experimental animals made use of 4-5 week-age(weight 20-25g) ICR(male) mouse. In the breeding farm, the lighting time was controlled from 7:00 am till 7:00 pm, the temperature was controlled So we concluded that BS is prospected as an anti-inflammatory agent to cure inflammation induced bywithin 18-23$23^{\circ}C$ and water and food were not limited.The freezing lyophilization powder which were extracted from Bojesodok-Um divided low dose group(200mg/kg-BSL) and high dose group(500mg/kg-BSH) and after melting in water, it was orally administered to the mouse. Compared with inflammation induced group which were induced by triggering-inflammation reagent Carageenan and Zymosan and normal contrast group, we measured the edema decrement effect,macrophage and spleen cell activation. Result : 1. BS has suppress inflammatory reaction induced by Carageenan. 2. BS has suppress increasing activation of abdominal cavity macrophage in the Carageenan and Zymosan induced inflammation. 3. BS has suppress increasing activation of spleen cell in the Carageenan and Zymosan induced inflammation. Based on the above result, BS was improved its suppression effect to the inflammatory reaction through the suppression of spleen cell and macrophage activation. So we concluded that BS is prospected as an anti-inflammatory agent to cure inflammation induced by Erysipelas.

Preconcentration, Separation and Determination of lead(II) with Methyl Thymol Blue Adsorbed on Activated Carbon Using Flame Atomic Absorption Spectrometry (불꽃원자 흡수 분광법으로 활성탄소에 흡착된 메틸티몰 블루로 납(II)의 예비농축, 분리 및 측정)

  • Ensafi, Ali A.;Ghaderi , Ali R.
    • Journal of the Korean Chemical Society
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    • v.52 no.1
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    • pp.16-22
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    • 2008
  • on-line system for preconcentration and separation of lead(II) is presented. The method is based on the complex formation of Pb(II) with adsorbed Methyl thymol blue on activated carbon. The conditions of preparing the solid phase reagent and of quantitative recovery of Pb(II) from diluted solutions, such as acidity of aqueous phase, solid phase capacity, and flow variables were studied as well as effect of potential interfering ions. After preconcentration step, the metal ions are eluted automatically by 5 ml of 0.5 M HNO3 solution and the lead ions content was determined by flame atomic absorption spectrometry. Under the optimum conditions, the lead ions in aqueous samples were separated and preconcentrated about 1000-fold by the column. The detection limit was 0.001 g mL-1. Lead has been determined in river and tap water samples, with recovery of 98 to 102%.

Characterization of Synthetic Polymeric AI(III) Inorganic Coagulants for Water Treatment (상수처리용 합성 무기고분자 Al(III)계 응집제의 화학적특성)

  • Han Seung-Woo;Jung Chul-Woo;Kang Lim-Seok
    • Journal of Environmental Science International
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    • v.8 no.6
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    • pp.711-716
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    • 1999
  • This research explored the feasibility of preparing and utilizing a prefonned polymeric solution of Al(III) for coagulation in water treatment. Slow base(NaOH) injection into supersaturated aluminum chloride and aluminum sulfate solutions did produce high yields of Al polymers useful to water treatment applications. The method of characterization analysis was based on timed spectrophotometer with ferron as a color developing reagent. The hydrolytic Al species were divided into $monomeric(Al_a),\;polymeric(Al_b),\;and\;precipitate(Al_c)$ from the difference in reaction kinetics. The analysis of PACl's characteristics showed that the quantity of polymeric Al produced at value of$ r(OH_{added}/AI)=2.2$ was $83\%$ of the total aluminum in solution, as showing maximum contents and precipitated Al was dramatically increased when r was increased above 2.35. In addition, the characteristics of polyaluminum sulfate (PAS) showed that polymeric Al contained at r = 0.75 was $18\%$ of the total aluminum in solution. The synthesized PACI and PAS were stable during storing period, as indicating negligible aging effect. The effect of sulfate ion on PACI was dependent on the concentration of sulfate ion. That is, polymeric species decrease and precipitate species increase as sulfate ion concentration increased. It can be concluded that the sulfate cause the formation of $Al(OH)_{3(S)}$ at low pH. However, The effect of calcium ion was negligible for distribution of Al species.

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Determination of Boron by Ion Pair Liquid Chromatography with Chromotropic Acid (Chromotropic Acid를 착화제로 이용한 이온쌍 액체 크로마토그래피에 의한 붕소의 분리와 정량)

  • Yun, Young Ja;Yu, Gu Yong
    • Journal of the Korean Chemical Society
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    • v.39 no.4
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    • pp.288-293
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    • 1995
  • The separation and determination of boron with chromotropic acid (1,8-Dihydroxynaphthalene-3,6-disulfonic acid) as a complex agent has been studied using ion pair liquid chromatography. The use of tetrabutylammonium bromide added as an ion pair reagent to mobile phase (MeOH 61%, phosphate buffer 39% pH=8.5) allowed good separation of boron-chromotrophic acid complex anion and chromotrophic acid on poly(styrene-divinylbenzene) based reversed phase column (PRP-1, 15 $cm{\times}4.6$ mm i.d.). The complex formation between boric acid and chromotrophic acid was enhanced in the presence of 0.1 M tetrabutylammonium bromide, resulting in high sensitivity. The linear calibration was achieved over the boron concentration range of 0.5∼1000 ${\mu}g/L.$ The detection limit was 0.5 ${\mu}g/L$ (S/N=2). The proposed method was applied to the determination of boron in commercially available chemicals, $Na_2SO_4$, NaOH, KCl.

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